Patent Grant
11954861
Devices and methods for collecting data for diagnostic purposes are disclosed. In particular, the devices and methods of the present application may be suitable for evaluating and tracking bacterial load in a wound over time.
Wound care is a major clinical challenge. Healing and chronic non-healing wounds are associated with a number of biological tissue changes including inflammation, proliferation, remodeling of connective tissues and, a common major concern, bacterial infection. A proportion of wound infections are not clinically apparent and contribute to the growing economic burden associated with wound care, especially in aging populations. Currently, the gold-standard wound assessment includes direct visual inspection of the wound site under white light combined with indiscriminate collection of bacterial swabs and tissue biopsies resulting in delayed, costly and often insensitive bacteriological results. This may affect the timing and effectiveness of treatment. Qualitative and subjective visual assessment only provides a gross view of the wound site, but does not provide information about underlying biological and molecular changes that are occurring at the tissue and cellular level. A relatively simple and complementary method that collects and analyzes ‘biological and molecular’ information in real-time to provide early identification of such occult change and guidance regarding treatment of the same is desirable in clinical wound management. Early recognition of high-risk wounds may guide therapeutic intervention and provide response monitoring over time, thus greatly reducing both morbidity and mortality due especially to chronic wounds.
In accordance with one aspect of the present disclosure, an adaptor for configuring a mobile communication device for tissue imaging comprises a housing, at least one excitation light source, a white light source, and a power source for powering the adaptor. The housing is configured to be removably coupled to a mobile communication device. The at least one excitation light source is configured to emit excitation light selected to elicit emission of bacterial autofluorescence data indicative of bacterial load of a tissue target in response to illumination of the tissue target by the excitation light. The white light source is configured to emit white light to elicit reflection data indicative of one or more of size, shape, volume, topography, depth, and area of the tissue target in response to illumination of the tissue target with the white light.
In accordance with another aspect of the present disclosure, an adaptor for configuring a mobile communication device for tissue imaging includes a housing, a first excitation light source, a second excitation light source, a thermal sensor, and a power source for powering the adaptor. The first excitation light source is configured to emit excitation light at a first wavelength or wavelength band selected to elicit emission of bacterial autofluorescence from a tissue target in response to illumination of the tissue target by the excitation light. The second excitation light source is configured to emit excitation light at a second wavelength or wavelength band selected to elicit emissions from one or more biomarkers associated with wound healing, wherein the first wavelength or wavelength band is different than the second wavelength or wavelength band. The thermal sensor is configured to detect thermal information emitted by the tissue target.
Additional objects and advantages of the disclosure will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the disclosure. The objects and advantages of the disclosure will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims.
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the disclosure, as claimed.
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the disclosure and together with the description, serve to explain the principles of the disclosure.
At least some features and advantages will be apparent from the following detailed description of embodiments consistent therewith, which description should be considered with reference to the accompanying drawings, wherein:
Although the following detailed description makes reference to illustrative embodiments, many alternatives, modifications, and variations thereof will be apparent to those skilled in the art. Accordingly, it is intended that the claimed subject matter be viewed broadly.
Reference will now be made in detail to various embodiments, examples of which are illustrated in the accompanying drawings. The various exemplary embodiments are not intended to limit the disclosure. To the contrary, the disclosure is intended to cover alternatives, modifications, and equivalents.
Conventional clinical assessment methods of acute and chronic wounds continue to be suboptimal. Such assessment methods usually are based on a complete patient history, qualitative and subjective clinical assessment with simple visual appraisal using ambient white light and the ‘naked eye,’ and can sometimes involve the use of color photography to capture the general appearance of a wound under white light illumination. Regular re-assessment of progress toward healing and appropriate modification of the intervention is also necessary. Wound assessment terminology is non-uniform, many questions surrounding wound assessment remain unanswered, agreement has yet to be reached on the key wound parameters to measure in clinical practice, and the accuracy and reliability of available wound assessment techniques vary.
Visual assessment is frequently combined with swabbing and/or tissue biopsies for bacteriological culture for diagnosis. Bacterial swabs are collected at the time of wound examination and have the noted advantage of providing identification of specific bacterial/microbial species. However, multiple swabs and/or biopsies often are collected randomly from the wound site, and some swabbing techniques may in fact spread the microorganisms around with the wound during the collection process thus affecting patient healing time and morbidity. This may be a problem especially with large chronic (non-healing) wounds where the detection yield for bacterial presence using current swabbing and biopsy protocols is suboptimal (diagnostically insensitive), despite many swabs being collected.
Thus, current methods for obtaining swabs or tissue biopsies from the wound site for subsequent bacteriological culture are based on a non-targeted or ‘blind’ swabbing or punch biopsy approach, and have not been optimized to minimize trauma to the wound or to maximize the diagnostic yield of the bacteriology tests. In addition, bacteriological culture results often take about 2-3 days to come back from the laboratory and can be inconclusive, thus delaying accurate diagnosis and treatment. Thus, conventional methods of obtaining bacterial swabs do not necessarily provide relevant data regarding the wound and cannot provide real-time detection of infectious status of wounds. The lack of a non-invasive method to objectively and rapidly evaluate wound repair at a biological level (which may be at greater detail than simply appearance or morphology based), and to aid in targeting of the collection of swab and tissue biopsy samples for bacteriology is a major obstacle in clinical wound assessment and treatment. An alternative method is highly desirable.
As wounds (chronic and acute) heal, a number of key biological changes occur at the wound site at the tissue and cellular level. Wound healing involves a complex and dynamic interaction of biological processes divided into four overlapping phases—haemostasis, inflammation, cellular proliferation, and maturation or remodeling of connective tissues—which affect the pathophysiology of wound healing. A common major complication arising during the wound healing process, which can range from days to months, is infection caused by bacteria and other microorganisms. This can result in a serious impediment to the healing process and lead to significant complications. All wounds contain bacteria at levels ranging from contamination, through colonization, critical colonization to infection, and diagnosis of bacterial infection is based on clinical symptoms and signs (e.g., visual and odorous cues).
The most commonly used terms for wound infection have included wound contamination, wound colonisation, wound infection and, more recently, critical colonisation. Wound contamination refers to the presence of bacteria within a wound without any host reaction; wound colonisation refers to the presence of bacteria within the wound which do multiply or initiate a host reaction; and critical colonisation refers to multiplication of bacteria causing a delay in wound healing, usually associated with an exacerbation of pain not previously reported but still with no overt host reaction. Wound infection refers to the deposition and multiplication of bacteria in tissue with an associated host reaction. In practice the term ‘critical colonisation’ can be used to describe wounds that are considered to be moving from colonisation to local infection. The challenge within the clinical setting, however, is to ensure that this situation is quickly recognized with confidence and for the bacterial bioburden to be reduced as soon as possible, perhaps through the use of topical antimicrobials. Potential wound pathogens can be categorised into different groups, such as, bacteria, fungi, spores, protozoa and viruses depending on their structure and metabolic capabilities. Although viruses do not generally cause wound infections, bacteria can infect skin lesions formed during the course of certain viral diseases. Such infections can occur in several settings including in health-care settings (hospitals, clinics) and at home or chronic care facilities. The control of wound infections is increasingly complicated, yet treatment is not always guided by microbiological diagnosis. The diversity of micro-organisms and the high incidence of polymicrobic flora in most chronic and acute wounds give credence to the value of identifying one or more bacterial pathogens from wound cultures. The early recognition of causative agents of wound infections can assist wound care practitioners in taking appropriate measures. Furthermore, faulty collagen formation arises from increased bacterial burden and results in over-vascularized friable loose granulation tissue that usually leads to wound breakdown.
Accurate and clinically relevant wound assessment is an important clinical tool, but this process currently remains a substantial challenge. Current visual assessment in clinical practice only provides a gross view of the wound site (e.g., presence of purulent material and crusting). Current best clinical practice fails to adequately use the critically important objective information about underlying key biological changes that are occurring at the tissue and cellular level (e.g., contamination, colonization, infection, matrix remodeling, inflammation, bacterial/microbial infection, and necrosis) since such indices are i) not easily available at the time of the wound examination and ii) they are not currently integrated into the conventional wound management process. Direct visual assessment of wound health status using white light relies on detection of color and topographical/textural changes in and around the wound, and thus may be incapable and unreliable in detecting subtle changes in tissue remodeling. More importantly, direct visual assessment of wounds often fails to detect the presence of bacterial infection, since bacteria are occult under white light illumination. Infection is diagnosed clinically with microbiological tests used to identify organisms and their antibiotic susceptibility. Although the physical indications of bacterial infection can be readily observed in most wounds using white light (e.g., purulent exudate, crusting, swelling, erythema), this is often significantly delayed, and the patient is already at increased risk of morbidity (and other complications associated with infection) and mortality. Therefore, standard white light direct visualization fails to detect the early presence of the bacteria themselves or identify the types of bacteria within the wound.
Wound progression is currently monitored manually. The National Pressure Ulcer Advisory Panel (NPUAP) developed the Pressure Ulcer Scale for Healing (PUSH) tool that outlines a five-step method of characterizing pressure ulcers. This tool uses three parameters to determine a quantitative score that is then used to monitor the pressure ulcer over time. The qualitative parameters include wound dimensions, tissue type, and the amount of exudate or discharge, and thermal readings present after the dressing is removed. A wound can be further characterized by its odor and color. Such an assessment of wounds currently does not include critical biological and molecular information about the wound. Therefore, all descriptions of wounds are somewhat subjective and noted by hand by either the attending physician or the nurse.
What is desirable is a robust, cost-effective non-invasive and rapid imaging-based method or device for collecting wound data and providing an analysis in real-time. The data and analysis can be used to objectively assess wounds for changes at the biological, biochemical and cellular levels and to rapidly, sensitively and non-invasively detecting the earliest presence of bacteria/microorganisms within wounds. Such a method or device for detection of critical biological tissue changes in wounds may serve an adjunctive role with conventional clinical wound management methods in order to guide key clinico-pathological decisions in patient care. Such a device may be compact, portable and capable of real-time non-invasive and/or non-contact interrogation of wounds in a safe and convenient manner, which may allow it to fit seamlessly into routine wound management practice and user friendly to the clinician, nurse and wound specialist. This may also include use of this device in the home-care environment (including self-use by a patient), as well as in military battlefield environments. In addition, such an image-based device may provide an ability to monitor wound treatment response and healing in real-time by incorporating valuable ‘biologically-informed’ image-guidance into the clinical wound assessment process. This may ultimately lead to potential new diagnosis, treatment planning, treatment response monitoring and thus ‘adaptive’ intervention strategies which may permit enhancement of wound-healing response at the individual patient level. Precise identification of the systemic, local, and molecular factors underlying the wound healing problem in individual patients may allow better tailored treatment.
In accordance with the present teachings, methods of analysis for data collected from a wound are provided. For example, the collection of fluorescence image data appears to be promising for improving clinical wound assessment and management. When excited by short wavelength light (e.g., ultraviolet or short visible wavelengths), most endogenous biological components of tissues (e.g., connective tissues such collagen and elastins, metabolic co-enzymes, proteins, etc.) produce fluorescence of a longer wavelength, in the ultraviolet, visible, near-infrared and infrared wavelength ranges.
Tissue autofluorescence imaging provides a unique means of obtaining biologically relevant information of normal and diseased tissues in real-time, thus allowing differentiation between normal and diseased tissue states, as well as the volume of the diseased tissue. This is based, in part, on the inherently different light-tissue interactions (e.g., absorption and scattering of light) that occur at the bulk tissue and cellular levels, changes in the tissue morphology and alterations in the blood content of the tissues. In tissues, blood is a major light absorbing tissue component (i.e., a chromophore). This type of technology is suited for imaging disease in hollow organs (e.g., GI tract, oral cavity, lungs, bladder) or exposed tissue surfaces (e.g., skin). An autofluorescence imaging device in accordance with the present disclosure may collect wound data that provides/allows rapid, non-invasive and non-contact real-time analysis of wounds and their composition and components, to detect and exploit the rich biological information of the wound to improve clinical care and management.
A device in accordance with the present disclosure: 1) provides image-guidance for tissue sampling, detecting clinically-significant levels of pathogenic bacteria and wound infection otherwise overlooked by conventional sampling and 2) provides image-guidance for wound treatment, accelerating wound closure compared with conventional therapies and quantitatively tracking long-term changes in bacterial bioburden and distribution in wounds.
U.S. Pat. No. 9,042,967 B2 to DaCosta et al., entitled “Device and Method for Wound Imaging and Monitoring,” and issued on May 26, 2015, discloses at least some aspects of a device configured to collect data for objectively assessing wounds for changes at the biological, biochemical and cellular levels and for rapidly, sensitively and non-invasively detecting the earliest presence of bacteria/microorganisms within wounds. This patent claims priority to PCT Application No. PCT/CA2009/000680 filed on May 2009, and to U.S. Provisional Patent Application No. 61/054,780, filed on May 20, 2008. The entire content of each of these above-identified patents, patent applications, and patent application publications is incorporated herein by reference.
In accordance with one aspect of the present teachings, a handheld portable device to examine skin and wounds in real-time is provided. The device instantly detects, visualizes, and analyzes bacteria and tissue composition. The device is a compact, handheld, device for noncontact and noninvasive imaging. It captures both white light (WL) and autofluorescence (AF) signals produced by tissue components and bacteria without the use of contrast agents. Although capable of detecting AF signals without use of contrast agents, one of ordinary skill in the art will understand that the devices disclosed herein can be used with contrast agents if desired. In addition to white light and fluorescence, the device also may capture thermal data from the imaged area. The device may be further configured to analyze the white light, fluorescence, and thermal data, correlate such data, and provide an output based on the correlation of the data, such as, for example, an indication of wound status, wound healing, wound infection, bacterial load, or other diagnostic information upon which an intervention strategy may be based.
The device may be configured to create and/or display composite images including green AF, produced by endogenous connective tissues (e.g., collagen, elastin) in skin, and red AF, produced by endogenous porphyrins in clinically relevant bacteria such as Staphylococcus aureus. Siderophores/pyoverdins in other species such as Pseudomonas aeruginosa appear blue-green in color with in vivo AF imaging. The device may provide visualization of bacterial presence, types, distribution, amounts in and around a wound as well as key information surrounding tissue composition (collagen, tissue viability, blood oxygen saturation). For example, the device may provide imaging of collagen composition in and around skin in real-time (via AF imaging).
In accordance with various exemplary embodiments of the present disclosure, the device may be configured to accurately detect and measure bacterial load in wounds in real-time, guide treatment decisions, and track wound healing over the course of antibacterial treatment. Additionally, bioluminescence imaging (BLI) may be used to correlate absolute bacterial load with FL signals obtained using the handheld device. The device may produce a uniform illumination field on a target area to allow for imagining/quantification of bacteria, collagen, tissue viability, and oxygen saturation.
The device may produce high-quality, focused images. The device may include software that provides macro zoom correction, auto-focus, auto white balance, wide dynamic range, noise reduction, image stabilization, and FL image calibration. In some embodiments, the device operates at an ambient temperature between about 0-35° C.
The device may be independent and self-contained. It may interface with computers, printers and EMR systems.
In accordance with one exemplary embodiment of the present disclosure, the device is configured to image bacteria in real-time (via, for example, fluorescence imaging), permitting ready identification of bacteria types, their location, distribution and quantity in accepted units of measurement and allowing identification of and distinction between several different species of bacteria. For example, autofluorescence imaging may be used to visualize and differentiate Pseudomonas aeruginosa (which fluoresces a greenish-blue colour when excited by 405 nm light from the device) from other bacteria (e.g., Staphylococcus aureus) that predominantly fluoresce a red/orange colour under the same excitation wavelength. In one exemplary embodiment the device's camera sensor and built in fluorescence multiband pass emission filter produce fluorescence images of bacteria (in wounds or normal skin) and Pseudomonas aeruginosa appear greenish-blue in colour while other bacteria emit a red/orange colour. The device detects differences in the autofluorescence emission of different endogenous molecules (called fluorophores) between the different bacteria.
In accordance with another exemplary embodiment of the present disclosure, the device is configured to identify or provide an indication of tissue viability in real-time (via fluorescence imaging). For example, blood preferentially absorbs 405 nm light compared with other visible wavelengths. Tissues which are perfused by blood are considered viable and can be differentiated from devitalized (poorly perfused) tissues using fluorescence imaging. Using 405 nm light from a device in accordance with the present teachings to illuminate a wound, the device can be configured with a multiband pass emission filter to detect the amount of 405 nm light that is absorbed or reflected from the tissues. Viable tissue contains blood that highly absorbs 405 nm light resulting in an image with low levels of 405 nm light, whereas nonviable (or devitalized) tissues do not contain sufficient blood and 405 nm is less absorbed. Thus, in an image of a wound where viable and nonviable tissues are present, the user will recognize viable tissues (from nonviable tissues) based on the relative amount of 405 nm light in the image, the viable tissues appearing darker compared with the nonviable tissues. In addition, in the green fluorescence “channel” of the resultant image (of the wound), viable tissues will appear less green fluorescent compared with nonviable tissues because viable tissues will preferentially absorb more of the 405 nm excitation light due to more blood being present, compared with nonviable tissues. Thus, while both viable and nonviable tissues in a resultant image obtained by the device may contain similar amounts of green fluorescent connective tissues (i.e., collagens), viable tissue will have less 405 nm excitation light to stimulate the connective tissue autofluorescence than nonviable tissues. The result is that viable tissues will have less green connective tissue fluorescence than non-viable tissues in the same image. The user will appreciate this difference visually during imaging with the device.
In accordance with another aspect of the present disclosure, the device is configured to capture and generate images and videos that provide a map or other visual display of user selected parameters. Such maps or displays may correlate, overlay, co-register or otherwise coordinate data generated by the device based on input from one or more device sensors. Such sensors may include, for example, camera sensors configured to detect white light and/or fluorescent images and thermal sensors configured to detect heat signatures of a target. For example, the device may be configured to display color images, image maps, or other maps of user selected parameters such as, for example, bacteria location and/or biodistribution, collagen location, location and differentiation between live tissues and dead tissues, differentiation between bacterial species, location and extent of blood, bone, exudate, temperature and wound area/size. These maps or displays may be output by the device based on the received signals and may be produced on a single image with or without quantification displays. The user-selected parameters shown on the map may be correlated with one or more wound parameters, such as shape, size, topography, volume, depth, and area of the wound. For example, in accordance with one exemplary embodiment, it is possible to use a ‘pseudo-coloured’ display of the fluorescence images/videos of wounds to color-code bacteria fluorescence (one colour) and connective tissues (another colour) etc. This may be accomplished by, for example, using a pixel-by-pixel coloring based on the relative amount of 405 nm light in the Blue channel of the resultant RGB image, green connective tissue fluorescence in the Green channel, and red bacteria fluorescence in Red channel. Additionally and/or alternatively, this may be accomplished by displaying the number of pixels in a given image for each of the blue, green and red channels which would represent amount of blood in tissue, amount of connective tissues, and amount of bacteria, respectively.
In accordance with one aspect of the present disclosure, the device may be configured to create and output reports regarding the collected data. For example, in accordance with one exemplary embodiment, the device user can generate a wound status report, which may include, for example, date/time, patient ID, images, etc. The user can export or print images, to a selected network, computer, printer when connected to cradle, and/or via USB to computer. The reports may be generated by the handheld device, by exporting data to a computer for processing and generation of reports, or by a combination of the two. Further, such reports, or the data contained therein, may form the basis of recommended intervention or treatment strategies. Reports may include, for example, medical reports, digital reports, reports that encompass handwritten input from clinicians (e.g., via tablet input, etc.). The reports may include various types of data including, for example, the identification of wound parameters and the tracking of these parameters over time. For example, the reports may identify and track changes in wound size, wound shape, wound topography, wound volume, wound area, bacterial load of the wound, location of bacteria within the wound, presence of exposed bone, blood, connective and other tissues, wound temperature, location of the wound on the patient, number of wounds on the patient, date of wound examination, patient identification, medications administered to the patient, interventional strategies and therapies as administered and as changed over time in response to changing wound parameters, etc. For example, the device may generate a report that tracks a patient's wound and skin status changes, including for example, wound size and bacterial burden over time. Further, the data collected may be used to generate a database that provides clinical data regarding wound parameters and the efficacy of various wound intervention/treatment strategies. Additionally, the device may be configured to integrate collected data/images/videos into the reports and, alternatively or additionally, include such reports and data/images/videos into a patient's electronic medical record (EMR). This process may be wirelessly, via the use of transfer cables, and the system also may be configured to upload the reports automatically.
The device has a memory sufficient to store several images/videos. In addition to internal memory, the device may include a Micro SD card interface for additional storage and firmware development. The device can inform the user of low memory capacity. The device may also include a data safeguard that will prompt a user to export files in the case of low memory availability.
In accordance with one aspect of the present disclosure, a method and device for fluorescence-based imaging and monitoring is disclosed. One exemplary embodiment of the device is a portable optical digital imaging device. The device may utilize a combination of white light, tissue fluorescence and reflectance imaging, and thermal imaging, and may provide real-time wound imaging, assessment, recordation/documentation, monitoring and/or care management. The device may be handheld, compact and/or light-weight. This device and method may be suitable for monitoring of wounds in humans and animals.
The device may generally comprise: i) one or more excitation/illumination light sources and ii) a detector device (e.g., a digital imaging detector device), which may be combined with one or more optical emission filters, or spectral filtering mechanisms, and which may have a view/control screen (e.g., a touch-sensitive screen), image capture and zoom controls. The device may also have: iii) a wired and/or wireless data transfer port/module, iv) an electrical power source and power/control switches, and/or v) an enclosure, which may be compact and/or light weight, and which may have a mechanism for attachment of the detector device and/or a handle grip. The excitation/illumination light sources may be LED arrays emitting light at about 405 nm (e.g., +/−5 nm), and may be coupled with additional band-pass filters centered at about 405 nm to remove/minimize the side spectral bands of light from the LED array output so as not to cause light leakage into the imaging detector with its own optical filters. The digital imaging detector device may be a digital camera, for example having at least an ISO800 sensitivity, but more preferably an ISO3200 sensitivity, and may be combined with one or more optical emission filters, or other equally effective (e.g., miniaturized) mechanized spectral filtering mechanisms (e.g., acousto-optical tunable filter or liquid crystal tunable filter). The digital imaging detector device may have a touch-sensitive viewing and/or control screen, image capture and zoom controls. The enclosure may be an outer hard plastic or polymer shell, enclosing the digital imaging detector device, with buttons such that all necessary device controls may be accessed easily and manipulated by the user. Miniature heat sinks or small mechanical fans, or other heat dissipating devices may be embedded in the device to allow excess heat to be removed from the excitation light sources if required. The complete device, including all its embedded accessories and attachments, may be powered using standard AC/DC power and/or by rechargeable battery pack. As discussed further below, the battery pack may be recharged with a charging stand.
The complete device may also be attached or mounted to an external mechanical apparatus (e.g., tripod, or movable stand with pivoting arm) allowing mobility of the device within a clinical room with hands-free operation of the device. Alternatively, the device may be provided with a mobile frame such that it is portable. The device may be cleaned using moist gauze wet with water, while the handle may be cleansed with moist gauze wet with alcohol. Additional appropriate cleaning methods will be apparent to those of ordinary skill in the art. The device may include software allowing a user to control the device, including control of imaging parameters, visualization of images, storage of image data and user information, transfer of images and/or associated data, and/or relevant image analysis (e.g., diagnostic algorithms). The device may also include one or more buttons/switches allowing a user to switch between white light and fluorescent light imaging.
A schematic diagram of an example of the device is shown in
The target object 10 may be marked with a mark 11 to allow for multiple images to be taken of the object and then being co-registered for analysis. The mark 11 may involve, for example, the use of exogenous fluorescence dyes of different colours which may produce multiple distinct optical signals when illuminated by the light sources 5 and be detectable within the image of the object 10 and thus may permit orientation of multiple images (e.g., taken over time) of the same region of interest by co-registering the different colours and the distances between them. The digital image acquisition device 1 may include one or more of: an interface 12 for a head-mounted display; an interface 13 for an external printer; an interface 14 for a tablet computer, laptop computer, desk top computer or other computer device; an interface 15 for the device to permit wired or wireless transfer of imaging data to a remote site or another device; an interface 16 for a global positioning system (GPS) device; an interface 17 for a device allowing the use of extra memory; and an interface 18 for a microphone.
The device may include a power supply 19 such as an AC/DC power supply, a compact battery bank, or a rechargeable battery pack. Alternatively, the device may be adapted for connecting to an external power supply. The device may have a housing 20 that houses all the components in one entity. The housing 20 may be equipped with a means of securing any digital imaging device within it. The housing 20 may be designed to be handheld, compact, and/or portable. The housing 20 may be one or more enclosures. The housing 20 may be comprised of a rugged material so that the device is tough and resistant to inadvertent drops by a user. Additionally, the housing 20 may include covers for any external ports of the device.
In accordance with one exemplary embodiment, the device may be charged while it is stationed in a charging stand, such as charging stand 30 shown in
An example of a device for fluorescence-based monitoring in accordance with the present disclosure is described below. All examples are provided for the purpose of illustration only and are not intended to be limiting. Parameters such as wavelengths, dimensions, and incubation time described in the examples may be approximate and are provided as examples only.
In this example, the device uses two violet/blue light (e.g., 405 nm+/−10 nm emission, narrow emission spectrum) LED arrays (Opto Diode Corporation, Newbury Park, California), each situated on either side of the imaging detector assembly as the excitation or illumination light sources. These arrays have an output power of approximately 1 Watt each, emanating from a 2.5×2.5 cm2, with a 70-degree illuminating beam angle. The LED arrays may be used to illuminate the tissue surface from a distance of about 10 cm, which means that the total optical power density on the skin surface is about 0.08 W/cm2. At such low powers, there is no known potential harm to either the target wound or skin surface, or the eyes from the excitation light. However, it may be inadvisable to point the light directly at any individual's eyes during imaging procedures. It should also be noted that 405 nm light does not pose a risk to health according to international standards formulated by the International Electrotechnical Commission (IEC), as further detailed on the website:
The one or more light sources may be articulated (e.g., manually) to vary the illumination angle and spot size on the imaged surface, for example by using a built-in pivot, and are powered for example through an electrical connection to a wall outlet and/or a separate portable rechargeable battery pack. Excitation/illumination light may be produced by sources including, but not limited to, individual or multiple light-emitting diodes (LEDs) in any arrangement including in ring or array formats, wavelength-filtered light bulbs, or lasers. Selected single and multiple excitation/illumination light sources with specific wavelength characteristics in the ultraviolet (UV), visible (VIS), far-red, near infrared (NIR) and infrared (IR) ranges may also be used, and may be composed of a LED array, organic LED, laser diode, or filtered lights arranged in a variety of geometries. Excitation/illumination light sources may be ‘tuned’ to allow the light intensity emanating from the device to be adjusted while imaging. The light intensity may be variable. The LED arrays may be attached to individual cooling fans or heat sinks to dissipate heat produced during their operation. The LED arrays may emit narrow 405 nm light, which may be spectrally filtered using a commercially available band-pass filter (Chroma Technology Corp, Rockingham, VT, USA) to reduce potential ‘leakage’ of emitted light into the detector optics. When the device is held above a tissue surface (e.g., a wound) to be imaged, the illuminating light sources may shine a narrow-bandwidth or broad-bandwidth violet/blue wavelength or other wavelength or wavelength band of light onto the tissue/wound surface thereby producing a flat and homogeneous field within the region-of-interest. The light may also illuminate or excite the tissue down to a certain shallow depth. This excitation/illumination light interacts with the normal and diseased tissues and may cause an optical signal (e.g., absorption, fluorescence and/or reflectance) to be generated within the tissue.
By changing the excitation and emission wavelengths accordingly, the imaging device may interrogate tissue components (e.g., connective tissues and bacteria in a wound) at the surface and at certain depths within the tissue (e.g., a wound). For example, by changing from violet/blue (˜400-500 nm) to green (˜500-540 nm) wavelength light, excitation of deeper tissue/bacterial fluorescent sources may be achieved, for example in a wound. Similarly, by detecting longer wavelengths, fluorescence emission from tissue and/or bacterial sources deeper in the tissue may be detected at the tissue surface. For wound assessment, the ability to interrogate surface and/or sub-surface fluorescence may be useful, for example in detection and potential identification of bacterial contamination, colonization, critical colonization and/or infection, which may occur at the surface and often at depth within a wound (e.g., in chronic non-healing wounds). In one example, referring to
Example embodiments of the device are shown in
This light signal produced by the excitation/illumination light sources may be detected by the imaging device using optical filter(s) (e.g., those available from Chroma Technology Corp, Rockingham, VT, USA) that reject the excitation light but allow selected wavelengths of emitted light from the tissue to be detected, thus forming an image on the display. There is an optical filter holder attached to the enclosure frame in front of the digital camera lens which may accommodate one or more optical filters with different discrete spectral bandwidths, as shown in insets b) and c) of
These band-pass filters may be selected and aligned in front of the digital camera lens to selectively detect specific optical signals from the tissue/wound surface based on the wavelength of light desired. Spectral filtering of the detected optical signal (e.g., absorption, fluorescence, reflectance) may also be achieved, for example, using a liquid crystal tunable filter (LCTF), or an acousto-optic tunable filter (AOTF) which is a solid-state electronically tunable spectral band-pass filter. Spectral filtering may also involve the use of continuous variable filters, and/or manual band-pass optical filters. These devices may be placed in front of the imaging detector to produce multispectral, hyperspectral, and/or wavelength-selective imaging of tissues.
The device may be modified by using optical or variably oriented polarization filters (e.g., linear or circular combined with the use of optical wave plates) attached in a reasonable manner to the excitation/illumination light sources and the imaging detector device. In this way, the device may be used to image the tissue surface with polarized light illumination and non-polarized light detection or vice versa, or polarized light illumination and polarized light detection, with either white light reflectance and/or fluorescence imaging. This may permit imaging of wounds with minimized specular reflections (e.g., glare from white light imaging), as well as enable imaging of fluorescence polarization and/or anisotropy-dependent changes in connective tissues (e.g., collagens and elastin) within the wound and surrounding normal tissues. This may yield useful information about the spatial orientation and organization of connective tissue fibers associated with wound remodeling during healing.
All components of the imaging device may be integrated into a single structure, such as an ergonomically designed enclosed structure with a handle, allowing it to be comfortably held with one or both hands. The device may also be provided without any handle. The device may be light weight, portable, and may enable real-time digital imaging (e.g., still and/or video) of any target surface (for example, the skin and/or oral cavity, which is also accessible) using white light, fluorescence and/or reflectance imaging modes. The device may be scanned across the body surface for imaging by holding it at variable distances from the surface, and the device may be used in a lit environment/room to image white light reflectance/fluorescence. The device may also be used in a dim or dark environment/room to optimize the tissue fluorescence signals and to minimize background signals from room lights. The device may be used for direct (e.g., with the unaided eye) or indirect (e.g., via the viewing screen of the digital imaging device) visualization of wounds and surrounding normal tissues.
The device may also be embodied as not being handheld or portable, for example as being attached to a mounting mechanism (e.g., a tripod or stand) for use as a relatively stationary optical imaging device for white light, fluorescence and reflectance imaging of objects, materials, and surfaces (e.g., a body). This may allow the device to be used on a desk or table or for ‘assembly line’ imaging of objects, materials and surfaces. In some embodiments, the mounting mechanism may be mobile.
Other features of this device may include the capability of digital image and video recording, possibly with audio, methods for documentation (e.g., with image storage and analysis software), and wired or wireless data transmission for remote telemedicine/E-health needs.
In some embodiments, the image acquisition device may be a mobile device, such as a cellular telephone or smartphone. In these embodiments, the mobile device is used to obtain the white light and/or fluorescent images. As discussed further below, the image acquisition device may also include an adaptor for attachment to the mobile device. The insets e) and f) of
In order to demonstrate the capabilities of the image acquisition device in wound care and other relevant applications, a number of feasibility experiments were conducted using the particular example described above. It should be noted that all fluorescence imaging experiments used a Sony camera (Sony Cybershot DSC-T200 Digital Camera, Sony Corporation, North America) as described above. The camera settings were set so that images were captured without a flash, and with the ‘Macro’ imaging mode set. Images were captured at 8 megapixels. The flash was used to capture white light reflectance images. All images were stored on the xD memory card for subsequent transfer to a personal computer for long-term storage and image analysis.
In one exemplary embodiment, white light reflectance and fluorescence images/movies captured with the device were imported into Adobe Photoshop for image analysis. However, image analysis software was designed using MatLab™ (Mathworks) to allow a variety of image-based spectral algorithms (e.g., red-to-green fluorescence ratios, etc.) to be used to extract pertinent image data (e.g., spatial and spectral data) for quantitative detection/diagnostic value. Image post-processing also included mathematical manipulation of the images.
In accordance with another exemplary embodiment of the present disclosure, a handheld device for collection of data from a wound includes a low-cost, consumer-grade, Super HAD™ charge-coupled device (CCD) sensor-based camera (Model DSC-T900, Sony Corp., Japan), with a 35 to 140 mm equivalent 4× zoom lens housed in a plastic body and powered by rechargeable batteries. An exemplary embodiment of this handheld imaging device is shown in
Inset (b) of
The device of
In another exemplary embodiment, the image acquisition device is a handheld device that is incorporated with a mobile device to take both white light images and fluorescent images. It is also contemplated that in some embodiments, the handheld device takes only white light images or only fluorescent images when incorporated with the mobile device. The mobile device may be a mobile communication device, such as a smartphone, mobile phone, iPod, iPhone, or other such device having existing image-capturing capabilities such as the CCD sensor. Although described herein with regard to usage with the iPod touch or iPhone, it should be understood that other platforms (e.g., Android, etc.) may be used. For example, as shown in
A mobile imaging device adaptor 200 is shown in
Inset (a) of
White light imaging allows the user to capture an image of a patient wound, and the fluorescence imaging allows the user to capture a corresponding image highlighting the presence of bacteria on the image. Both white light images and fluorescence images are viewed on the display screen 260 of the mobile device. Thus, the display screen 260 may be coupled to a camera on the mobile device. The display screen 260 may range between about 4-inches (diagonal) and about 7-inches (diagonal) widescreen display with Multi-Touch IPS technology. Other size displays may be used based on user needs. In one example, the display quality settings are 1136×640-pixel resolution at 326 pixels per inch; 800:1 contrast ratio; and 500 cd/m2 max brightness. The display may have a fingerprint-resistant oleophobic coating. The resolution of the camera may be about 5 Megapixels and may have resolutions higher than 5 Megapixels, such as up to about 24 Megapixels, depending upon availability, amount of storage available, etc. The selection of the lens design allows the production of high-quality images, specifically in the red and green spectra. In one exemplary embodiment, a five-element lens is used (as iPod touch design). The user can tap to focus video and/or still images. The camera has optimal performance in the dark. The camera has an LED flash and shutter speeds are high.
As shown in
The adaptor includes a fluorescent light source as discussed above. The fluorescent light source may be a violet light source that may emit excitation light in the range of about 400 nm-about 450 nm. It is also contemplated that the fluorescent light source emits excitation light in the range of about 450 nm-about 500 nm, about 500 nm-about 550 nm, about 600 nm-about 650 nm, about 650 nm-about 700 nm, about 700 nm-about 750 nm, and combinations thereof.
In accordance with this exemplary embodiment, the housing 205 of the adaptor 200 may be made by 3D printing. Other types of suitable structures are disclosed herein, and variations thereof will be understood by those of ordinary skill in the art based on the present teachings. The housing 205 provides a means for aligning the optical components with a consumer grade camera and encasing both the electrical components used to drive the LED and the thermal solution while creating a user friendly and lightweight handheld design. As shown in
As shown in
To perform fluorescence imaging using the adaptor 200, 300, the user switches on the violet LED using the toggle switch on the back of the device (
Those of ordinary skill in the art will understand that the adaptors described and illustrated herein are exemplary only, and that various other types and/or configurations of adaptors are contemplated without departing from the scope of the present disclosure and claims.
The studies discussed herein aimed to determine the ability of the handheld device to accurately detect and measure bacterial wounds in real time, guide treatment decisions, and track wound healing over the course of antibacterial treatment.
In one example, a study using the handheld device described herein tracked patient wounds over time. The study was broken into two parts, the first part to establish the safety and feasibility of AF imaging to improve wound sampling by accurately detecting clinically-significant levels of pathogenic bacteria in chronic wound patients, compared to standard wound assessment (including swab-based methods). The second part to demonstrate the feasibility of AF image guidance for wound treatment and quantitative treatment response, compared to standard wound assessment (including swab-based methods). Swab cultures were used to compare AF imagining with WL examination, to determine sensitivity, specificity, and predictive values for FL imaging for detecting clinically-significant bacterial loads.
In the first part of the study, high resolution WL and FL images were taken of every patient's wound at each visit. A disposable length calibration scale (sticker) was placed near the wound during WL and FL imaging to track each patient's ID and date of the imaging. Regular room lighting was used during WL imaging, and the lights were turned off during FL imaging to eliminate any artifacts in the images. To preserve bacterial characteristics on the tissue, no swabs were taken of the wound until completion of both the WL and FL imaging. The process to capture a WL image took 1-2 min per wound, and subsequent FL imaging took 1-2 min per wound. The clinician also swabbed each suspicious marked area on the patient using the Levine sampling method, and swabs were sent for blinded microbiology testing (it is noted that the Levine sampling method is the most commonly used swabbing method and involves only sampling the center of the wound). Patients were treated and discharged according to standard protocols.
The location(s) of red and/or green AF were marked on printed images, as discussed further below. FL spectroscopy was used in some cases to characterize AF areas in/around the wound. Spectra were compared on a location basis with microbiology results. A complete data file for each patient's visit (CSS, WL and FL images, spectroscopy and microbiology) were stored in an electronic database according to Good Clinical Practice guidelines.
In the second part of the study, three sequential 2-month arms were used: non-guided treatment (control), FL guided treatment and non-guided treatment (control). In the first 2-month phase, wounds were assessed weekly by CSS and then treated at the discretion of the clinical team using best practice methods (ultrasonic and/or scalpel wound debridement, topical/systemic antibiotics, saline wash, dry or anti-microbial dressings or iodine). Corresponding WL and FL images were taken of each wound pre- and post-treatment as described previously. 2-month evaluation periods were selected based on established clinical data for venous leg ulcers showing that this is sufficient to detect a reliable and meaningful change in wound area, as a predictive indicator of healing. Wound swabs were collected by FL guidance. Clinicians were blinded to FL images during this first (control) phase. During the subsequent 2-month phase, wound assessment was performed normally but clinicians were shown FL images of the wound during treatment.
During the final 2-month phase, WL and FL imaging were performed and swabs were collected, with clinician blinding to the FL results during treatment delivery. Importantly, while the clinicians understood and could remember the meaning and characteristics of the red and green fluorescence signals, respectively, blinding them during treatment delivery in the control periods was possible because the fluorescence results for each wound examination and each patient were different. Thus, in the absence of real-time fluorescence guidance during wound treatment, previous knowledge of fluorescence characteristics did not substantively influence the treatment decisions during the control periods. WL and FL images were also taken after each treatment to analyze wound area.
Four blinded, trained clinical and/or research staff members independently measured the average wound size on WL images using digital tracing (MATLAB v.7.9.0, The MathWorks, Massachusetts, USA). The observers measured the wounds in separate sessions with a minimum of 7 days between sessions to minimize memory effect. An adhesive scale bar placed adjacent to the wound during imaging provided accurate length calibration within +0.5 mm. Room lights remained on during WL imaging, but were turned off during FL imaging. WL and FL images were collected with the handheld device held/positioned 10-15 cm from the wound. All imaging parameters (distance, exposure time, ISO setting, white balance, etc.) were kept constant between visits. For distances less than 5 cm from a wound (small diameter wounds), the camera's built-in macro mode was used. Automatic focusing allowed rapid (˜1 s) image acquisition. Images (or video) were captured in real-time and stored on the camera's memory card. Switching between WL and FL modes was substantially instantaneous using a built in “toggle switch.” Devices were decontaminated between uses with 70% ethyl alcohol.
WL and AF images were transferred to a laptop. Regions of interest (ROIs) were identified from individual 1024×1024 pixel FL images of each wound at each clinic visit. RGB images were separated into individual channels. The green and red channels of the RGB image were representative of the true tissue and bacterial AF signals detected in vivo. To quantify bacterial levels from individual FL images, the following image processing procedures were used. Briefly, individual green and red image channels from each RGB image were converted to greyscale (the blue channel was not used) and pixels whose greyscale intensity was above a given histogram threshold (selected to reduce the background noise of the raw image) were counted. A red color mask for red FL bacteria was created by finding the local maxima in the color range 100-255 greyscale. Then, an inverted green color mask was used to remove the green FL. All pixels with red FL (above the histogram threshold) were binarized and the sum of all “1” pixels was calculated. This was repeated for the green channel of each image. These data gave an estimate of the amount of red (or green) bacteria in each image. The number of FL pixels was converted into a more useful pixel area measure (cm2) using the adhesive length calibration stickers, thereby providing the total amount of fluorescent bacteria as an area measurement. The Levine method was used to aseptically swab wounds for confirmation of bacterial presence, species typing, Gram signing, antibiograms, and semi-quantitative bacterial load.
Tissue AF produced by endogenous collagen or elastin in the skin appeared as green FL, and clinically-relevant bacterial colonies (e.g. Staphylococcus aureus) appeared as red FL (caused by endogenous porphyrins). Some bacteria (e.g. Pseudomonus aeruginosa) produced a blue-green signal, due to siderophores/pyoverdins, which was differentiated spectrally and texturally from dermis AF using image analysis software. WL and FL images were collected in less than 1 second by the high-sensitivity CCD sensor mounted with a dual band FL filter (λemiss=500-550 and 590-690 nm) (Chroma Technologies Corp, VT, USA). The CCD image sensor was sensitive across a broad wavelength range of ˜300-800 nm. The handheld device integrated easily into the routine clinical work flow. By combining tissue FL with bacterial FL in a single composite image, the clinician instantly visualized the distribution and extent of the bacterial load within the anatomical context of the wound and body site. Typically, FL imaging added approximately 1-3 minutes/patient to the wound assessment routine, depending on the number of wounds and the duration of FL-guided swabbing.
The variation in measurements of wound areas between images taken under WL and FL were compared. The correlation between change in average wound area and FL image-guided treatment using Pearson correlation coefficients was also calculated. Assessing changes in wound area between the first control, the second FL image-guided, and the third control periods were performed using a linear mixed effect model.
The accuracy of identifying clinically-significant bacterial load for AF image-guided was compared with swabbing techniques and WL imaging. A total of 490 swabs were collected, of which 36.9% were taken from wound beds, 30.2% from wound peripheries, and 32.9% from “off-site” areas. It was determined that the AF accurately determined 74.5% of wounds with clinically-significant bioburden, and that WL imaging only detected 52.5% of the wounds. The handheld device accurately determined clinically-significant bioburden 82.4% of the time in the wound periphery and 67.1% in other areas. WL examination was correct only 17.6% of the time in peripheries and 32.9% in other areas. The overall accuracy of judging the presence of clinically-significant bacterial in chronic wounds for AF was 74.5% versus 35.5% for traditional methods of WL and swab results.
AF imaging detected clinically significant bacterial load in 85% of wound peripheries missed by conventional methods. Thus, the Levine method for swabbing only the wound bed may be insufficient, possibly resulting in antibacterial treatment being inappropriately withheld. However, modifying standard sampling practices to include swabbing of the wound periphery of all wounds would be impractical and costly. AF imaging could help clinicians decide if and where wound margins require sampling. The handheld imaging device also identified clinically significant bioburden in surrounding locations close to wounds, which represent sites of potential re-infection, where traditional methods do not examine or swab.
Identifying and quantitating wound bacterial burden is an important determinant of infection and healing. Data on the visualization and quantitative tracking of bacterial load led to the identification of a new, simple method for image-guided debridement and topical application of antibiotic and antiseptic, which minimizes unnecessary trauma to the wound boundary and maximizes the contribution of debridement to reducing bacterial burden. Every wound has the potential for infection, but distinguishing true infection from critical colonization by best practice methods remains challenging and arbitrary, and can lead to over- and under-treatment.
The handheld imaging device identifies pathogenic microbes and differentiates between at least the two major pathogenic species (P. aeruginosa and S. aureus), and informs medical treatment decisions. The device also offers a quantitative and reproducible way to monitor the effectiveness of existing and emerging wound care treatments. Furthermore, the handheld imaging device can be used to diagnose critically colonized wounds.
Multiple variables including host response, local and systemic factors, malperfusion, immunosuppression, diabetes, and medications affect the risk of infection. Critically colonized wounds can be difficult to diagnose because they do not always display classical signs of infection or clearly elevated levels of bioburden. Indeed, the clinical relevance of differentiating critically colonized wounds from infected wounds remains controversial. Identifying a high bacterial load in asymptomatic patients before infection occurs using AF imaging may help prevent infections by prompting aggressive treatment. If a bacterial infection is suspected, antibiotic selection could be guided by the established clinical principles and by AF identification of heavy bacterial burden and differentiation between Gram negative P. aeruginosa and Gram positive S. aureus.
In another exemplary embodiment, image analysis may be carried out on the handheld device or WL and FL images may be transferred to a laptop for image processing. Image analysis and processing of image data may be performed using a processor of the handheld device, and the results of such analyses may be displayed on the display of the handheld device.
The following two exemplary programs may be used for image processing (for example, analysis of the data collected by the exemplary device using the Super HAD™ charge-coupled device (CCD) sensor-based camera (Model DSC-T900)) and portions of these processes are illustrated in
A red color mask for red FL bacteria is created by finding the local maxima in the color range 100 to 255 gray scale. Then, an inverted green color mask is used to remove the green FL. All pixels with red FL (above the histogram threshold) are binarized and the sum of all “1” pixels is calculated. This is repeated for the green channel of each image. These data give an estimate of the amount of red bacteria in each image. The number of FL pixels is converted into a more useful pixel area measure (cm2) by applying a ruler on the pixel image, thereby providing the total amount of fluorescent bacteria as an area measurement (cm2). The sizes of the wounds may be traced and measured similarly by converting pixel areas to cm2 of the circled wound area on the WL images. The resolution of the FL images is sufficient to localize bacteria based on regions of FL. ImageJ software may be used to separate FL images into red, green, and blue channels using the built-in batch processing function “Split Channels” located within the image menu and color submenu of the camera. Each resulting channel is displayed and saved in gray scale. For further analysis, an ROI may be identified in each corresponding red, green, and blue channel image. Under the built-in analysis menu, the “Set Measurement” function may be used to select and measure the following measurement parameters for each color channel image: pixel area, min. and max. gray scale intensity values, and mean gray intensity values. The average red channel intensity value may be determined as (bacterial) FL intensity per square pixel in each red channel image and then used for data analysis and comparison.
In one exemplary embodiment, a mouse skin wound model was used to correlate wound status with the progression of bacterial infection (n=5; 8 to 12 weeks; NCRNU-F). Correlation was based on data obtained using the exemplary handheld device described above, which incorporates the Super HAD™ charge-coupled device (CCD) sensor-based camera (Model DSC-T900. Daily WL and FL images were taken of the wounds as they became infected over time. Antibacterial treatment (topical Mupirocin three times daily, for a total of 1 day) was applied to the wound site when the red FL intensity peaked. The anti-microbial effect of the treatment was monitored over time using the handheld device to acquire daily WL and FL images of the wound after treatment. The wounds were monitored for a total of 10 days (see
In accordance with another exemplary embodiment, BLI can be used to measure the absolute amount of bacteria in vivo, because it is one of the most sensitive and reliable screening tools for determining bacterial load. BLI collects the light emitted from the enzymatic reaction of luciferase and luciferin and therefore does not require excitation light. FL imaging using the handheld device (without any exogenous FL contrast agent administration) and BLI imaging of inoculated S. aureus bacteria were tracked over time and the FL and BLI intensities were compared (see
Imaging of Bacteriological Samples
Imaging devices in accordance with the present disclosure may be useful for imaging and/or monitoring in clinical microbiology laboratories. Such devices may be used for quantitative imaging of bacterial colonies and quantifying colony growth in common microbiology assays. Fluorescence imaging of bacterial colonies may be used to determine growth kinetics. Software may be used to provide automatic counting of bacterial colonies.
To demonstration the utility of such devices in a bacteriology/culture laboratory, live bacterial cultures were grown on sheep's blood agar plates. Bacterial species included Streptococcus pyogenes, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa (American Type Culture Collection, ATCC). These were grown and maintained under standard incubation conditions at 37° C. and used for experimentation when during ‘exponential growth phase’. Once colonies were detected in the plates (˜24 h after inoculation), the device was used to image agar plates containing individual bacterial species in a darkened room. Using violet/blue (about 405 nm) excitation light, the device was used to image both combined green and red autofluorescence (about 490-550 nm and about 610-640 nm emission) and only red autofluorescence (about 635+/−10 nm, the peak emission wavelength for fluorescent endogenous porphyrins) of each agar plate. Fluorescence images were taken of each bacterial species over time for comparison and to monitor colony growth.
Reference is now made to
High spatial resolution of the camera detector combined with significant bacterial autofluorescence signal-to-noise imaging with the device allowed detection of very small (e.g., <1 mm diameter) colonies. The device provided a portable and sensitive means of imaging individual bacterial colonies growing in standard agar plates. This provided a means to quantify and monitor bacterial colony growth kinetics, as seen in inset c), as well as potentially monitoring response to therapeutic intervention, with antibiotics or photodynamic therapy (PDT) as examples, over time using fluorescence. Therefore, devices in accordance with the present disclosure may serve as a useful tool in the microbiology laboratory.
In addition to providing detecting of bacterial species, the device may be used for differentiating the presence and/or location of different bacterial species (e.g., Staphylococcus aureus or Pseudomonas aeruginosa), for example in wounds and surrounding tissues. This may be based on the different autofluorescence emission signatures of different bacterial species, including those within the 490-550 nm and 610-640 nm emission wavelength bands when excited by violet/blue light, such as light around 405 nm. Other combinations of wavelengths may be used to distinguish between other species on the images. This information may be used to select appropriate treatment, such as choice of antibiotic.
Such imaging of bacteriology samples may be applicable to monitoring of wound care.
Use in Monitoring of Wound Healing
Devices in accordance with the present disclosure may also be scanned above any wound (e.g., on the body surface) such that the excitation light may illuminate the wound area. The wound may then be inspected using the device such that the operator may view the wound in real-time, for example, via a viewer on the imaging device or via an external display device (e.g., heads-up display, a television display, a computer monitor, LCD projector or a head-mounted display). It may also be possible to transmit the images obtained from the device in real-time (e.g., via wireless communication) to a remote viewing site, for example for telemedicine purposes, or send the images directly to a printer or a computer memory storage. Imaging may be performed within the routine clinical assessment of patient with a wound.
Prior to imaging, fiduciary markers (e.g., using an indelible fluorescent ink pen) may be placed on the surface of the skin near the wound edges or perimeter. For example, four spots, each of a different fluorescent ink color from separate indelible fluorescent ink pens, which may be provided as a kit to the clinical operator, may be placed near the wound margin or boundary on the normal skin surface. These colors may be imaged by the device using the excitation light and a multispectral band filter that matches the emission wavelength of the four ink spots. Image analysis may then be performed, by co-registering the fiduciary markers for inter-image alignment. Thus, the user may not have to align the imaging device between different imaging sessions. This technique may facilitate longitudinal (i.e., over time) imaging of wounds, and the clinical operator may therefore be able to image a wound over time without the need for aligning the imaging device during every image acquisition.
In addition, to aid in intensity calibration of the fluorescence images, a disposable simple fluorescent standard ‘strip’ may be placed into the field of view during wound imaging (e.g., by using a mild adhesive that sticks the strip to the skin temporarily). The strip may be impregnated with one or several different fluorescent dyes of varying concentrations which may produce pre-determined and calibrated fluorescence intensities when illuminated by the excitation light source, which may have single (e.g., 405 nm) or multiple fluorescence emission wavelengths or wavelength bands for image intensity calibration. The disposable strip may also have the four spots as described above (e.g., each of different diameters or sizes and each of a different fluorescent ink color with a unique black dot placed next to it) from separate indelible fluorescent ink pens. With the strip placed near the wound margin or boundary on the normal skin surface, the device may be used to take white light and fluorescence images. The strip may offer a convenient way to take multiple images over time of a given wound and then align the images using image analysis. Also, the fluorescent ‘intensity calibration’ strip may also contain an added linear measuring apparatus, such as a ruler of fixed length to aid in spatial distance measurements of the wounds. Such a strip may be an example of a calibration target which may be used with the device to aid in calibration or measuring of image parameters (e.g., wound size, fluorescence intensity, etc.), and other similar calibration target may be used.
It may be desirable to increase the consistency of imaging results and to reproduce the distance between the device and the wound surface, since tissue fluorescence intensity may vary slightly if the distance changes during multiple imaging sessions. Therefore, in an embodiment, the device may have two light sources, such as low power laser beams, which may be used to triangulate individual beams onto the surface of the skin in order to determine a fixed or variable distance between the device and the wound surface. This may be done using a simply geometric arrangement between the laser light sources, and this may permit the clinical operator to easily visualize the laser targeting spots on the skin surface and adjust the distance of the device from the wound during multiple imaging sessions. Other methods of maintaining a constant distance may include the use of ultrasound, or the use of a physical measure, such as a ruler, or a range finder mechanism.
Use in White Light Imaging
Devices in accordance with the present disclosure may also be used to take white light images of the total wound with surrounding normal tissues using a measuring apparatus (e.g., a ruler) placed within the imaging field of view. This may allow visual assessment of the wound and calculation/determination of quantitative parameters such as the wound area, circumference, diameter, and topographic profile. Wound healing may be assessed by planimetric measurements of the wound area at multiple time points (e.g., at clinical visits) until wound healing. The time course of wound healing may be compared to the expected healing time calculated by the multiple time point measurements of wound radius reduction using the equation R=√A/π (R, radius; A, planimetric wound area; π, constant 3.14). This quantitative information about the wound may be used to track and monitor changes in the wound appearance over time, in order to evaluate and determine the degree of wound healing caused by natural means or by any therapeutic intervention. This data may be stored electronically in the health record of the patient for future reference. White light imaging may be performed during the initial clinical assessment of the patient by the operator.
Use in Autofluorescence Imaging
Devices in accordance with the present disclosure may also be designed to detect all or a majority of tissue autofluorescence (AF). For example, using a multi-spectral band filter, the device may image tissue autofluorescence emanating from the following tissue biomolecules, as well as blood-associated optical absorption, for example under 405 nm excitation: collagen (Types I, II, III, IV, V and others) which appear green, elastin which appears greenish-yellow-orange, reduced nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), which emit a blue-green autofluorescence signal, and bacteria/microorganisms, most of which appear to have a broad (e.g., green and red) autofluorescence emission.
Image analysis may include calculating a ratio of red-to-green AF in the image. Intensity calculations may be obtained from regions of interest within the wound images. Pseudo-coloured images may be mapped onto the white light images of the wound.
Reference is now made to
To test the ability of the device to detect connective tissues and several common bacteria present in typical wounds, a sample of pig meat with simulated wounds was prepared by applying six bacterial species to each of six small 1.5 cm2 wound incision sites on the skin surface: Streptococcus pyogenes, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa. An additional small incision was made in the meat skin, where no bacteria were added, to serve as a control. However, it was expected that bacteria from the other six incisions sites would perhaps contaminate this site in time. The device was used to image the bacteria-laden meat sample using white light reflectance and violet/blue light-induced tissue autofluorescence emission, using both a dual emission band (450-505 nm and 590-650 nm) emission filter and a single band (635+/−10 nm) emission filter, on the left and a single band filter over the course of three days, at 24 h time intervals, during which the meat sample was maintained at 37° C. Imaging was also performed on the styrofoam container on which the meat sample was stored during the three days.
The device mapped biodistribution of bacteria within the wound site and on the surrounding skin and thus may aid in targeting specific tissue areas requiring swabbing or biopsy for microbiological testing. Furthermore, using the imaging device may permit the monitoring of the response of the bacterially-infected tissues to a variety of medical treatments, including the use of antibiotics and other therapies, such as antibiotics, wound debridement, wound cleaning, photodynamic therapy (PDT), hyperbaric oxygen therapy (HOT), low level light therapy, or anti-matrix metalloproteinase (MMP). The device may be useful for visualization of bacterial biodistribution at the surface as well as within the tissue depth of the wound, and also for surrounding normal tissues. The device may thus be useful for indicating the spatial distribution of an infection.
Reference is now made to
The handheld device in accordance with the present disclosure spectrally distinguishes bacteria from connective tissues and blood in vivo. Using λexc=405_20 nm and λemiss=500 to 550 nm, 590 to 690 nm, the device detects AF signals of S. aureus, Staphylococcus epidermidis, P. aeruginosa, Candida, Serratia marcescens, Viridans streptococci (α-hemolytic streptococci), Streptococcus pyogenes (β-hemolytic streptococci), Corynebacterium diphtheriae, Enterobacter, Enterococcus, and methicillin-resistant S. aureus (MRSA), as verified by microbiological swab cultures (data from a human clinical trial by our group to be published in a forthcoming paper). This is a representative of the major types of pathogenic bacteria commonly found in infected wounds. Clinical microbiology tests confirmed that S. aureus, S. epidermidis, Candida, S. marcescens, Viridans streptococci, Corynebacterium diphtheriae, S. pyogenes, Enterobacter, and Enterococcus produced red FL (from porphyrin) while P. aeruginosa produced a bluish-green FL (from pyoverdin) detected by the handheld device. These spectral characteristics differ significantly from connective tissues (collagen, elastin) and blood, which appear green and dark red, respectively. A representative image of these spectral characteristics is shown in
The device was also able to detect spreading of the bacteria over the surface of the meat over time. Inset d) of
This example demonstrates the use of devices in accordance with the present disclosure for real-time detection of biological changes in connective tissue and bacterial growth based on autofluorescence alone, suggesting a practical capability of the device to provide longitudinal monitoring of bacterial growth in wounds.
Referring again to
In addition to detection of bacteria in wounds and on the skin surface, the device was also able to identify suspicious areas of muscle tissue, which may then be interrogated further by surgery or targeted biopsy for pathological verification, or by other optical means such as fluorescence spectroscopy using a fiber optic probe. Also, it detected contamination by various bacteria on the surface of the Styrofoam container on which the meat sample was kept. Autofluorescence of the bacteria appears as streaks of green and red fluorescence under violet/blue excitation light from the various bacterial species previously applied to the meat.
In order to determine the autofluorescence characteristics of bacteria growing in culture and in simulated skin wounds, hyperspectral/multispectral fluorescence imaging was used to quantitatively measure the fluorescence intensity spectra from the bacteria under violet/blue light excitation. Reference is now made to
This example shows that bacteria emit green and red autofluorescence, with some species (e.g., Pseudomonas aeruginosa) producing more of the former. Escherichia coli produced significant red autofluorescence from endogenous porphyrins. Such intrinsic spectral differences between bacterial species are significant because it may provide a means of differentiating between different bacterial species using autofluorescence alone. Excitation-emission matrices (EEMs) were also measured for each of the bacterial species used in these pilot studies, which confirmed that under violet/blue light excitation, all species produced significant green and/or red fluorescence, the latter being produced by porphyrins. Spectral information derived from excitation-emission matrices may aid in optimizing the selection of excitation and emission wavelength bandwidths for use with optical filters in the imaging device to permit inter-bacterial species differentiating ex vivo and in vivo. In this way, the device may be used to detect subtle changes in the presence and amount of endogenous connective tissues (e.g. collagens and elastins) as well as bacteria and/or other microorganisms, such as yeast, fungus and mold within wounds and surrounding normal tissues, based on unique autofluorescence signatures of these biological components.
This device may be used as an imaging and/or monitoring device in clinical microbiology laboratories. For example, the device may be used for quantitative imaging of bacterial colonies and quantifying colony growth in common microbiology assays. Fluorescence imaging of bacterial colonies may be used to determine growth kinetics.
Imaging of Blood in Wounds
Angiogenesis, the growth of new blood vessels, is an important natural process required for healing wounds and for restoring blood flow to tissues after injury or insult. Angiogenesis therapies, which are designed to “turn on” new capillary growth, are revolutionizing medicine by providing a unified approach for treating crippling and life-threatening conditions. Angiogenesis is a physiological process required for wound healing. Immediately following injury, angiogenesis is initiated by multiple molecular signals, including hemostatic factors, inflammation, cytokine growth factors, and cell-matrix interactions. New capillaries proliferate via a cascade of biological events to form granulation tissue in the wound bed. This process may be sustained until the terminal stages of healing, when angiogenesis is halted by diminished levels of growth factors, resolution of inflammation, stabilized tissue matrix, and endogenous inhibitors of angiogenesis. Defects in the angiogenesis pathway impair granulation and delay healing, and these are evident in chronic wounds. By illuminating the tissue surface with selected narrow wavelength bands (e.g., blue, green and red components) of light or detecting the reflectance of white light within several narrow bandwidths of the visible spectrum (e.g., selected wavelengths of peak absorption from the blood absorption spectrum of white light), the device may also be used to image the presence of blood and microvascular networks within and around the wound, including the surrounding normal tissue, thus also revealing areas of erythema and inflammation.
Reference is now made to
The regulation of angiogenesis over time during wound repair in vivo has been largely unexplored, due to difficulties in observing events within blood vessels. Although initial tests using an imaging device in accordance with the present disclosure were exploratory, simple modification of the device may allow longitudinal imaging of dynamic changes in blood supply and microvascular networks during the wound healing process in vivo.
In general, devices in accordance with the present disclosure may be used to image and/or monitor targets such as a skin target, an oral target, an ear-nose-throat target, an ocular target, a genital target, an anal target, and any other suitable targets on a subject.
Use in Clinical Care
Although current wound management practice aims to decrease the morbidity and mortality of wounds in patients, a limitation is the availability of health care resources. The potential of incorporating the technology of telemedicine into wound care needs is currently being explored. Wound care is a representation of the care of chronic and debilitating conditions that require long-term specialized care. The major effect of improved living conditions and advances in health care globally has led to people living longer. Therefore, the percentage of worlds' elderly and those with chronic medical conditions that would require medical attention is rising. With the escalating costs of health care, and the push of the industry towards outpatient care, this is a part of the health care crisis that is demanding immediate attention.
Devices in accordance with the present disclosure may provide biologically-relevant information about wounds and may exploit the emerging telemedicine (e.g., E-health) infrastructure to provide a solution for mobile wound care technology and may greatly impact wound health care treatment. Wound care accounts for a large percentage of home visits conducted by nurses and health care workers. Despite best practices some wounds do not heal as expected and require the services of a clinical specialist. The exemplary devices described herein may enable access to specialized clinical resources to help treat wounds from the convenience of the patient's home or chronic care facility, which decreases travel time for clients, increases availability to clinical wound specialists, and may reduce costs to the health care system.
Different uses of the imaging devices have been discussed for wound assessment, monitoring and care management. The devices may be used to detect and monitor changes in connective tissues (e.g., collagen, elastin) and blood/vascular supply during the wound healing process, monitor tissue necrosis and exudate in wounds based on fluorescence, detect and diagnose wound infections including potentially indicating critical ‘clinically significant’ categories of the presence of bacteria or micro-organisms (e.g., for detecting contamination, colonization, critical colonization and infection) at the surface and deep within wounds, provide topographic information of the wound, and identify wound margins and surrounding normal tissues. Tissue fluorescence and reflectance imaging data may be ‘mapped’ onto the white light images of the wound thereby permitting visualization within the wound and the surrounding normal tissues of essential wound biochemical and photobiological (e.g., fluorescence) information, which has not been possible to date. Real-time imaging of wounds may be performed over time to monitor changes in wound healing, and to potentially monitor the effectiveness of treatments by providing useful information about underlying biological changes that are occurring at the tissue/cellular level (e.g., matrix remodeling, inflammation, infection and necrosis). This may provide quantitative and objective wound information for detection, diagnosis and treatment monitoring in patients. In particular, such devices may be used to monitor and/or track the effectiveness of therapy at a biological level (e.g., on a bacterial level), which may provide more information than monitoring only the macroscopic/morphological appearance using white light.
The devices may provide real-time non-invasive image-guided biopsy targeting, clinical procedural guidance, tissue characterization, and may enable image-guided treatment using conventional and emerging modalities (e.g., PDT). In addition, use of the imaging devices may be used to correlate critical biological and molecular wound information obtained by fluorescence (e.g., endogenous tissue autofluorescence and/or administration of exogenous molecular-biomarker targeted fluorescence contrast agents) with existing and emerging clinical wound care assessment and treatment guides, such as the NERDS and STONES guidelines proposed by Sibbald et al. (Sibbald et al. Increased Bacterial Burden and Infection: The Story of NERDS and STONES. ADV SKIN WOUND CARE 2006; 19:447-61). The fluorescence imaging data obtained with the devices may be used to characterize, spatially and spectrally, bacterial balance and burden at the superficial and deep levels of wounds. The devices may provide real-time non-invasive image-guided biopsy targeting, clinical procedural guidance, tissue characterization, and may enable image-guided treatment using conventional and emerging treatment modalities (e.g., photodynamic therapy, PDT). The devices may be used within the clinical setting and integrated into conventional clinical wound care regimens, and may have a distinct role in areas of infectious diseases. It should be noted as well that such devices may also be used for real-time analysis, monitoring and care for chronic and acute wounds in animals and pets, via conventional veterinary care.
Devices in accordance with the present disclosure may allow real-time wound healing assessment for a large patient cohort base. In particular, elderly people, diabetics, immuno-suppressed and immobilized individuals have an increased incidence of chronic wounds and other dermal afflictions that result from poor circulation and immobility, e.g. pressure ulcers such as bed sores, venous stasis ulcers, and diabetic ulcers. These chronic conditions greatly increase the cost of care and reduce the patient's quality of life. As these groups are growing in number, the need for advanced wound care products will increase. Such devices may impact patient care by allowing a cost-effective means of monitoring chronic and acute wounds in a number of settings, including hospitals, ambulatory clinics, chronic care facilities, in-home-visit health care, emergency rooms and other critical areas in health care facilities. Further, such ‘handheld’ and portable imaging devices may be easily carried and used by nursing and ambulance staff. Early identification of scarring, which is related to connective tissue production and re-modeling of the wound, and bacterial infections may be detected and treated appropriately, something that is currently difficult. In addition, recent developments in advanced wound-care products including multiple dressing types (e.g., film, hydrocolloid, foam, anti-microbial, alginate, non-adherent, impregnated), hydrogels, wound cleansers and debriding agents, tissue engineered products (e.g., skin replacements, substitutes, and tissue-engineered products such as synthetic polymer-based biological tissue and growth factors), wound cleansers, pharmacological products, and physical therapies may also benefit from the device developed here as it may allow image-based longitudinal monitoring of the effectiveness of such treatments. Physical therapies may include hydrotherapy, electrical stimulation, electromagnetic stimulation devices, ultraviolet therapy, hyperbaric oxygen therapy, ultrasound devices, laser/light emitting diode (LED) devices, and wound imaging/documentation. Additional therapies may include, for example, antibiotics, wound debridement, application of wound dressings, and wound cleaning.
Wound tissue analysis is typically required for the assessment of the healing of skin wounds. Percentage of the granulation tissue, fibrin and necrosis in the wound, and their change during treatment may provide useful information that may guide wound treatment. Image analysis may include advanced statistical pattern recognition and classification algorithms to identify individual pixels within the fluorescence wound images collected with the device based on the optical information of the wound and surrounding normal tissue. Thus, image analysis may allow wound images to be mapped into various components of the wound, including total wound area, epithelialization, granulation, slough, necrotic, hypergranulation, infected, undermining, and surrounding tissue margins. This has an added advantage of providing relatively rapid determination of wound healing rates, as well as informing guide patient management decisions.
This device may be easily integrated into existing health-care computer infrastructures (e.g., desktop and pocket PCs used by a growing number of physicians or other health care professionals) for longitudinal image cataloguing for patient wound management within the conventional clinical environment. The wireless receiving and transmission of data capabilities of the device may allow monitoring of wound care and healing remotely through existing and future wireless telemedicine infrastructure. The device may be used to transfer essential medical data (e.g., wound health status) via the internet or over wireless services, such as cellular telephone, PDA or Smartphone services, to remote sites which may permit remote medical interventions, with a further utility in military medical applications for battlefield wound management. The device may allow real-time surface imaging of wound sites and may be easily carried by point-of-care personnel in clinical settings. Using cost-effective highly sensitive commercially available digital imaging devices, such as digital cameras, cellular phones, PDAs, laptop computers, tablet PCs, webcams, and Smart phones, etc. as the image capture or recording component, the device may offer image-based documentation of wound healing and tracking of treatment effectiveness. Also, this technology may be adapted to also function in ‘wireless’ mode to permit remote medical interventions by potentially adapting it for use with high-resolution digital cameras embedded in commercially-available cellular telephones.
By using web-based telemedicine and remote medical monitoring infrastructure, the imaging device may be integrated into a ‘store-and-forward’ concept of wound assessment systems. In addition to providing digital images, such a system may present a comprehensive set of clinical data that meet the recommendations of clinical practice guidelines. The presently-disclosed devices may integrate into a computer-based wound assessment system (e.g., with image analysis software) to be used by a health care facility to enhance existing clinical databases and support the implementation of evidence-based practice guidelines. Such an integrated telemedicine infrastructure may be used for monitoring patients at home or in long-term-care facilities, who may benefit from routine monitoring by qualified clinicians but currently do not have access to this care. These devices may be further developed into a portable handheld point-of-care diagnostic system, which may represent a major advance in detecting, monitoring, treating, and preventing infectious disease spread in the developed and developing worlds. This knowledge may significantly improve the diagnostic tools available to practitioners who treat chronic wounds in settings where quantitative cultures are inaccessible.
Devices in accordance with the present disclosure may allow digital imaging with optical and digital zooming capabilities (e.g., those embedded in commonly available digital imaging devices). Still or video image quality may be in ‘high-definition’ format to achieve high spatial resolution imaging of the tissue surface. Images may be recorded as still/freeze frame and/or in video/movie format and printed using standard imaging printing protocols which do (e.g., connected via USB) or do not (e.g., PictBridge) require a personal computer. The images/video data may be transferred to a personal computer for data archival storage and/or image viewing and/or analysis/manipulation. Such devices may also transfer data to a printer or personal computer using wired or wireless capabilities (e.g., Bluetooth). Visualization may be performed on the handheld device screen and/or in addition to simultaneous viewing on a video screen/monitor (e.g., head-mounted displays and glasses) using standard output video cables. These devices may display, in combination or separately, optical wavelength and fluorescence/reflectance intensity information with spatial dimensions of the imaged scene to allow quantitative measurements of distances (e.g., monitoring changes tissue morphology/topography) over time. The devices may also allow digital image/video storage/cataloguing of images and related patient medical data, for example using dedicated software with imaging analysis capabilities and/or diagnostic algorithms.
Image Analysis
Image analysis may be used together with the exemplary devices of the present disclosure to quantitatively measure fluorescence intensities and relative changes in multiple fluorescence spectra (e.g., multiplexed imaging) of the exogenous optical molecular targeting probes in the wound and surrounding normal tissues. The biodistributions of the fluorescent probes may be determined based on the fluorescence images collected and these may be monitored over time between individual clinical wound imaging sessions for change. By determining the presence and relative changes in abundance quantitatively, using the devices, of each and all of the spectrally-unique fluorescent probes, the clinical operator may determine in real-time or near real-time the health and/or healing status and response to treatment over time of a given wound, for example by using a look-up table in which specific tissue, cellular and molecular signals are displayed in correlation to wound health, healing and response status, an example of which is shown in
Image analysis techniques may be used to calibrate the initial or first images of the wound using a portable fluorescent standard placed within the field of view during imaging with a device. The image analysis may also permit false or pseudo color display on a monitor for differentiating different biological (e.g., tissue, cellular, and molecular) components of the wound and surrounding normal tissues including those biomarkers identified by autofluorescence and those identified by the use of exogenous targeted or untargeted fluorescence/absorption contrast agents.
Examples of such biomarkers are listed in
Pre-assigned color maps may be used to display simultaneously the biological components of the wound and surrounding normal tissues including connective tissues, blood, microvascularity, bacteria, microorganisms, etc. as well as fluorescently labeled drugs/pharmacological agents. This may permit visualization in real-time or near real-time (e.g., less than 1 minute) of the health, healing and infectious status of the wound area.
The image analysis algorithms may provide one or more of the following features:
Patient Digital Image Management
Although image analysis algorithm, techniques, or software have been described, this description also extends to a computing device, a system, and a method for carrying out this image analysis.
Image-Guidance
Devices in accordance with the present disclosure may also be useful for providing fluorescent image-guidance, for example in surgical procedures, even without the use of dyes or markers. Certain tissues and/or organs may have different fluorescent spectra (e.g., endogenous fluorescence) when viewed using the imaging device, or example under certain excitation light conditions.
Bioengineered Skin
Several bioengineered skin products or skin equivalents have become available commercially for the treatment of acute and chronic wounds, as well as burn wounds. These have been developed and tested in human wounds. Skin equivalents may contain living cells, such as fibroblasts or keratinocytes, or both, while others are made of acellular materials or extracts of living cells. The clinical effect of these constructs is 15-20% better than conventional ‘control’ therapy, but there is debate over what constitutes an appropriate control. Bioengineered skin may work by delivering living cells which are known as a ‘smart material’ because they are capable of adapting to their environment. There is evidence that some of these living constructs are able to release growth factors and cytokines. Exogenous fluorescent molecular agents may be used in conjunction with such skin substitutes to determine completeness of engraftment as well as biological response of the wound to the therapy. The healing of full-thickness skin defects may require extensive synthesis and remodeling of dermal and epidermal components. Fibroblasts play an important role in this process and are being incorporated in the latest generation of artificial dermal substitutes.
The exemplary imaging devices described herein may be used to determine the fate of fibroblasts seeded in skin substitute and the influence of the seeded fibroblasts on cell migration and dermal substitute degradation after transplantation to a wound site. Wounds may be treated with either dermal substitutes seeded with autologous fibroblasts or acellular substitutes. Seeded fibroblasts, labeled with a fluorescent cell marker, may then be detected in the wounds with a fluorescence imaging device and then quantitatively assessed using image analysis, for example as described above.
Polymer-Based Therapeutic Agents
There are a number of commercially available medical polymer products made for wound care. For example, Rimon Therapeutics produces Theramers™ (www.rimontherapeutics.com) which are medical polymers that have biological activity in and of themselves, without the use of drugs. Rimon Therapeutics produces the following wound care products, which can be made to be uniquely fluorescent, when excited by 405 nm excitation light: Angiogenic Theramer™, which induces new blood vessel development (i.e., angiogenesis) in wounds or other ischemic tissue; MI Theramer™, which inhibits the activity of matrix metalloproteases (MMPs), a ubiquitous group of enzymes that are implicated in many conditions in which tissue is weakened or destroyed; AM Theramer™, a thermoplastic that kills gram positive and gram negative bacteria without harming mammalian cells; and ThermaGel™, a polymer that changes from a liquid to a strong gel reversibly around body temperature. These can each be made to be fluorescent by addition of fluorescent dyes or fluorescent nanoparticles selected to be excited, for example, at 405 nm light with longer wavelength fluorescence emission.
By using the exemplary imaging devices of the present disclosure, the application of such fluorescent polymer agents may be guided by fluorescent imaging in real-time. This may permit the Theramer agent to be accurately delivered/applied (e.g., topically) to the wound site. Following application of the agent to the wound, a fluorescent imaging device may then be used to quantitatively determine the therapeutic effects of the Theramers on the wound as well as track the biodistribution of these in the wound over time, in vivo and non-invasively. It may also be possible to add a molecular beacon, possibly having another fluorescent emission wavelength, to the MI Theramer™ that can fluoresce in the presence of wound enzymes (e.g., MMPs), and this may indicate in real-time the response of the wound to the MI Theramer™. It may be possible to use one fluorescence emission for image-guided Theramer application to the wound site and another different fluorescence emission for therapeutic response monitoring, and other fluorescence emissions for other measurements. The relative effectiveness of MMP inhibition and antimicrobial treatments may be determined simultaneously over time. Using image analysis, real-time comparison of changes in fluorescence of these signals in the wound may be possible. This adds a quantitative aspect to the device and adds to its clinical usefulness.
It should be noted that other custom bio-safe fluorescence agents may be added to the following materials which are currently used for wound care. The fluorescent material may then be imaged and monitored using the disclosed devices.
Other wound care products that may be imaged using the disclosed devices include Theramers, silver-containing gels (e.g., hydrogels), artificial skin, ADD stem cells, anti-matrix metalloproteinases, and hyaluronic acid. Fluorescent agents may be added to other products to allow for imaging using the devices. In some cases, the products may already be luminescent and may not require the addition of fluorescent agents.
The exemplary disclosed devices may be used also to monitor the effects of such treatments over time.
Cosmetic Applications
Device in accordance with the present disclosure may be used to image a patient's skin surface. For example, the device may be used to obtain images of the patient's skin by detecting autofluorescence produced by violet/blue light excitation of the skin surface. Red fluorescence from P. acnes may be easily detected in regions of the patient's face. P. acnes is the causative agent of acne vulgaris (i.e., pimples) and is a common resident of the pilosebaceous glands of the human skin. Furthermore, P. acnes is occult under white light visualization. The auto fluorescent images of the patient's skin may be obtained without the need of exogenous agents/drugs and demonstrate the capability of the device to detect bacteria in single skin pores.
Such a capability to image and document the presence and biodistribution of bacteria on the skin surface makes the device potentially useful in the dermatology and cosmetology fields. For example, fluorescence imaging may be performed prior to, during, and after application of dermatological treatment and/or pharmaceutical/cosmetic formulations (e.g., topical creams, drugs and other antibiotics, skin disinfecting agents, acne treatments, etc.) to the normal and abnormal skin conditions, including but not limited to scarring, hyper-pigmentation, acne, psoriasis, eczema, rashes, etc. Fluorescence/reflectance image-guided tattoo removal (e.g., using surgery or available laser treatments) may also be an option with the device.
The device was also used to image minor cuts, scrapes, and abrasions on patients' skin and under violet/blue light. Tissue autofluorescence from connective tissue components (e.g., collagen and elastin) from the wound site and surrounding normal skin aided in detecting white light-occult changes in connective tissues during minor cutaneous wound healing (as seen in
With reference now to
The devices may also be used to image, assess and longitudinally monitor the healing process in burns or determine the response of skin grafts or temporary skin substitutes in treatment of burn patients. The devices may also serve to detect and monitor late radiation-induced skin damage during treatment of patients with ionizing radiation.
Darkening Environment
In accordance with various embodiments of the present disclosure, it may be beneficial to use the disclosed imaging devices in a reduced lighting environment, such as a dimly lit environment or completely dark environment, to obtain FL images. In cases where ambient lighting cannot be dimmed, reduced sufficiently, or completely turned off, the optimal lighting conditions can be managed with optical accessories, such as a tent or drape. For example, when using the device of the present disclosure in a lit environment (such as in an operating room), a tent or drape may be used to create a darkened (dimly lit or complete dark) environment around the imaging target, for example around a limb of a patient. In some embodiments, the device includes a mechanism to allow for attachment of the tent or drape to the imaging device. The tent or drape may be disposable and may be packaged with the device as part of a system.
In some embodiments, the imaging device in accordance with the present disclosure may be used in a room (such as an operating room) and one or more lights in the room may be turned off to produce the dimly lit or completely dark environment required for fluorescence imaging. In this embodiment, the imaging device may also be used with the tent or drape.
In some exemplary embodiments, the device may also be configured to prompt a user to confirm that the lighting conditions in the environment are sufficient (i.e., dimmed/darkened) when enabling the FL functionality of the imaging device. In other exemplary embodiments, the device may also display an indicator, such as, for example a moon icon on the image, to denote that the image was taken in an appropriate environment (i.e., in a dimmed and/or darkened environment).
Kits for Device
Imaging devices in accordance with the present disclosure also may be provided in a kit, for example including the device and a fluorescing contrast agent. The contrast agent may be any one or more of those described above. For example, the contrast agent may be for labeling a biomarker in a wound, where the kit is for wound monitoring applications. Alternatively, the imaging device and a drape may be packaged or otherwise provided together.
The imaging device may be used in white light and fluorescence modes to improve the administration of these treatments as well as monitor their effectiveness over time non-invasively and quantitatively. The device may be used in combination with other imaging modalities, for example thermal imaging methods, among others.
While the present disclosure has been disclosed in terms of exemplary embodiments in order to facilitate better understanding of the disclosure, it should be appreciated that the disclosure can be embodied in various ways without departing from the principle of the disclosure. Therefore, the disclosure should be understood to include all possible embodiments which can be embodied without departing from the principle of the disclosure set out in the appended claims. Furthermore, although the present disclosure has been discussed with relation to wound imaging, monitoring, and analysis those of ordinary skill in the art would understand that the present teachings as disclosed would work equally well in various other applications such as, for example, clinically- and research-based imaging of small and large (e.g., veterinary) animals; detection and monitoring of contamination (e.g., bacterial contamination) in food/animal product preparation in the meat, poultry, dairy, fish, agricultural industries; detection of ‘surface contamination’ (e.g., bacterial or biological contamination) in public (e.g., health care) and private settings; multi-spectral imaging and detection of cancers in human and/or veterinary patients; as a research tool for multi-spectral imaging and monitoring of cancers in experimental animal models of human diseases (e.g., wound and cancers); forensic detection, for example of latent finger prints and biological fluids on non-biological surfaces; imaging and monitoring of dental plaques, carries and cancers in the oral cavity; periodontal disease; cancers in the tongue; imaging of an ear-nose-throat target, imaging of an ocular target; imaging of a genital target; imaging of an anal target; imaging of other suitable targets on a subject; burn wounds; imaging and monitoring device in clinical microbiology laboratories; and testing anti-bacterial (e.g., antibiotic), disinfectant agents. The use of a fluorescent imaging device in such environments is disclosed in U.S. Pat. No. 9,042,967 B2 to DaCosta et al., entitled “Device and Method for Wound Imaging and Monitoring,” and issued on May 26, 2015, which is incorporated by reference herein. Additionally or alternatively, the device may be used for detecting and imaging of the presence of bacteria or microbes and other pathogens on a variety of surfaces, materials, instruments (e.g., surgical instruments) in hospitals, chronic care facilities, old age homes, and other health care settings where contamination may be the leading source of infection. The device may be used in conjunction with standard detection, identification and enumeration of indicator organisms and pathogens strategies.
For the purposes of this specification and appended claims, unless otherwise indicated, all numbers expressing quantities, percentages or proportions, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the written description and claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.
It is noted that, as used in this specification and the appended claims, the singular forms “a,” “an,” and “the,” include plural referents unless expressly and unequivocally limited to one referent. Thus, for example, reference to “a sensor” includes two or more different sensors. As used herein, the term “include” and its grammatical variants are intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that can be substituted or added to the listed items.
It will be apparent to those skilled in the art that various modifications and variations can be made to the system and method of the present disclosure without departing from the scope its teachings. Other embodiments of the disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the teachings disclosed herein. It is intended that the specification and embodiment described herein be considered as exemplary only.
This application is a continuation of U.S. application Ser. No. 16/593,174, filed Oct. 4, 2019, which is a continuation of U.S. application Ser. No. 15/328,214, filed Jan. 23, 2017, which is a U.S. national stage application under 35 U.S.C. § 371(c) of International Application No. PCT/CA2015/000444, filed on Jul. 24, 2015, which claims priority to U.S. Provisional Application No. 62/028,386, filed Jul. 24, 2014, the entire contents of each of which are incorporated by reference herein.
| Number | Name | Date | Kind |
|---|---|---|---|
| 4556057 | Hiruma et al. | Dec 1985 | A |
| 4740459 | Chen et al. | Apr 1988 | A |
| 5306144 | Hibst et al. | Apr 1994 | A |
| 5421337 | Richards-Kortum et al. | Jun 1995 | A |
| 5456260 | Kollias et al. | Oct 1995 | A |
| 5474910 | Alfano | Dec 1995 | A |
| 5482041 | Wilk et al. | Jan 1996 | A |
| 5507287 | Palcic et al. | Apr 1996 | A |
| 5522868 | Buckley et al. | Jun 1996 | A |
| 5533508 | Doiron | Jul 1996 | A |
| 5552134 | Morgan et al. | Sep 1996 | A |
| 5562100 | Kittrell et al. | Oct 1996 | A |
| 5566673 | Shiono et al. | Oct 1996 | A |
| 5569911 | Tomlinson, Jr. et al. | Oct 1996 | A |
| 5572996 | Poiron et al. | Nov 1996 | A |
| 5579773 | Vo-Dinh et al. | Dec 1996 | A |
| 5588428 | Smith et al. | Dec 1996 | A |
| 5590660 | MacAulay et al. | Jan 1997 | A |
| 5605152 | Slate et al. | Feb 1997 | A |
| 5612540 | Richards-Kortum et al. | Mar 1997 | A |
| 5623932 | Ramanujam et al. | Apr 1997 | A |
| 5626134 | Zuckerman | May 1997 | A |
| 5628310 | Rao et al. | May 1997 | A |
| 5647368 | Zeng et al. | Jul 1997 | A |
| 5687730 | Doiron et al. | Nov 1997 | A |
| 5690417 | Polidor et al. | Nov 1997 | A |
| 5697373 | Richards-Kortum et al. | Dec 1997 | A |
| 5701902 | Vari et al. | Dec 1997 | A |
| 5760407 | Margosiak et al. | Jun 1998 | A |
| 5769792 | Palcic et al. | Jun 1998 | A |
| 5770454 | Essenpreis et al. | Jun 1998 | A |
| 5779631 | Chance | Jul 1998 | A |
| 5813987 | Modell et al. | Sep 1998 | A |
| 5820558 | Chance | Oct 1998 | A |
| 5849595 | Alfano et al. | Dec 1998 | A |
| 5851181 | Talmor | Dec 1998 | A |
| 5865754 | Sevick-Muraca et al. | Feb 1999 | A |
| 5879294 | Anderson et al. | Mar 1999 | A |
| 5935075 | Casscells et al. | Aug 1999 | A |
| 5952664 | Wake et al. | Sep 1999 | A |
| 5981958 | Li et al. | Nov 1999 | A |
| 5986271 | Lazarev et al. | Nov 1999 | A |
| 6002137 | Hayashi | Dec 1999 | A |
| 6008889 | Zeng et al. | Dec 1999 | A |
| 6014204 | Prahl et al. | Jan 2000 | A |
| 6021344 | Lui et al. | Feb 2000 | A |
| 6026319 | Hayashi | Feb 2000 | A |
| 6036941 | Bottiroli et al. | Mar 2000 | A |
| 6055451 | Bambot et al. | Apr 2000 | A |
| 6058324 | Chance | May 2000 | A |
| 6061591 | Freitag et al. | May 2000 | A |
| 6064897 | Lindberg et al. | May 2000 | A |
| 6064899 | Fein et al. | May 2000 | A |
| 6069689 | Zeng et al. | May 2000 | A |
| 6078833 | Hueber | Jun 2000 | A |
| 6081612 | Gutkowicz-Krusin et al. | Jun 2000 | A |
| 6081739 | Lemchen | Jun 2000 | A |
| 6088087 | Graves et al. | Jul 2000 | A |
| 6088606 | Ignotz et al. | Jul 2000 | A |
| 6091984 | Perelman et al. | Jul 2000 | A |
| 6091985 | Alfano et al. | Jul 2000 | A |
| 6095982 | Richards-Kortum et al. | Aug 2000 | A |
| 6104939 | Groner et al. | Aug 2000 | A |
| 6104945 | Modell et al. | Aug 2000 | A |
| 6122042 | Wunderman et al. | Sep 2000 | A |
| 6122536 | Sun et al. | Sep 2000 | A |
| 6124597 | Shehada et al. | Sep 2000 | A |
| 6128516 | Macfarlane et al. | Oct 2000 | A |
| 6128525 | Zeng et al. | Oct 2000 | A |
| 6129664 | Macfarlane et al. | Oct 2000 | A |
| 6135965 | Tumer et al. | Oct 2000 | A |
| 6142629 | Adel et al. | Nov 2000 | A |
| 6148227 | Wagnieres et al. | Nov 2000 | A |
| 6161031 | Hochman et al. | Dec 2000 | A |
| 6167297 | Benaron | Dec 2000 | A |
| 6192261 | Gratton et al. | Feb 2001 | B1 |
| 6205354 | Gellermann et al. | Mar 2001 | B1 |
| 6208749 | Gutkowicz-Krusin et al. | Mar 2001 | B1 |
| 6212425 | Irion et al. | Apr 2001 | B1 |
| 6219566 | Weersink et al. | Apr 2001 | B1 |
| 6223071 | Lundahl et al. | Apr 2001 | B1 |
| 6236871 | Tsuchiya | May 2001 | B1 |
| 6236881 | Zahler et al. | May 2001 | B1 |
| 6238348 | Crowley et al. | May 2001 | B1 |
| 6241662 | Richards-Kortum et al. | Jun 2001 | B1 |
| 6256530 | Wolfe | Jul 2001 | B1 |
| 6258576 | Richards-Kortum et al. | Jul 2001 | B1 |
| 6272367 | Chance | Aug 2001 | B1 |
| 6272376 | Marcu et al. | Aug 2001 | B1 |
| 6280386 | Alfano et al. | Aug 2001 | B1 |
| 6289236 | Koenig et al. | Sep 2001 | B1 |
| 6304766 | Colvin, Jr. | Oct 2001 | B1 |
| 6317624 | Kollias et al. | Nov 2001 | B1 |
| 6343227 | Crowley | Jan 2002 | B1 |
| 6364829 | Fulghum | Apr 2002 | B1 |
| 6373568 | Miller et al. | Apr 2002 | B1 |
| 6385484 | Nordstrom et al. | May 2002 | B2 |
| 6393315 | Aprahamian et al. | May 2002 | B1 |
| 6422994 | Kaneko et al. | Jul 2002 | B1 |
| 6427082 | Nordstrom et al. | Jul 2002 | B1 |
| 6436127 | Anderson et al. | Aug 2002 | B1 |
| 6442416 | Schultz | Aug 2002 | B1 |
| 6465968 | Sendai | Oct 2002 | B1 |
| 6473637 | Hayashi | Oct 2002 | B1 |
| 6487428 | Culver et al. | Nov 2002 | B1 |
| 6496719 | Hayashi | Dec 2002 | B2 |
| 6510338 | Irion et al. | Jan 2003 | B1 |
| 6522911 | Toida et al. | Feb 2003 | B1 |
| 6526309 | Chance | Feb 2003 | B1 |
| 6542772 | Chance | Apr 2003 | B1 |
| 6549801 | Chen et al. | Apr 2003 | B1 |
| 6563105 | Seibel et al. | May 2003 | B2 |
| 6571119 | Hayashi | May 2003 | B2 |
| 6573513 | Hayashi | Jun 2003 | B2 |
| 6574502 | Hayashi | Jun 2003 | B2 |
| 6577394 | Zavislan | Jun 2003 | B1 |
| 6577884 | Boas | Jun 2003 | B1 |
| 6580941 | Webb | Jun 2003 | B2 |
| 6582363 | Adachi et al. | Jun 2003 | B2 |
| 6584342 | Trushin et al. | Jun 2003 | B1 |
| 6590651 | Bambot et al. | Jul 2003 | B1 |
| 6597932 | Tian et al. | Jul 2003 | B2 |
| 6600947 | Averback et al. | Jul 2003 | B2 |
| 6603126 | Yamada et al. | Aug 2003 | B2 |
| 6603552 | Cline et al. | Aug 2003 | B1 |
| 6615063 | Ntziachristos et al. | Sep 2003 | B1 |
| 6622032 | Robinson et al. | Sep 2003 | B1 |
| 6631286 | Pfeiffer et al. | Oct 2003 | B2 |
| 6631289 | Alfano et al. | Oct 2003 | B2 |
| 6636759 | Robinson | Oct 2003 | B2 |
| 6639668 | Trepagnier | Oct 2003 | B1 |
| 6640124 | Elsner et al. | Oct 2003 | B2 |
| 6640130 | Freeman et al. | Oct 2003 | B1 |
| 6640131 | Irion et al. | Oct 2003 | B1 |
| 6652836 | Luiken | Nov 2003 | B2 |
| 6665556 | Alfano et al. | Dec 2003 | B1 |
| 6667803 | Flessland et al. | Dec 2003 | B1 |
| 6668186 | Zavislan et al. | Dec 2003 | B1 |
| 6678398 | Wolters et al. | Jan 2004 | B2 |
| 6690466 | Miller et al. | Feb 2004 | B2 |
| 6694158 | Polak | Feb 2004 | B2 |
| 6697652 | Georgakoudi et al. | Feb 2004 | B2 |
| 6697657 | Shehada et al. | Feb 2004 | B1 |
| 6697666 | Richards-Kortum et al. | Feb 2004 | B1 |
| 6701168 | Wilson et al. | Mar 2004 | B1 |
| 6709446 | Lundahl et al. | Mar 2004 | B2 |
| 6711429 | Gilboa et al. | Mar 2004 | B1 |
| 6721582 | Trepagnier et al. | Apr 2004 | B2 |
| 6730113 | Eckhardt et al. | May 2004 | B2 |
| 6738659 | Hsu | May 2004 | B2 |
| 6748259 | Benaron et al. | Jun 2004 | B1 |
| 6750964 | Levenson et al. | Jun 2004 | B2 |
| 6751490 | Esenaliev et al. | Jun 2004 | B2 |
| 6757554 | Rubinstein et al. | Jun 2004 | B2 |
| 6763262 | Hohla et al. | Jul 2004 | B2 |
| 6766184 | Utzinger et al. | Jul 2004 | B2 |
| 6768918 | Zelenchuk | Jul 2004 | B2 |
| 6778846 | Martinez et al. | Aug 2004 | B1 |
| 6800057 | Tsujita et al. | Oct 2004 | B2 |
| 6806089 | Lakowicz et al. | Oct 2004 | B1 |
| 6818903 | Schomacker et al. | Nov 2004 | B2 |
| 6821289 | Bode et al. | Nov 2004 | B2 |
| 6826424 | Zeng et al. | Nov 2004 | B1 |
| 6865407 | Kimball et al. | Mar 2005 | B2 |
| 6868285 | Muller-Dethlefs | Mar 2005 | B2 |
| 6870620 | Faupel et al. | Mar 2005 | B2 |
| 6873716 | Bowker et al. | Mar 2005 | B1 |
| 6912412 | Georgakoudi et al. | Jun 2005 | B2 |
| 6914250 | Seville | Jul 2005 | B2 |
| 6922523 | Merola et al. | Jul 2005 | B2 |
| 6922576 | Raskas | Jul 2005 | B2 |
| 6930773 | Cronin et al. | Aug 2005 | B2 |
| 6933154 | Schomacker et al. | Aug 2005 | B2 |
| 6936004 | Utsui | Aug 2005 | B2 |
| 6970729 | Hartmann | Nov 2005 | B2 |
| 6975898 | Seibel | Dec 2005 | B2 |
| 6975899 | Faupel et al. | Dec 2005 | B2 |
| 6984228 | Anderson et al. | Jan 2006 | B2 |
| 6992762 | Long et al. | Jan 2006 | B2 |
| 7006223 | Mullani | Feb 2006 | B2 |
| 7016714 | Colvin, Jr. | Mar 2006 | B2 |
| 7016717 | Demos et al. | Mar 2006 | B2 |
| 7027153 | Mullani | Apr 2006 | B2 |
| 7062311 | Sendai et al. | Jun 2006 | B1 |
| 7102142 | Sendai | Sep 2006 | B2 |
| 7103401 | Schomacker et al. | Sep 2006 | B2 |
| 7107116 | Geng | Sep 2006 | B2 |
| 7113814 | Ward et al. | Sep 2006 | B2 |
| 7127282 | Nordstrom et al. | Oct 2006 | B2 |
| 7130672 | Pewzner et al. | Oct 2006 | B2 |
| 7139598 | Hull et al. | Nov 2006 | B2 |
| 7139603 | Chance | Nov 2006 | B2 |
| 7149567 | Demos et al. | Dec 2006 | B2 |
| 7151270 | Birk et al. | Dec 2006 | B2 |
| 7167243 | Mullani | Jan 2007 | B2 |
| 7167244 | Mullani | Jan 2007 | B2 |
| 7197355 | Nelson | Mar 2007 | B2 |
| 7202947 | Liu et al. | Apr 2007 | B2 |
| 7209773 | Juliano | Apr 2007 | B2 |
| 7212848 | Wake et al. | May 2007 | B1 |
| 7218822 | Treado et al. | May 2007 | B2 |
| 7224468 | Fouquet | May 2007 | B2 |
| RE39672 | Shehada et al. | Jun 2007 | E |
| 7236815 | Richards-Kortum et al. | Jun 2007 | B2 |
| 7242997 | Geng | Jul 2007 | B2 |
| 7248182 | Dudda et al. | Jul 2007 | B2 |
| 7257437 | Demos et al. | Aug 2007 | B2 |
| 7277210 | Lipson | Oct 2007 | B2 |
| 7282723 | Schomacker et al. | Oct 2007 | B2 |
| 7283858 | Sendai | Oct 2007 | B2 |
| 7286224 | Curry et al. | Oct 2007 | B2 |
| 7289836 | Colvin, Jr. | Oct 2007 | B2 |
| 7310547 | Zelenchuk | Dec 2007 | B2 |
| 7317938 | Lorenz et al. | Jan 2008 | B2 |
| 7321791 | Levenson et al. | Jan 2008 | B2 |
| 7324608 | Chiodini et al. | Jan 2008 | B1 |
| 7353055 | Hogan | Apr 2008 | B2 |
| 7359748 | Drugge | Apr 2008 | B1 |
| 7365844 | Richards-Kortum et al. | Apr 2008 | B2 |
| 7366365 | Carver | Apr 2008 | B2 |
| 7368694 | Goulas et al. | May 2008 | B2 |
| 7376346 | Merola et al. | May 2008 | B2 |
| 7376451 | Mahony et al. | May 2008 | B2 |
| 7378056 | Black | May 2008 | B2 |
| 7389132 | Wang et al. | Jun 2008 | B2 |
| 7403812 | Rice et al. | Jul 2008 | B2 |
| 7404929 | Fulghum, Jr. | Jul 2008 | B2 |
| 7454046 | Chhibber et al. | Nov 2008 | B2 |
| 7477393 | Sendai | Jan 2009 | B2 |
| 7477767 | Chhibber et al. | Jan 2009 | B2 |
| 7477931 | Hoyt | Jan 2009 | B2 |
| 7479990 | Imaizumi et al. | Jan 2009 | B2 |
| 7491956 | Knoche et al. | Feb 2009 | B2 |
| 7495233 | Pfister et al. | Feb 2009 | B2 |
| 7496392 | Alarcon et al. | Feb 2009 | B2 |
| 7499161 | Richards-Kortum et al. | Mar 2009 | B2 |
| 7505809 | Strommer et al. | Mar 2009 | B2 |
| 7508524 | Mahadevan-Jansen et al. | Mar 2009 | B2 |
| 7510699 | Black et al. | Mar 2009 | B2 |
| 7515952 | Balas et al. | Apr 2009 | B2 |
| 7519411 | Long | Apr 2009 | B2 |
| 7522797 | Treado et al. | Apr 2009 | B2 |
| 7526329 | Hogan et al. | Apr 2009 | B2 |
| 7536213 | Lipson et al. | May 2009 | B2 |
| 7539530 | Caplan et al. | May 2009 | B2 |
| 7555332 | Rice et al. | Jun 2009 | B2 |
| 7558619 | Ferguson et al. | Jul 2009 | B2 |
| 7564550 | Yaroslavsky et al. | Jul 2009 | B2 |
| 7570359 | Fox | Aug 2009 | B2 |
| 7570984 | Katsuda et al. | Aug 2009 | B2 |
| 7580185 | Haisch et al. | Aug 2009 | B2 |
| 7583993 | Sendai | Sep 2009 | B2 |
| 7587236 | Demos et al. | Sep 2009 | B2 |
| 7590437 | Rubinstein et al. | Sep 2009 | B2 |
| 7598088 | Balas | Oct 2009 | B2 |
| 7599065 | Sendai | Oct 2009 | B2 |
| 7599731 | Rice et al. | Oct 2009 | B2 |
| 7599732 | Sevick-Muraca et al. | Oct 2009 | B2 |
| 7603031 | Viaud et al. | Oct 2009 | B1 |
| 7609814 | Baumgart | Oct 2009 | B2 |
| 7610082 | Chance | Oct 2009 | B2 |
| 7613504 | Rowe | Nov 2009 | B2 |
| 7613505 | Mazuir et al. | Nov 2009 | B2 |
| 7633621 | Thornton | Dec 2009 | B2 |
| 7646002 | Sendai | Jan 2010 | B2 |
| 7652763 | Matousek et al. | Jan 2010 | B2 |
| 7653424 | March | Jan 2010 | B2 |
| 7672702 | Hwang et al. | Mar 2010 | B2 |
| 7697966 | Monfre et al. | Apr 2010 | B2 |
| 7697975 | Zeng | Apr 2010 | B2 |
| 7702381 | Gaeta et al. | Apr 2010 | B2 |
| 7722537 | Sterling et al. | May 2010 | B2 |
| 7729732 | Ohashi | Jun 2010 | B2 |
| 7729749 | Roessler et al. | Jun 2010 | B2 |
| 7734325 | Vizard et al. | Jun 2010 | B2 |
| 7738032 | Kollias et al. | Jun 2010 | B2 |
| 7749217 | Podhajsky | Jul 2010 | B2 |
| 7761126 | Gardner et al. | Jul 2010 | B2 |
| 7764303 | Pote et al. | Jul 2010 | B2 |
| 7785277 | Babaev et al. | Aug 2010 | B2 |
| 7787928 | Frisch et al. | Aug 2010 | B2 |
| 7794925 | Cullen | Sep 2010 | B2 |
| 7798955 | Ishihara et al. | Sep 2010 | B2 |
| 7804991 | Abovitz et al. | Sep 2010 | B2 |
| 7812945 | Fortier et al. | Oct 2010 | B2 |
| 7817267 | Carver | Oct 2010 | B2 |
| 7821640 | Koenig et al. | Oct 2010 | B2 |
| 7822450 | Colvin, Jr. et al. | Oct 2010 | B2 |
| 7826878 | Alfano et al. | Nov 2010 | B2 |
| 7843572 | Tearney et al. | Nov 2010 | B2 |
| 7860554 | Leonardi et al. | Dec 2010 | B2 |
| 7865231 | Tearney et al. | Jan 2011 | B2 |
| 7872759 | Tearney et al. | Jan 2011 | B2 |
| 7888659 | Scholz et al. | Feb 2011 | B2 |
| 7889348 | Tearney et al. | Feb 2011 | B2 |
| 7899518 | Trepagnier et al. | Mar 2011 | B2 |
| 7899624 | Cualing et al. | Mar 2011 | B2 |
| 7904140 | Pilon et al. | Mar 2011 | B2 |
| 7909253 | Sherman | Mar 2011 | B2 |
| 7912534 | Grinvald et al. | Mar 2011 | B2 |
| 7914442 | Gazdzinski | Mar 2011 | B1 |
| 7916283 | Fukutani et al. | Mar 2011 | B2 |
| 7925333 | Weir et al. | Apr 2011 | B2 |
| 7935055 | Burckhardt | May 2011 | B2 |
| 7945077 | Demos | May 2011 | B2 |
| 7960707 | Hall et al. | Jun 2011 | B2 |
| 7962200 | Ntziachristos et al. | Jun 2011 | B2 |
| 7966060 | Smit et al. | Jun 2011 | B2 |
| 7973925 | Lipson et al. | Jul 2011 | B2 |
| 7977650 | Laidevant et al. | Jul 2011 | B2 |
| 7979107 | Lin et al. | Jul 2011 | B2 |
| 7983736 | Villard et al. | Jul 2011 | B2 |
| 7983740 | Culver et al. | Jul 2011 | B2 |
| 7986342 | Yogesan et al. | Jul 2011 | B2 |
| 7986987 | Bazin et al. | Jul 2011 | B2 |
| 7996068 | Telischak et al. | Aug 2011 | B2 |
| 8000775 | Pogue et al. | Aug 2011 | B2 |
| 8005527 | Zelenchuk | Aug 2011 | B2 |
| 8026942 | Payonk et al. | Sep 2011 | B2 |
| 8031924 | Can et al. | Oct 2011 | B2 |
| 8039816 | Morishita et al. | Oct 2011 | B2 |
| 8041162 | Wang et al. | Oct 2011 | B2 |
| 8045153 | Mimura et al. | Oct 2011 | B2 |
| 8045263 | Yaroslavsky et al. | Oct 2011 | B2 |
| 8050735 | Feke et al. | Nov 2011 | B2 |
| 8055035 | Okugawa et al. | Nov 2011 | B2 |
| 8078243 | Ediger et al. | Dec 2011 | B2 |
| 8078263 | Zeman et al. | Dec 2011 | B2 |
| 8078264 | Basilion | Dec 2011 | B2 |
| 8078265 | Mahmood et al. | Dec 2011 | B2 |
| 8082024 | Alfano et al. | Dec 2011 | B2 |
| 8107696 | Pote et al. | Jan 2012 | B2 |
| 8108022 | Balberg et al. | Jan 2012 | B2 |
| 8118206 | Zand et al. | Feb 2012 | B2 |
| 8121671 | Hull et al. | Feb 2012 | B2 |
| 8125623 | Munger et al. | Feb 2012 | B2 |
| 8129105 | Zuckerman | Mar 2012 | B2 |
| 8131332 | Maynard et al. | Mar 2012 | B2 |
| 8131349 | Okawa et al. | Mar 2012 | B2 |
| 8135449 | Wilson et al. | Mar 2012 | B2 |
| 8139211 | Yaroslavsky et al. | Mar 2012 | B2 |
| 8140147 | Maynard et al. | Mar 2012 | B2 |
| 8145295 | Boyden et al. | Mar 2012 | B2 |
| 8149418 | Tearney et al. | Apr 2012 | B2 |
| 8157807 | Ferren et al. | Apr 2012 | B2 |
| 8158919 | Maeda et al. | Apr 2012 | B2 |
| 8160680 | Boyden et al. | Apr 2012 | B2 |
| 8180421 | Phillips et al. | May 2012 | B2 |
| 8180436 | Boyden et al. | May 2012 | B2 |
| 8185180 | Diab et al. | May 2012 | B2 |
| 8188446 | Ohno | May 2012 | B2 |
| 8189201 | Haisch et al. | May 2012 | B2 |
| 8189887 | Kollias et al. | May 2012 | B2 |
| 8190231 | Miwa et al. | May 2012 | B2 |
| 8190242 | Demos et al. | May 2012 | B2 |
| 8204579 | Nielsen et al. | Jun 2012 | B2 |
| 8213005 | Masilamani et al. | Jul 2012 | B2 |
| 8218143 | Gupta | Jul 2012 | B2 |
| 8224427 | Kopriva | Jul 2012 | B2 |
| 8227766 | Chapman | Jul 2012 | B2 |
| 8228368 | Zhao et al. | Jul 2012 | B2 |
| 8229548 | Frangioni | Jul 2012 | B2 |
| 8231526 | Yabe et al. | Jul 2012 | B2 |
| 8238993 | Maynard et al. | Aug 2012 | B2 |
| 8243269 | Matousek et al. | Aug 2012 | B2 |
| 8246611 | Paithankar et al. | Aug 2012 | B2 |
| 8255040 | Goldman et al. | Aug 2012 | B2 |
| 8260010 | Chhibber et al. | Sep 2012 | B2 |
| 8270689 | Liang et al. | Sep 2012 | B2 |
| 8279275 | Gono et al. | Oct 2012 | B2 |
| 8280140 | Levenson et al. | Oct 2012 | B2 |
| 8280471 | Rainone et al. | Oct 2012 | B2 |
| 8285015 | Demos | Oct 2012 | B2 |
| 8285353 | Choi et al. | Oct 2012 | B2 |
| 8289522 | Tearney et al. | Oct 2012 | B2 |
| 8294081 | Rosenheimer et al. | Oct 2012 | B2 |
| 8295901 | Tobola et al. | Oct 2012 | B2 |
| 8311607 | Zelenchuk | Nov 2012 | B2 |
| 8320650 | Demos et al. | Nov 2012 | B2 |
| 8326404 | Zeng et al. | Dec 2012 | B2 |
| 8326406 | Ntziachristos et al. | Dec 2012 | B2 |
| 8330087 | Domenicali | Dec 2012 | B2 |
| 8332006 | Naganuma et al. | Dec 2012 | B2 |
| 8335550 | Segman | Dec 2012 | B2 |
| 8346329 | Xu et al. | Jan 2013 | B2 |
| 8351026 | Stern | Jan 2013 | B2 |
| 8351041 | Leveque et al. | Jan 2013 | B2 |
| 8358821 | Yamaguchi et al. | Jan 2013 | B2 |
| 8380268 | Georgakoudi et al. | Feb 2013 | B2 |
| 8385615 | Levenson et al. | Feb 2013 | B2 |
| 8391961 | Levenson et al. | Mar 2013 | B2 |
| 8403862 | Grinvald et al. | Mar 2013 | B2 |
| 8417324 | Mycek et al. | Apr 2013 | B2 |
| 8423127 | Mahmood et al. | Apr 2013 | B2 |
| 8452357 | Rebec et al. | May 2013 | B2 |
| 8452384 | Ince | May 2013 | B2 |
| 8463006 | Prokoski | Jun 2013 | B2 |
| 8467857 | Kim et al. | Jun 2013 | B2 |
| 8491120 | Kahn et al. | Jul 2013 | B2 |
| 8496695 | Kang et al. | Jul 2013 | B2 |
| 8498682 | Markle et al. | Jul 2013 | B2 |
| 8504140 | Feke et al. | Aug 2013 | B2 |
| 8521261 | Okawa et al. | Aug 2013 | B2 |
| 8538195 | Robinson | Sep 2013 | B2 |
| 8538504 | Kleen et al. | Sep 2013 | B2 |
| 8540393 | Mizuno | Sep 2013 | B2 |
| 8543180 | Bechtel et al. | Sep 2013 | B2 |
| 8547425 | Ishihara | Oct 2013 | B2 |
| 8562657 | Ferren et al. | Oct 2013 | B2 |
| 8574859 | Lin et al. | Nov 2013 | B2 |
| 8581970 | Yamazaki et al. | Nov 2013 | B2 |
| 8588893 | Jaeb et al. | Nov 2013 | B2 |
| 8593513 | Yamaguchi et al. | Nov 2013 | B2 |
| 8598540 | Moy et al. | Dec 2013 | B2 |
| 8605974 | Liang et al. | Dec 2013 | B2 |
| 8609358 | Sebastian et al. | Dec 2013 | B2 |
| 8617057 | Morishita et al. | Dec 2013 | B2 |
| 8619257 | Plowman et al. | Dec 2013 | B2 |
| 8620411 | Stamatas et al. | Dec 2013 | B2 |
| 8626271 | Dunki-Jacobs et al. | Jan 2014 | B2 |
| 8634607 | Levenson et al. | Jan 2014 | B2 |
| 8639043 | Levenson et al. | Jan 2014 | B2 |
| 8644663 | Viellerobe et al. | Feb 2014 | B2 |
| 8644900 | Balberg et al. | Feb 2014 | B2 |
| 8644911 | Panasyuk et al. | Feb 2014 | B1 |
| 8660637 | Crowley | Feb 2014 | B2 |
| 8676283 | Matter et al. | Mar 2014 | B2 |
| 8676302 | Wang et al. | Mar 2014 | B2 |
| 8705042 | Haisch et al. | Apr 2014 | B2 |
| 8718737 | Diab et al. | May 2014 | B2 |
| 9042967 | Dacosta et al. | May 2015 | B2 |
| 9675238 | Iida | Jun 2017 | B2 |
| 9696897 | Garcia | Jul 2017 | B2 |
| 10200625 | Marcelpoil et al. | Feb 2019 | B2 |
| 10438356 | Dacosta | Oct 2019 | B2 |
| 11154198 | Dacosta et al. | Oct 2021 | B2 |
| 11266345 | Saiko et al. | Mar 2022 | B2 |
| 20010034477 | Mansfield et al. | Oct 2001 | A1 |
| 20010047136 | Domanik et al. | Nov 2001 | A1 |
| 20020058864 | Mansfield et al. | May 2002 | A1 |
| 20020091324 | Kollias et al. | Jul 2002 | A1 |
| 20020138008 | Tsujita et al. | Sep 2002 | A1 |
| 20020146160 | Parker et al. | Oct 2002 | A1 |
| 20030001104 | Sendai et al. | Jan 2003 | A1 |
| 20030049175 | Buechler et al. | Mar 2003 | A1 |
| 20030055341 | Banerjee | Mar 2003 | A1 |
| 20030068274 | Jungmann et al. | Apr 2003 | A1 |
| 20030082105 | Fischman et al. | May 2003 | A1 |
| 20030109787 | Black | Jun 2003 | A1 |
| 20030113934 | Kwon | Jun 2003 | A1 |
| 20030123056 | Barnes et al. | Jul 2003 | A1 |
| 20030135122 | Bambot et al. | Jul 2003 | A1 |
| 20030158470 | Wolters et al. | Aug 2003 | A1 |
| 20030160182 | Petrich et al. | Aug 2003 | A1 |
| 20030195401 | Tian et al. | Oct 2003 | A1 |
| 20030206301 | Cline et al. | Nov 2003 | A1 |
| 20030216626 | Tsujita et al. | Nov 2003 | A1 |
| 20040006276 | Demos et al. | Jan 2004 | A1 |
| 20040034292 | Mansfield et al. | Feb 2004 | A1 |
| 20040064053 | Chang et al. | Apr 2004 | A1 |
| 20040073120 | Motz et al. | Apr 2004 | A1 |
| 20040077951 | Lin et al. | Apr 2004 | A1 |
| 20040101826 | Jones et al. | May 2004 | A1 |
| 20040143190 | Schnitzer | Jul 2004 | A1 |
| 20040147843 | Bambot et al. | Jul 2004 | A1 |
| 20040148141 | Tsujita et al. | Jul 2004 | A1 |
| 20040186382 | Modell et al. | Sep 2004 | A1 |
| 20040186383 | Rava et al. | Sep 2004 | A1 |
| 20040196463 | Cline et al. | Oct 2004 | A1 |
| 20040225222 | Zeng et al. | Nov 2004 | A1 |
| 20040236229 | Freeman et al. | Nov 2004 | A1 |
| 20040260365 | Groseth et al. | Dec 2004 | A1 |
| 20050021235 | Bar-Or et al. | Jan 2005 | A1 |
| 20050064602 | Kaufman et al. | Mar 2005 | A1 |
| 20050119548 | Lin et al. | Jun 2005 | A1 |
| 20050131304 | Stamatas et al. | Jun 2005 | A1 |
| 20050143662 | Marchitto et al. | Jun 2005 | A1 |
| 20060004292 | Beylin | Jan 2006 | A1 |
| 20060008866 | Flick et al. | Jan 2006 | A1 |
| 20060013454 | Flewelling et al. | Jan 2006 | A1 |
| 20060030761 | Raskas | Feb 2006 | A1 |
| 20060077581 | Schwiegerling et al. | Apr 2006 | A1 |
| 20060082768 | Wilson et al. | Apr 2006 | A1 |
| 20060089556 | Bambot et al. | Apr 2006 | A1 |
| 20060135869 | Farina | Jun 2006 | A1 |
| 20060151709 | Hahl | Jul 2006 | A1 |
| 20060188389 | Levy | Aug 2006 | A1 |
| 20060241495 | Kurtz | Oct 2006 | A1 |
| 20060241499 | Irion et al. | Oct 2006 | A1 |
| 20060249690 | Pfister et al. | Nov 2006 | A1 |
| 20060253261 | Maier et al. | Nov 2006 | A1 |
| 20060270919 | Brenner | Nov 2006 | A1 |
| 20070004972 | Cole et al. | Jan 2007 | A1 |
| 20070015963 | Fengler et al. | Jan 2007 | A1 |
| 20070024946 | Panasyuk et al. | Feb 2007 | A1 |
| 20070038124 | Fulghum et al. | Feb 2007 | A1 |
| 20070038126 | Pyle et al. | Feb 2007 | A1 |
| 20070049809 | Bechtel et al. | Mar 2007 | A1 |
| 20070060804 | Thompson et al. | Mar 2007 | A1 |
| 20070073156 | Zilberman et al. | Mar 2007 | A1 |
| 20070073158 | Sendai | Mar 2007 | A1 |
| 20070080305 | Maitrejean et al. | Apr 2007 | A1 |
| 20070093700 | Wang et al. | Apr 2007 | A1 |
| 20070093703 | Sievert et al. | Apr 2007 | A1 |
| 20070129613 | Rochester et al. | Jun 2007 | A1 |
| 20070129615 | Backman et al. | Jun 2007 | A1 |
| 20070135873 | Johansson et al. | Jun 2007 | A1 |
| 20070149868 | Blank et al. | Jun 2007 | A1 |
| 20070149882 | Wedel | Jun 2007 | A1 |
| 20070156036 | Pilon et al. | Jul 2007 | A1 |
| 20070167836 | Scepanovic et al. | Jul 2007 | A1 |
| 20070167838 | Hubble et al. | Jul 2007 | A1 |
| 20070179366 | Pewzner et al. | Aug 2007 | A1 |
| 20070212038 | Asai et al. | Sep 2007 | A1 |
| 20070213618 | Li et al. | Sep 2007 | A1 |
| 20070223797 | Kaneko | Sep 2007 | A1 |
| 20070239031 | Lee et al. | Oct 2007 | A1 |
| 20070239034 | Knoche et al. | Oct 2007 | A1 |
| 20070244395 | Wang et al. | Oct 2007 | A1 |
| 20070276199 | Ediger et al. | Nov 2007 | A1 |
| 20080013166 | Haisch et al. | Jan 2008 | A1 |
| 20080013960 | Tearney et al. | Jan 2008 | A1 |
| 20080014463 | Varadarajan et al. | Jan 2008 | A1 |
| 20080045799 | Whitehead et al. | Feb 2008 | A1 |
| 20080051664 | Demos et al. | Feb 2008 | A1 |
| 20080051665 | Xu et al. | Feb 2008 | A1 |
| 20080058587 | Boyden et al. | Mar 2008 | A1 |
| 20080058785 | Boyden et al. | Mar 2008 | A1 |
| 20080058795 | Boyden et al. | Mar 2008 | A1 |
| 20080059070 | Boyden | Mar 2008 | A1 |
| 20080076985 | Matousek et al. | Mar 2008 | A1 |
| 20080082000 | Thoms | Apr 2008 | A1 |
| 20080086038 | Thornton | Apr 2008 | A1 |
| 20080091110 | Zelenchuk | Apr 2008 | A1 |
| 20080097225 | Tearney et al. | Apr 2008 | A1 |
| 20080103355 | Boyden et al. | May 2008 | A1 |
| 20080103375 | Kiani | May 2008 | A1 |
| 20080103384 | Pfister | May 2008 | A1 |
| 20080119832 | Cronin | May 2008 | A1 |
| 20080128505 | Challa et al. | Jun 2008 | A1 |
| 20080132793 | Kollias et al. | Jun 2008 | A1 |
| 20080154102 | Frangioni et al. | Jun 2008 | A1 |
| 20080161699 | Zeng et al. | Jul 2008 | A1 |
| 20080177140 | Cline et al. | Jul 2008 | A1 |
| 20080183059 | LePlante et al. | Jul 2008 | A1 |
| 20080188727 | Benaron et al. | Aug 2008 | A1 |
| 20080188736 | Bambot et al. | Aug 2008 | A1 |
| 20080194968 | Drugge | Aug 2008 | A1 |
| 20080221416 | Baker | Sep 2008 | A1 |
| 20080228037 | Cline et al. | Sep 2008 | A1 |
| 20080228049 | Black | Sep 2008 | A1 |
| 20080249381 | Muller et al. | Oct 2008 | A1 |
| 20080269617 | Kohler et al. | Oct 2008 | A1 |
| 20080300493 | Gatto et al. | Dec 2008 | A1 |
| 20080312540 | Ntziachristos | Dec 2008 | A1 |
| 20090018418 | Markle et al. | Jan 2009 | A1 |
| 20090043296 | Foster et al. | Feb 2009 | A1 |
| 20090060304 | Gulfo et al. | Mar 2009 | A1 |
| 20090073439 | Tearney et al. | Mar 2009 | A1 |
| 20090075391 | Fulghum, Jr. | Mar 2009 | A1 |
| 20090118600 | Ortiz et al. | May 2009 | A1 |
| 20090118622 | Durkin et al. | May 2009 | A1 |
| 20090124854 | Yamaguchi et al. | May 2009 | A1 |
| 20090131800 | Liang | May 2009 | A1 |
| 20090137908 | Patwardhan | May 2009 | A1 |
| 20090137952 | Ramamurthy et al. | May 2009 | A1 |
| 20090163819 | De Kok et al. | Jun 2009 | A1 |
| 20090187108 | Tang et al. | Jul 2009 | A1 |
| 20090192349 | Berguer et al. | Jul 2009 | A1 |
| 20090209866 | Abovitz et al. | Aug 2009 | A1 |
| 20090234234 | Machida | Sep 2009 | A1 |
| 20090236541 | Lomnes et al. | Sep 2009 | A1 |
| 20090247881 | Maeda et al. | Oct 2009 | A1 |
| 20090249500 | Zhao et al. | Oct 2009 | A1 |
| 20090264772 | Van Der Brug et al. | Oct 2009 | A1 |
| 20090268011 | Scott et al. | Oct 2009 | A1 |
| 20090270702 | Zeng et al. | Oct 2009 | A1 |
| 20090289200 | Ishii | Nov 2009 | A1 |
| 20090299196 | Bawendi et al. | Dec 2009 | A1 |
| 20100010340 | Godavarty et al. | Jan 2010 | A1 |
| 20100016688 | Debreczeny et al. | Jan 2010 | A1 |
| 20100041998 | Postel | Feb 2010 | A1 |
| 20100069758 | Barnes et al. | Mar 2010 | A1 |
| 20100140461 | Sprigle et al. | Jun 2010 | A1 |
| 20100160752 | Chance | Jun 2010 | A1 |
| 20100168586 | Hillman et al. | Jul 2010 | A1 |
| 20100174160 | Chance | Jul 2010 | A1 |
| 20100185099 | Johansson et al. | Jul 2010 | A1 |
| 20100217129 | El-Deiry et al. | Aug 2010 | A1 |
| 20100234739 | Nakaoka et al. | Sep 2010 | A1 |
| 20100234740 | Roessler et al. | Sep 2010 | A1 |
| 20100241055 | Dacey, Jr. et al. | Sep 2010 | A1 |
| 20100256469 | Cook et al. | Oct 2010 | A1 |
| 20100256504 | Moreau-Gaudry et al. | Oct 2010 | A1 |
| 20100268091 | Takaoka | Oct 2010 | A1 |
| 20100331705 | Het Hooft et al. | Dec 2010 | A1 |
| 20100331706 | Hasegawa | Dec 2010 | A1 |
| 20100331707 | Fukutani et al. | Dec 2010 | A1 |
| 20110015591 | Hanson et al. | Jan 2011 | A1 |
| 20110042580 | Wilson et al. | Feb 2011 | A1 |
| 20110063427 | Fengler et al. | Mar 2011 | A1 |
| 20110085714 | Yan et al. | Apr 2011 | A1 |
| 20110087111 | Ntziachristos | Apr 2011 | A1 |
| 20110098575 | Stamnes et al. | Apr 2011 | A1 |
| 20110102566 | Zakian et al. | May 2011 | A1 |
| 20110117025 | Dacosta et al. | May 2011 | A1 |
| 20110124989 | Edgar et al. | May 2011 | A1 |
| 20110144504 | Tearney et al. | Jun 2011 | A1 |
| 20110184260 | Robinson et al. | Jul 2011 | A1 |
| 20110201940 | Wang et al. | Aug 2011 | A1 |
| 20110213252 | Fulghum | Sep 2011 | A1 |
| 20110224553 | Stothers et al. | Sep 2011 | A1 |
| 20110237909 | Black | Sep 2011 | A1 |
| 20110275899 | Tearney et al. | Nov 2011 | A1 |
| 20110275900 | Gilhuly et al. | Nov 2011 | A1 |
| 20110295125 | Lin et al. | Dec 2011 | A1 |
| 20110313296 | Johnson et al. | Dec 2011 | A9 |
| 20120004508 | McDowall et al. | Jan 2012 | A1 |
| 20120004557 | McDowall et al. | Jan 2012 | A1 |
| 20120016230 | Kishima et al. | Jan 2012 | A1 |
| 20120029348 | Yaroslavsky et al. | Feb 2012 | A1 |
| 20120053429 | Trepagnier et al. | Mar 2012 | A1 |
| 20120059254 | Lifan et al. | Mar 2012 | A1 |
| 20120065484 | Hull et al. | Mar 2012 | A1 |
| 20120071764 | Yaroslavsky et al. | Mar 2012 | A1 |
| 20120078075 | Maynard et al. | Mar 2012 | A1 |
| 20120089031 | Ince | Apr 2012 | A1 |
| 20120108982 | Hoyt et al. | May 2012 | A1 |
| 20120197096 | Ridder et al. | Aug 2012 | A1 |
| 20120197134 | Okawa et al. | Aug 2012 | A1 |
| 20120220880 | Zuluaga | Aug 2012 | A1 |
| 20120226167 | Zuluaga | Sep 2012 | A1 |
| 20120265078 | Goldman et al. | Oct 2012 | A1 |
| 20120277558 | Barber et al. | Nov 2012 | A1 |
| 20120283531 | Maynard et al. | Nov 2012 | A1 |
| 20120302892 | Lue et al. | Nov 2012 | A1 |
| 20130018236 | Altshuler et al. | Jan 2013 | A1 |
| 20130033589 | Demos | Feb 2013 | A1 |
| 20130066215 | Tearney et al. | Mar 2013 | A1 |
| 20130072769 | Zuckerman | Mar 2013 | A1 |
| 20130100455 | Tearney et al. | Apr 2013 | A1 |
| 20130131488 | Zeng et al. | May 2013 | A1 |
| 20130148106 | Tearney et al. | Jun 2013 | A1 |
| 20130217985 | Dvorsky et al. | Aug 2013 | A1 |
| 20130237860 | Ince | Sep 2013 | A1 |
| 20130302746 | Liang et al. | Nov 2013 | A1 |
| 20140031647 | Lin et al. | Jan 2014 | A1 |
| 20140050667 | Wang et al. | Feb 2014 | A1 |
| 20140055605 | Moy et al. | Feb 2014 | A1 |
| 20140058220 | LeBoeuf et al. | Feb 2014 | A1 |
| 20140073885 | Frangioni | Mar 2014 | A1 |
| 20140128730 | Wang et al. | May 2014 | A1 |
| 20140207003 | Gilhuly | Jul 2014 | A1 |
| 20140221843 | You et al. | Aug 2014 | A1 |
| 20140300891 | Alfano et al. | Oct 2014 | A1 |
| 20150042877 | O'Neill et al. | Feb 2015 | A1 |
| 20150145966 | Krieger et al. | May 2015 | A1 |
| 20150182196 | Ninomiya et al. | Jul 2015 | A1 |
| 20150192521 | Lausch | Jul 2015 | A1 |
| 20150216418 | Ammon | Aug 2015 | A1 |
| 20160045114 | Dacosta et al. | Feb 2016 | A1 |
| 20160187199 | Brunk et al. | Jun 2016 | A1 |
| 20160289729 | Richards | Oct 2016 | A1 |
| 20160324506 | Tariyal | Nov 2016 | A1 |
| 20170023599 | Richards | Jan 2017 | A1 |
| 20170183705 | Hicks | Jun 2017 | A1 |
| 20170236281 | Dacosta | Aug 2017 | A1 |
| 20180188108 | Fawzy | Jul 2018 | A1 |
| 20180242848 | Dacosta et al. | Aug 2018 | A1 |
| 20180325377 | Dacosta et al. | Nov 2018 | A1 |
| 20190216326 | Cross et al. | Jul 2019 | A1 |
| 20200104998 | Dacosta | Apr 2020 | A1 |
| 20210259552 | Dacosta et al. | Aug 2021 | A1 |
| Number | Date | Country |
|---|---|---|
| 2360229 | May 2007 | CA |
| 2231799 | Jul 2007 | CA |
| 2306795 | Jan 2009 | CA |
| 2343401 | Jan 2009 | CA |
| 2305721 | Feb 2009 | CA |
| 2190374 | Jul 2010 | CA |
| 2371886 | Jan 2012 | CA |
| 2544204 | Jul 2013 | CA |
| 2331090 | Oct 2013 | CA |
| 2533621 | Dec 2013 | CA |
| 2489915 | Jan 2014 | CA |
| 2501098 | Apr 2014 | CA |
| 2685000 | Apr 2014 | CA |
| 1623001 | Jun 2005 | CN |
| 1652012 | Aug 2005 | CN |
| 101943661 | Jan 2011 | CN |
| 102099671 | Jun 2011 | CN |
| 694165 | Mar 1998 | EP |
| 1071473 | Jan 2001 | EP |
| 930916 | Sep 2001 | EP |
| 864864 | Jan 2003 | EP |
| 850018 | Apr 2003 | EP |
| 779508 | Jun 2003 | EP |
| 1239771 | Oct 2004 | EP |
| 1089067 | Dec 2004 | EP |
| 783867 | Feb 2006 | EP |
| 1026999 | Jun 2006 | EP |
| 1161924 | Dec 2006 | EP |
| 1519769 | Feb 2007 | EP |
| 765134 | Jul 2007 | EP |
| 1692510 | Jan 2008 | EP |
| 1144049 | Mar 2008 | EP |
| 1383542 | Apr 2008 | EP |
| 1912059 | Apr 2008 | EP |
| 1314395 | Jun 2008 | EP |
| 1520508 | Nov 2008 | EP |
| 1304955 | Dec 2008 | EP |
| 1281405 | Jan 2009 | EP |
| 1217943 | Apr 2009 | EP |
| 2073706 | Jul 2009 | EP |
| 1196088 | Sep 2009 | EP |
| 1778076 | Sep 2009 | EP |
| 1277436 | Dec 2009 | EP |
| 1830705 | Dec 2010 | EP |
| 1617761 | Jan 2012 | EP |
| 1045717 | Mar 2012 | EP |
| 2291640 | Dec 2012 | EP |
| 1576181 | Aug 2013 | EP |
| 3171765 | May 2018 | EP |
| 2367125 | Mar 2002 | GB |
| S59-40830 | Mar 1984 | JP |
| H02-195235 | Aug 1990 | JP |
| H04-127039 | Apr 1992 | JP |
| H05337142 | Dec 1993 | JP |
| H08-320321 | Dec 1996 | JP |
| H09-173048 | Jul 1997 | JP |
| 10096697 | Apr 1998 | JP |
| 10328129 | Dec 1998 | JP |
| 2001503645 | Mar 2001 | JP |
| 2004127039 | Apr 2004 | JP |
| 2004237081 | Aug 2004 | JP |
| 2005233636 | Sep 2005 | JP |
| 2005331889 | Dec 2005 | JP |
| 2006081842 | Mar 2006 | JP |
| 2006122335 | May 2006 | JP |
| 2006515065 | May 2006 | JP |
| 2006187598 | Jul 2006 | JP |
| 2007050010 | Mar 2007 | JP |
| 2007151782 | Jun 2007 | JP |
| 2007515947 | Jun 2007 | JP |
| 2007516009 | Jun 2007 | JP |
| 2007198845 | Aug 2007 | JP |
| 2007524389 | Aug 2007 | JP |
| 2007526478 | Sep 2007 | JP |
| 4475923 | Jun 2010 | JP |
| 2011103118 | May 2011 | JP |
| 2011521237 | Jul 2011 | JP |
| 02061405 | Aug 2002 | NO |
| 1994021816 | Sep 1994 | WO |
| 9715226 | May 1997 | WO |
| 200025114 | May 2000 | WO |
| 2002055729 | Jul 2002 | WO |
| 2004025556 | Mar 2004 | WO |
| 2006007715 | Jan 2006 | WO |
| 2006101736 | Sep 2006 | WO |
| 2007035829 | Mar 2007 | WO |
| 2008028298 | Mar 2008 | WO |
| 2009140757 | Nov 2009 | WO |
| 2012075028 | Jun 2012 | WO |
| 2012083349 | Jun 2012 | WO |
| 2016011534 | Jan 2016 | WO |
| 2017079844 | May 2017 | WO |
| 2019092509 | May 2019 | WO |
| Entry |
|---|
| Notice of Allowance dated Jan. 25, 2023 in U.S. Appl. No. 16/593,174. |
| Corrected Notice of Allowability dated Feb. 1, 2023 in U.S. Appl. No. 16/593,174. |
| Final Office Action dated Jan. 31, 2023 in Japanese Patent Application No. 2020-159809. |
| Office Action dated Mar. 1, 2023 in U.S. Appl. No. 17/193,318. |
| International Search Report in PCT/CA2015/000444 dated Oct. 30, 2015. |
| Office Action in CN Appln No. 2015-10283523.6 dated Apr. 14, 2017. |
| Office Action in JP Appln No. 2014-207852 dated Oct. 27, 2015. |
| Notice of Grounds of Rejection in JP Appln No. 2011-509826 dated Oct. 8, 2013. |
| Office Action in CA Appln No. 2,724,973 dated Jun. 25, 2014. |
| Communication in EP Appln No. 09749361.3 dated Nov. 6, 2012. |
| Extended European Search Report in PCT/CA2009/000680 dated Nov. 6, 2012. |
| Notification of Reexamination in CN Appln No. 200980128426.2 dated Jul. 8, 2014. |
| Extended Supplementary European Search Report in EP Appln No. 09 74 9361, dated Oct. 25, 2012. |
| Non-Final Office Action in U.S. Appl. No. 15/328,214, dated Apr. 3, 2018. |
| Non-Final Office Action in U.S. Appl. No. 12/992,040, dated Jun. 25, 2014. |
| Final Office Action in U.S. Appl. No. 12/992,040, dated Dec. 29, 2014. |
| Notice of Allowance in U.S. Appl. No. 12/992,040, dated Jan. 23, 2015. |
| Notification of Second Office Action in CN Appln No. 200980128426.2, dated Dec. 24, 2012. |
| Treuillet, S. et al. “Three-Dimensional Assessment of Skin Wounds Using a Standard Digital Camera”, IEEE Transactions on Medical Imaging, May 2009, vol. 28, No. 5, pp. 752-762. |
| JP Office Action in JP Application No. 2011/509825 dated Jun. 3, 2014. |
| Falanga et al., “Wounding of Bioengineered Skin: Cellular and Molecular Aspects After Injury,” J. Invest Dermatol 2002, 119(3):653-660 (8 pages). |
| Cutting et al., Journal of Wound Care 1994, 3:198-201. |
| Dow G. In: Krasner et al. eds. “Chronic Wound Care: A Clinical Source Book for Healthcare Professionals,” 3rd ed. Wayne Pa: HMP Communications 2001:343-356. |
| “Physiological basis of wound healing in Developments in wound care,” PJB Publications Ltd., 5-17, 1994. |
| Cooper et al., “Wound Infection and Microbiology.” Medical Communications (UK) Ltd for Johnson & Johnson Medical, 2003. |
| Mortimer PS. In: Doyle et al. editors. Oxford Textbook of Palliative Medicine (2nd ed). Oxford: Oxford University Press, 1998, 617-627. |
| Englund F. RCN Contact 1993. |
| Galpin et al., “Sepsis Associated with Decubitus Ulcers,” The American Journal of Medicine, Sep. 1976, vol. 61 pp. 346-350 (5 pages). |
| Ayton M., “Wound Care: wounds that won't heal,” Nurs Times Nov. 1985; 81(46): suppl 16-19 (1 page). |
| Grocott P., “The palliative management of fungating malignant wounds.”, J Wound Care May 1995, 4(5);240-242 (1 page). |
| Collier M., The assessment of patients with malignant fungating wounds-a holistic approach: Part 1., Nurs Times Oct. 29-Nov. 4, 1997; 93(44): suppl 1-4 (1 page). |
| Grocott P., “The management of fungating wounds.” J Wound Care May 1999; 8(5):232-234 (1 page). |
| Young T., “The challenge of managing fungating wounds.” Oct. 1997; 3(9): 41-44 (1 page). |
| Website http://www.iec.ch/online news/etech/arch_2006/etech_0906/focus.htm. |
| Chinese Office Action for Chinese Patent Application No. CN100098 dated Feb. 1, 2012 (6 pages). |
| Zhu, “Thesis: Non-invasive Optical Technologies to Monitor Wound Healing”, Drexel University, Dec. 2007. |
| Andersson-Engels, et al. “In vivo fluorescence imaging for tissue diagnostics”, Phys. Med. Biol. 42 (1997):815-824 (10 pages). |
| International Search Report dated Sep. 24, 2009 for International Application No. PCT/CA2009/000680 (3 pages). |
| Broer et al. “Laser-induced Fluorescence Spectroscopy for Real-Time Tissue Differentiation”, Medical Laser Application 19:45-53, 2004 (9 pages). |
| DaCosta, et al., “Autofluorescence characterisation of isolated whole crypts and primary cultured human epithelial cells from normal, hyperplastic, and adenomatous colonic mucosa”, J Clin Pathol 2005; 58:766-774 (10 pages). |
| Jacques et al., “PDT with ALA/PPIX is enhanced by prolonged light exposure putatively by targeting mitochondria.” SPIE Proceedings, vol. 2972, Optical Methods for Tumor Treatment and Detection, ed T. Dougherty, San Jose, Feb. 1997 (5 pages). |
| Bowler et al., “Wound Microbiology and Associated Approaches to Wound Management,” Clinical Microbiology Reviews 2001, 14:244-269 (27 pages). |
| Dow et al., “Infection in chronic wounds: controversies in diagnosis and treatment.”, Ostomy/VVound Management 1999, 45:23-40 (Abstract—1 page). |
| Winter, “Formation of the Scab and the Rate of Epithelization of Superficial Wounds in the Skin of the Young Domestic Pig,” Nature, Jan. 20, 1962, 193:293-294 (2 pages). |
| Perednia, “What dermatologists should know about digital imaging,” J Am Acad Dermatol 1991, 25:89-108 (20 pages). |
| Serena et al., “The Lack of Reliability of Clinical Examination in the Diagnosis of Wound Infection: Preliminary Communication,” The International Journal of Lower Extremity Wounds, V7(1); Mar. 2008:32-35 (4 pages). |
| Gardner et al., “Diagnostic Validity of Semiquantitative Swab Cultures,” Wounds Feb. 2007; vol. 19, Issue 2:31-38 (8 pages). |
| Falanga et al., “Workshop on the Pathogenesis of Chronic Wounds,” J. Invest Dermatol 1994, 102(1):125-127 (4 pages). |
| Kingsley et al., “A proactive approach to wound infection”, Nursing Standard Apr. 2001; 15(30): 50-54, 56 & 58 (6 pages). |
| DaCosta et al., “Molecular Fluorescence Excitation-Emission Matrices Relevant to Tissue Spectroscopy”, Photochemistry and Photobiology Oct. 2003, 78(4):384-392 (9 pages). |
| DaCosta et al., “New optical technologies for earlier endoscopic diagnosis of premalignant gastrointestinal lesions”, Journal of Gastroenterology and Hepatology (2002) 17 (Suppl.) S85-S104 (20 pages). |
| Poh et al., “Direct Fluorescence Visualization of Clinically Occult High-Risk Oral Premalignant Disease Using a Simple Hand-Held Device”, Head & Neck—DOI, Jan. 2007; 29(1):71-76 (6 pages). |
| Hanibuchi et al., “Autofluorescence bronchoscopy, a novel modality for the early detection of bronchial premalignant and malignant lesions”, The Journal of Medical Investigation 2007, vol. 54:261-266 (6 pages). |
| D'Hallewin et al., “Fluorescence Detection of Bladder Cancer: A Review”, European Urology 2002, 42(5):417-425 (9 pages). |
| Yasui et al., “Determination of collagen fiber orientation in human tissue by use of polariztation measurement of molecular second-harmonic-generation light”, Applied Optics May 10, 2004; vol. 43, No. 14:2861-2867 (7 pages). |
| Dietel et al., “5-Aminolaevulinic acid (ALA) induced formation of different fluorescent porphyrins: A study of the biosynthesis of porphyrins by bacteria of the human digestive tract”, Journal of Photochemistry and Photobiology B: Biology 86:77-86, (2007) (10 pages). |
| Bissonette et al., “Current status of photodynamic therapy in dermatology.” Dermatol Clin. Jul. 1997, 15(3):507-519 (Abstract 1 page). |
| Carruth, “Clinical applications of photodynamic therapy.”, Int. J. Clin. Pract. Jan.-Feb. 1998; 52(1):39-42 (Abstract 1 page). |
| Dougherty et al. “Photodynamic Therapy”, Journal of the National Cancer Institute, vol. 90, No. 12, Jun. 17, 1998:889-905 (17 pages). |
| Jori et al., “Photodynamic Therapy in the Treatment of Microbial Infections: Basic Principles and Perspective Applications”, Lasers in Surgery and Medicine Jun. 2006, 38(5):468-481 (14 pages). |
| Hamblin et al., “Photodynamic therapy: a new antimicrobial approach to infectious disease?”, Photochem Photobiol Sci. May 2004; 3(5):436-450 (30 pages). |
| Gudgin et al., “On the role of protoporphyrin IX photoproducts in photodynamic therapy,” Journal of Photochemistry and Photobiology B: Biology 29; 1995:91-93 (3 pages). |
| Konig et al., “In vivo photoproduct formation during PDT with ALA-induced endogenous porphyrins,” J. Photochem. Photobiol. B: Biol., 18 (1993):287-290 (4 pages). |
| Georgakoudi et al., “The Mechanism of Photofrin Photobleaching and Its Consequences for Photodynamic Dosimetry,” Photochemistry and Photobiology, 1997, 65(1):135-144 (10 pages). |
| Grossweiner, “Optical Dosimetry in Photodynamic Therapy,” Lasers in Surgery and Medicine, 1986; 6:462-466 (5 pages). |
| Jongen et al., “Mathematical description of photobleaching in vivo describing the influence of tissue optics on measured fluorescence signals”, Phys. Med. Biol. 42, 1997:1701-1716 (17 pages). |
| Rhodes et al., “Iontophoretic Delivery of ALA Provides a Quantitative Model for ALA Pharmacokinetics and PpIX Phototoxicity in Human Skin,” The Society for Investigative Dermatology, Inc. 1997; 108:87-91 (6 pages). |
| Robinson et al., “Fluorescence Photobleaching of ALA-induced Protoporphyrin IX during Photodynamic Therapy of Normal Hairless Mouse Skin: The Effect of Light Dose and Irradiance and the Resulting Biological Effect,” Photochemistry and Photobiology, 1998, 67(1):140-149 (10 pages). |
| Rotomskis et al., “Spectroscopic studies of photobleaching and photoproduct formation of porphyrins used in tumor therapy,” Journal of Photochemistry and Photobiology B:Biology 33, 1996:61-67 (7 pages). |
| Grossweiner, “PDT light dosimetry revisited,” Journal of Photochemistry and Photobiology B:Biology 38, 1997:258-268 (11 pages). |
| Tonnesen et al., “Angiogenesis in Wound Healing,” The Society for Investigative Dermatology, Inc. 2000; 5(1):40-46 (7 pages). |
| Chwirot et al., “Detection of Melanomas by Digital Imaging of Spectrally Resolved Ultraviolet Light-induced Autofluorescence of Human Skin,” European Journal of Cancer, vol. 34, No. 11:1730-1734, 1998 (5 pages). |
| Bishop, “Burn wound assessment and surgical management.” Crit Care Nurs Clin North Am. 2004; 16(1):145-177 (Abstract 1 page). |
| Charles, “Radon exposure of the skin: II. Estimation of the attributable risk for skin cancer incidence,” Journal of Radiological Protection 2007, 27(3):253-274 (23 pages). |
| Pretty, “Caries detection and diagnosis: Novel technologies,” Journal of Dentistry 2006, 34(10):727-739 (13 pages). |
| Kois et al., “Detecting oral cancer: a new technique and case reports.” Dent Today, Oct. 2006; 25(10):94 and 96-97 (Abstract 1 page). |
| Bogaards et al., “Increased Brain Tumor Resection Using Fluorescence Image Guidance in a Preclinical Model,” Lasers in Surgery and Medicine 2004; 35:181-190 (10 pages). |
| Kingsley, “The Wound Infection Continuum and its Application to Clinical Practice,” Ostomy Wound Management, Jul. 2003, 49(7A Suppl):1-7 (8 pages). |
| Sibbald et al., “Increased Bacterial Burden and Infection: The Story of Nerds and Stones,” Advances in Skin & Wound Care, Oct. 2006; 19:447-461 (15 pages). |
| Bauer et al., “Angiogensis, Vasculogenesis, and Induction of Healing in Chronic Wounds,” Vascular and Endovascular Surgery vol. 39, No. 4, 2005:293-306 (14 pages). |
| Brem et al., “Cellular and molecular basis of wound healing in diabetes,” The Journal of Clinical Investigation, May 2007, vol. 117, No. 5:1219-1222 (4 pages). |
| Badiavas et al., “Treatment of Chronic Wounds With Bone Marrow-Derived Cells,” Arch Dermatol, Apr. 2003, vol. 139 (4):510-516 (7 pages). |
| Phillips, “Biologic skin substitutes.” J Dermatol Surg Oncol. Aug. 1993, 19(8):794-800 (Abstract 1 page). |
| Communication in EP Appln No. 15 824 466.5, dated Apr. 19, 2018. |
| Notice of Allowance in U.S. Appl. No. 15/328,214, dated Dec. 18, 2018. |
| Notice of Allowance in U.S. Appl. No. 15/328,214, dated Jan. 30, 2019. |
| Notice of Allowance in U.S. Appl. No. 15/328,214, dated Mar. 27, 2019. |
| Notice of Grounds of Rejection in JP Application No. 2017-503917, dated May 7, 2019. |
| First Office Action—Search Report in CN Application No. 2015800518278, dated Apr. 26, 2019. |
| Notice of Grounds of Rejection in JP Application No. 2017-503917, dated Jan. 21, 2020. |
| Notification of Second Office Action in CN Appln No. 2015800518278, dated Mar. 3, 2020. |
| Communication in EP Application No. 15824466.5, dated Apr. 3, 2020. |
| Decision of Rejection in CN Application No. 20158005182278, dated Jul. 20, 2020. |
| Song, et al., “Pork Freshness Detecting Method Based on the Change of Germ Area,” Journal of Agricultural Mechanization Research, May 31, 2009. |
| “Fluorescence video dermatoscope”—Kang Uk et al. by SOI-Korea Center of Korean Electrotechnology Research Institute (KERI), Seoul, Korean Republic. Published in J. Opt. Technol. 75 (1). |
| “Wood's lamp”—Gupta et al. by Department of Dermatology, Venereology &Leprology, Dr. S. N. Medical College, Jodhpur,India. Published in Indian J Dermatol Venereol Leprol Mar.-Apr. 2004 vol 70 Issue 2. |
| An Affordable, Portable Fluorescence Imaging Device for Skin Lesion Detection Using a Dual Wavelength Approach for Image Contrast Enhancement and Aminolaevulinic Acid-induced Protoporphyrin IX. Part I. Design, Spectral and Spatial Characteristics by Department of Chemistry and Chemical Engineering, Royal Military College of Canada. |
| Notice of Allowance in U.S. Appl. No. 16/027,775, dated Oct. 29, 2020. |
| Notice of Allowance in U.S. Appl. No. 16/027,775, dated Mar. 2, 2021. |
| Office Action in U.S. Appl. No. 14/719,493, dated Dec. 22, 2020. |
| Examination Report for IN Application No. 201717005984 dated Feb. 23, 2021. |
| U.S. Appl. No. 17/193,318, filed Mar. 5, 2021. |
| Final Office Action dated Jun. 14, 2021 in related U.S. Appl. No. 14/719,493, 28 pages. |
| Office Action dated Jun. 29, 2021 in related U.S. Appl. No. 15/965,462, 16 pages. |
| Office Action in CA Application No. 2,955,976 dated Aug. 11, 2021. |
| U.S. Appl. No. 17/407,870, filed Aug. 20, 2021. |
| U.S. Appl. No. 17/408,027, filed Aug. 20, 2021. |
| JP Office Action in JP Application No. 2020-159809 dated Sep. 28, 2021, 7 pages. |
| Office Action dated Oct. 5, 2021 in related U.S. Appl. No. 17/193,318, 24 pp. |
| Wieringa, F. P., Mastik, F., Cate, F. J., Neumann, H. A., & van der Steen, A. F. (2006). Remote non-invasive stereoscopic imaging of blood vessels: first in-vivo results of a new multispectral contrast enhancement technology. Annals of biomedical engineering, 34( 12), 1870-1878. (Year: 2006). |
| U.S. Appl. No. 17/509,914, filed Oct. 25, 2021. |
| Office Action dated Nov. 3, 2021 for U.S. Appl. No. 16/593,174, 16 PP. |
| Notice of Allowance dated Nov. 12, 2021 for U.S. Appl. No. 15/965,462, 15 pp. |
| Notice of Allowance dated Nov. 26, 2021 for U.S. Appl. No. 14/719,493. |
| Office Action dated Jan. 27, 2022 in related U.S. Appl. No. 17/193,318. |
| European Search report dated Jan. 13, 2022 for EP App No. 21187101.7. |
| Examination Report dated Mar. 10, 2022 in CN App No. 2015-800518278. |
| Office Action dated May 13, 2022 in related U.S. Appl. No. 17/193,318, 34 pages. |
| Specification and Drawings filed Jul. 1, 2022 in U.S. Appl. No. 17/856,487. |
| Office Action dated Jun. 29, 2022 in related CA application No. 2,955,976. |
| Office Action dated Jul. 19, 2022 in related JP App No. 2020-159809, 3 pages. |
| Notice of Non fully Responsive dated Aug. 2, 2022 in related U.S. Appl. No. 16/593,174. |
| Decision of Reexamination dated Aug. 24, 2022 in related CN Application No. 2015800518278, 26 pages. |
| Final Additional Submission in reply to statement filed Jun. 15, 2022 related to pre-grant opposition Indian Patent Application No. 9067/DELNP/2010, 417 pages. |
| Adiuvo Diagnostics Private Limited Vs. University Health Network, Evidence by way of an Affidavit on behalf of the opponent, 13 pages. |
| Hiram C. Polk, Jr “Early Detection of Pseudomonas Burn Infection”, Clinical Experience with Wood's Light Fluorescence, Arch Surg/vol. 98, Mar. 1969, 3 pages. |
| S Veeranna, “Wood's lamp: A modified method of examination” Indian J Dermatol Venereol Leprol Sep.-Oct. 2005 vol. 71 Issue 5, 2 pages. |
| Ev Vogeley, MD, JD, “Experience With Wood Lamp Illumination and Digital Photography in the Documentation of Bruises on Human Skin”, vol. 156, Mar. 2002, www.archpediatrics.com, 4 pages. |
| Adiuvo Diagnostics private limited Pre-grant opposition filed Jan. 3, 2021 for Pre-grant Opposition Indian patent Application: 9067/DELNP/2010, 640 pages. |
| Notice of Allowance dated May 24, 2023 in related CA Application No. 2955976. |
| Specification and Drawings filed Jun. 13, 2023 for U.S. Appl. No. 18/334,123. |
| Specification and Drawings filed Jun. 13, 2023 for U.S. Appl. No. 18/334,133. |
| Decision of Rejection dated Jul. 25, 2023 received in related JP Application No. 2020-159808. |
| Priority U.S. Appl. No. 62/584,404, filed Nov. 10, 2017. |
| Ferenczi., Web archive from the Wayback machine—“Camera review: Google turns its attention to imaging on new Nexus 5”, Digital Photography review, Dec. 3, 2013. |
| Petitioner's Opposition to Patent Owner's Motion to Amend filed Apr. 27, 2023 in U.S. Pat. No. 11,266,345. |
| Office Action dated Aug. 28, 2023 in related U.S. Appl. No. 18/334,123, 12 pages. |
| Office Action dated Oct. 23, 2023 received in related U.S. Appl. No. 18/334,133, 18 pages. |
| Number | Date | Country | |
|---|---|---|---|
| 20230245315 A1 | Aug 2023 | US |
| Number | Date | Country | |
|---|---|---|---|
| 62028386 | Jul 2014 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 16593174 | Oct 2019 | US |
| Child | 18148861 | US | |
| Parent | 15328214 | US | |
| Child | 16593174 | US |
