Detecting biological aerosols is of concern in a number of civilian and military contexts. There is growing recognition that biological agents could be employed in a terrorist attack. The most effective response to biohazards depends on detecting them as early as possible. Any delay can result in further spreading of the biological agents among the population and over a wider geographical area. Early detection will enable containment of the threat.
Detection requires characterizing biological aerosols. Characterization of biological aerosols can be performed while the aerosols are airborne, or after the biological aerosols are extracted from the air and deposited onto a solid surface (or into a liquid) for subsequent physical or chemical analysis. It would be particularly desirable to provide techniques for detecting biological aerosols while the biological aerosols are entrained in air, so that additional mechanisms are not required to extract the biological aerosols from an air stream before analysis.
Various optical methods have been used to detect biological aerosols. For example, tryptophan and nicotinamide adenine dinucleotide (NADH) are present in some concentration in most biological agents. When illuminated with light of an appropriate wavelength, tryptophan, NADH and other bio-chemical species autofluoresce with a characteristic signature. The detection of such optical signatures is thus indicative of the possible presence of biological aerosols. Prior art optical detection systems include complicated imaging optics and a plurality of discrete reflective and/or refractive components, each of which must be mounted and aligned individually, increasing the complexity and cost of such detection systems. It would be desirable to provide optical-based detection systems for biological aerosols entrained in air that have reduced part counts, reduced alignment sensitivity, and reduced assembly time. The resulting cost and time savings should enable such optical detection systems to be widely deployed if a biological agent threat is suspected.
A first aspect of the concepts disclosed herein is directed to an apparatus configured to facilitate optical detection of biological aerosols in air. The apparatus includes a first non-imaging optical component configured to direct light to be used to illuminate a sample, and a second non-imaging optical component configured to direct light from the sample to a detector configured to generate a signal indicative of the presence of biological aerosols in the sample. Significantly, in one exemplary embodiment, at least a portion of the first non-imaging optical component and at least a portion of the second non-imaging optical component are implemented as a monolithic or integrated optical structure. In a particularly preferred implementation, portions of the first non-imaging optical component and portions of the second non-imaging optical component are implemented on a first monolithic optical structure, while other portions of the first and second non-imaging optical components are implemented on a second monolithic optical structure, such that when the first and second monolithic optical structures are coupled together in a facing relationship, the first and second non-imaging optical components are completed. Preferably, at least one of the first non-imaging optical component and the second non-imaging optical component is a compound parabolic collector.
In at least one exemplary embodiment, the monolithic optical structure is fabricated using injection molding techniques. Portions of the monolithic optical structure corresponding to the first and second non-imaging optical components are preferably coated with a reflective material.
The apparatus preferably includes a light source configured to stimulate a biological aerosol to emit light, the light source being coupled with the first non-imaging optical component. In a particularly preferred embodiment, the light source is implemented using a light emitting diode (LED). The apparatus also preferably includes a detector configured to detect the light emitted from the biological aerosol, the detector being coupled with the second non-imaging optical component.
Yet another aspect of the concepts disclosed herein is directed to an apparatus for optically detecting biological aerosols in air. This apparatus includes a light source configured to stimulate a biological aerosol to emit light, a first non-imaging optical component configured to direct light away from the light source, a detector configured to detect the light emitted from the biological aerosol, and a second non-imaging optical component configured to direct light toward the detector.
Preferably, at least a portion of the first non-imaging optical component and at least a portion of the second non-imaging optical component are implemented as a monolithic structure. It is also preferred to implement at least one of the first non-imaging optical component and the second non-imaging optical component using a compound parabolic collector. The light source can be beneficially implemented as an LED.
Still another aspect of the presently disclosed novel concept is an apparatus for optically detecting biological aerosols in air. The apparatus in this implementation includes a light source configured to stimulate a biological aerosol to emit light, a detector configured to detect the light emitted from the biological aerosol, a first monolithic optical structure incorporating a plurality of first surface features, and a second monolithic optical structure incorporating a plurality of second surface features. When the first monolithic optical structure and the second monolithic optical structure are disposed in a facing relationship, the plurality of the first surface features and the plurality of second surface features define a plurality of non-imaging optical components, which include at least a first non-imaging optical component disposed adjacent to the light source and being configured to direct light to be used to stimulate the biological aerosol away from the light source, and a second non-imaging optical component configured to direct light emitted from the biological aerosol toward the detector.
Preferably, at least one of the first non-imaging optical component and the second non-imaging optical component comprises a compound parabolic collector. Each monolithic optical structure can be formed from a polymer, and each surface feature defining one of the plurality of non-imaging optical components is preferably coated with a reflective material.
The first monolithic optical structure can incorporate a plurality of third surface features, while the second monolithic optical structure can incorporate a plurality of fourth surface features, such that when the first monolithic optical structure and the second monolithic optical structure are disposed in a facing relationship, the plurality of third surface features and the plurality of fourth surface features cooperate to provide support for at least one additional component, such as at least one of a dichroic beam splitter, an emitter filter, and emission filter.
A related apparatus for optically detecting biological aerosols in air, also disclosed in detail herein, includes a first LED configured to stimulate bio-fluorescence of tryptophan, a first compound parabolic collector disposed adjacent to the first LED (the first compound parabolic collector being configured to direct light away from the first LED), a second LED configured to stimulate bio-fluorescence of nicotinamide adenine dinucleotide (NADH), a second compound parabolic collector disposed adjacent to the second LED (the second compound parabolic collector being configured to direct light away from the second LED), a first detector configured to detect bio-fluorescence associated with tryptophan, a third compound parabolic collector disposed adjacent to the first detector (the third compound parabolic collector being configured to direct light toward the first detector), a second detector configured to detect bio-fluorescence associated with NADH, and a fourth compound parabolic collector disposed adjacent to the second detector (the fourth compound parabolic collector being configured to direct light toward the second detector).
Still other embodiments of the apparatus further include a virtual impactor to separate a gaseous fluid flow in which biological particles are entrained into a major flow (that includes a minor portion of biological particles above a predetermined size), and a minor flow (that includes a major portion of the biological particles above the predetermined size). The minor flow is directed onto the substrate by means of an impactor, such that biological particles entrained in the minor flow are deposited on the substrate. It should be recognized that the incorporation of a virtual impactor is not required, and other embodiments will be implemented without a virtual impactor.
In still another embodiment, the apparatus is substantially similar to those described above, except that the apparatus includes an inlet pre-filter configured upstream of the impactor and virtual impactor (when implemented). An inlet pre-filter is a device that performs one or more of the following functions: (a) removes over-sized particles that are too large to be of interest (for example, those greater than 10 microns in diameter), (b) rejects or removes rain, snow and other precipitation, (c) restricts insects from crawling or flying into the apparatus, and (d) rejects or removes other flying debris.
Still another aspect of the inventive concept disclosed herein is directed to a method for optically detecting biological aerosols. The method comprises the steps of directing light away from a light source configured to stimulate the biological particles to emit light using a first non-imaging optical component. The light directed away from the light source is used to illuminate the biological particles, one at a time as they flow though the beam of light provided by the light source, thereby stimulating the biological particles to emit light. Light emitted from each biological particle is directed to a detector using a second non-imaging optical component.
Still other embodiments of the method further comprise the step of using a virtual impactor (disposed upstream of the optical detector described above) to separate a gaseous fluid flow in which biological particles are entrained into a major flow (that includes a minor portion of biological particles above a predetermined size) and a minor flow (that includes a major portion of the biological particles above the predetermined size.) The minor flow is directed into the optical detector.
In still another embodiment, the method is substantially similar to that described above, except that the method includes using an inlet pre-filter disposed upstream of the virtual impactor (if the virtual impactor is included), to filter the gaseous fluid before optically detecting the presence of biological material.
This Summary has been provided to introduce a few concepts in a simplified form that are further described in detail below in the Description. However, this Summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.
Various aspects and attendant advantages of one or more exemplary embodiments and modifications thereto will become more readily appreciated as the same becomes better understood by reference to the following detailed description, when taken in conjunction with the accompanying drawings, wherein:
Exemplary embodiments are illustrated in referenced Figures of the drawings. It is intended that the embodiments and Figures disclosed herein are to be considered illustrative rather than restrictive. A core feature of the concepts disclosed herein is optically detecting biological materials using non-imaging optics. While prior art optical biological detectors have used discrete optical imaging components that must be mounted and aligned individually, none have replaced imaging components with non-imaging components, nor integrated portions of the optical system into a monolithic structure. The novel approach disclosed herein opens up the opportunity to exploit high volume manufacturing methods, such as injection molding the detector. In addition, features such as filter and detector mounts can be incorporated into such integrated structures, reducing part count, component alignment sensitivity, and assembly time, all resulting in cost and time savings.
In particular, it should be recognized that while the exemplary embodiments discussed in detail below are generally referred to as biological aerosol detectors, the concept of optically detecting biological materials using non-imaging components can be applied to any sample, not simply to samples of biological aerosols. While one aspect of the concepts disclosed herein is the use of non-imaging optics for detecting the presence of biological materials in a sample of gaseous fluid (such as air), it must be recognized that the concepts disclosed herein (i.e., the use of non-imaging optics to detect the presence of biological materials) can be used to detect the presence of biological materials in other types of samples and are not limited to the detection of biological materials in a gaseous fluid.
The term “biological aerosol” as used herein is intended to refer to a sample of gaseous fluid, such as air, in which biological materials (such as biological particles) are suspended or entrained.
Light source 12 is configured to emit light having a wavelength known to induce bio-fluorescence in biological materials. Those of ordinary skill in the art will recognize that both tryptophan and NADH autofluoresce with a characteristic signature when stimulated using light of an appropriate wavelength. While many different types of light sources can be employed, particularly preferred embodiments will utilize LED light sources. LEDs emitting light in a wavelength around 280 nm can be employed for the excitation of tryptophan, while LEDs emitting light in a wavelength around 360 nm can be employed for the excitation of NADH. Further research in the excitation of tryptophan and NADH using LED light sources is likely to provide additional alternative wavelengths that may be employed for this purpose.
Non-imaging optical component 16 is configured to direct light away from light source 12, and generally toward sample volume 21, which is preferably implemented as a fluid passage configured to enable a fluid (potentially including biological material) to be continually introduced into and flowing through the sample volume (i.e., as a flow of fluid). Although the detectors disclosed herein can be configured to operate in a batch mode, wherein a fluid sample is introduced into the sample volume, analyzed, and removed to enable an additional fluid sample to be introduced into the sample volume at a later time, in an exemplary embodiment, a flow of fluid will be introduced into the sample volume at a rate selected to enable substantially continuous detection to be achieved. Those of ordinary skill in the art will readily recognize that the flow rate will be a function of the specific light sources and the characteristics of the detectors employed. While not specifically shown in
In one exemplary embodiment, the fluid is a gaseous fluid, such as air. However, the principles disclosed herein can be used with liquid samples, so long as the liquid is substantially optically transparent to the wavelengths of light emitted by the light source that are required to stimulate the fluorescence of biological materials contained within the liquid, and substantially optically transparent to the wavelengths of light emitted by the stimulated biological materials. If the liquid in the sample (or other materials contained within the sample) absorbs the light emitted by the light source and required to stimulate the fluorescence of the biological materials, or if the liquid in the sample (or other materials contained within the sample) absorbs the light emitted by the stimulated biological materials, performance of the optical biological detector will be impaired. To increase the likelihood of light from the light source reaching a biological material in the sample, it is desirable for the sample volume to have a cross-sectional size and shape that generally corresponds to an exit aperture of non-imaging optical component 16 (as well as an entrance aperture of non-imaging optical component 18).
Beam splitter 48a is configured to direct light having an appropriate first wavelength towards a CPC 50a, which in turn, directs that light of the first wavelength to a detector 14a. Similarly, beam splitter 48b is configured to direct light having an appropriate second wavelength towards a CPC 50b, which in turn, directs that light of the second wavelength to a detector 14b, and beam splitter 48c is configured to direct light having an appropriate third wavelength towards a CPC 50c, which in turn, directs that light of the third wavelength to a detector 14c. In a particularly preferred embodiment, detector 14a is configured to be responsive to wavelengths associated with the bio-fluorescence of tryptophan, detector 14b is configured to be responsive to wavelengths associated with the bio-fluorescence of NADH, and detector 14c is configured to be responsive to wavelengths of light that can be used to generate a scatter signal.
While not specifically shown, it should be recognized that non-imaging optical component 16, off-axis parabolic mirror 46, CPC 50a, CPC 50b, and CPC 50c can be implemented as one or more injection molded components, generally as described above with respect to
Again, it should be recognized that non-imaging optical component 16, CPC 50d (replacing off-axis parabolic mirror 46 of
Filters can be used to clean up the light emitted by the light source, and/or to filter out wavelengths of light not required to excite the biological materials to fluoresce. In choosing an exciter filter, historically one has had two basic choices: an interference filter or a colored glass filter. Interference filters are more expensive and require collimated light for proper operation, which can severely hamper imaging and especially non-imaging optical design approaches. Generally, UG-11 colored glass filters can be used with UV-LED light sources as an excitation filter for NADH (for wavelengths of about 340 nm to 375 nm), due to the relatively low cost of such filters. However, to date, there have not been similar glass filters available for implementation of an excited filter for the excitation of tryptophan (requiring a wavelength of about 280 nm). As a result, expensive optical interference filters are generally required for use as excitation filters for tryptophan.
One member of the family of materials potentially suitable for ultra-violet light induced fluorescence excitation of tryptophan is single crystal nickel sulfate hexahydrate. Other candidate materials include ammonium nickel sulfate hexahydrate (ANSH), potassium nickel sulfate hexahydrate (KNSH), cesium nickel sulfate hexahydrate (CNSH), and Rb2NSH (RNSH). Thus, one aspect of the present disclosure encompasses the use of such materials to achieve an excitation filter configured to stimulate the fluorescence of tryptophan. The significance of these materials can be appreciated when one considers their implementation as a filter. Instead of costly, large diameter interference filters restrictively placed in a collimated beam in the optical train, glass filters coated with such materials can be incorporated into the LED metal package, generally as discussed above. This configuration will result in a much smaller-size filter, and consequently, a comparable reduction in cost.
The embodiments discussed above are based on optically detecting the presence of biological aerosols in a sample contained in (or flowing through) a sample volume. Incorporation of additional components will enable biological aerosols to be separated from a gaseous fluid such as air, enabling a pre-concentrated sample of gaseous fluid to be directed into a sample volume for analysis (i.e., for optical detection of biological material contained therein).
Because particulates of interest, such as biological aerosols, are often present in quite small concentrations in a volume of gaseous fluid, it is highly desirable to concentrate the mass of particulates into a smaller volume of gaseous fluid. Virtual impactors can achieve such a concentration without actually removing the particulates of interest from the flow of gaseous fluid. As a result, the particulate-laden gaseous fluid flow can be passed through a series of sequentially connected virtual impactors, so that a gaseous fluid flow exiting the final virtual impactor represents a concentration of particulates 2-3 orders of magnitude greater than in the original gaseous fluid flow. Using a gaseous fluid flow more concentrated with particulates within an optical detector can potentially reduce the time required to analyze an aerosol, and/or improve the detectors' false alarm rate, and/or improve the detector's lower limit of detection.
A virtual impactor uses an aerosol's inertia to separate it from a gaseous fluid stream when the direction of the stream is turned, and a basic virtual impactor can be fabricated from a pair of opposed nozzles. Within a virtual impactor, the intake gaseous fluid coming through the inlet flows out from a nozzle directly at a second opposed nozzle into which only a “minor flow” is allowed to enter. This concept is schematically illustrated by a virtual impactor 1 shown in
As a result of inertia, most of the particulates that are greater than the selected cut size are conveyed in this small minor flow and exit the virtual impactor. Most of the particulates smaller than the virtual impactor cut size are exhausted with the majority of the inlet air, as the major flow. The stopping distance of an aerosol is an important parameter in a virtual impactor design. The cut point (size at which about 50% of the aerosols impact a surface, i.e., flow into the second nozzle) is related to the stopping distance. A 3 micron aerosol has nine times the stopping distance of a 1 micron aerosol of similar density.
For the optical biological detectors described herein, several types of virtual impactors and their variants are suitable for use in collecting samples of biological aerosols as pre-concentrated fluid streams. Because a specific design of the minor flow nozzle can be optimized for a particular size of aerosols, it is contemplated that at least some embodiments disclosed herein may include multiple nozzles, each with a different geometry, so that multiple aerosol types can be efficiently collected. In one preferred embodiment, two virtual impactors are aligned in series, such that a concentration of particulates entrained in the minor flow of gaseous fluid exiting the second virtual impactor is approximately 100 times the original concentration.
System 530 also includes a virtual impactor 532 adapted to separate the gaseous fluid into a major flow and a minor flow that includes particulates of a desired size range that are directed into a sample volume 534. A gaseous fluid is forced into virtual impactor 532 by the fan, and as described above, that gaseous fluid is separated into both a major flow and a minor flow. The major flow is directed to the exhaust, while the minor flow is directed to sample volume 534.
An optical biological aerosol detector 544 (generally consistent with those described above) is included, to analyze particulates/aerosols directed into the sample volume, to determine if the aerosols are biological in nature. Significantly, such optical detectors can detect the presence of biological particles in flow, such that a solid or liquid sample does not need to be collected, in contrast to many other types of detectors.
In many applications, it will be important that the system be able to sample a large volumetric flow of air (e.g., greater than 300 lpm). To achieve this goal, it will be important to achieve the separation of particulates from a large air volume and their concentration in a relatively smaller air volume (i.e., the minor flow). In such applications, it is contemplated that two serial in-line stages of virtual impaction may be preferable. In the first stage, 90% of the inlet gaseous fluid is discarded, and the remaining 10% of the gaseous fluid (1st stage minor flow) contains the desired aerosols. This first stage minor flow then enters a second virtual impactor stage with 90% of gaseous fluid that enters the second stage being exhausted. Therefore, the two stages have the combined effect of concentrating the outlet minor gaseous fluid volume to 1/100th of the initial inlet flow volume. This relatively small minor flow should then be in the correct range for direction into the sample volume.
Referring now to the optical biological aerosol detectors described above, it should be recognized that while specific reflector shapes have been discussed, other shapes may instead be used, including but not limited to shapes having parabolic, spherical, or flat surfaces. In addition, similar functionality can be achieved via refractive lens components. These different types of optical components (reflective and refractive) can also be used in the same system.
Various permutations of the optical layouts discussed above are to be considered to be encompassed within the scope of the novel concepts disclosed herein. For instance, the clamshell imaging component may not be included for some applications, or the conical reflector employed in some embodiments described above could be replaced with a CPC. While some embodiments discussed above measure samples in reflection, it should be recognized that other geometries are also acceptable, such as measuring samples in transmission.
Sample volume 74 is disposed between reflectors 76 and 78 (generally as described above and referred to as a clamshell reflector or clamshell imager). As described above, light from the LEDs stimulates bio-fluorescence of NADH and tryptophan naturally present in biological material. In this exemplary embodiment, sample volume 74 is configured to receive a flow of fluid substantially orthogonal to the drawing sheet. Fluorescence light from such biological materials is directed out of the clamshell reflector and generally toward an off-axis parabolic mirror 80. The fluorescence light is directed toward a beam splitter 82 (preferably implemented with a dichroic filter). Generally as described above, light of appropriate wavelengths (preferably light having wavelengths ranging from about 300 nm-400 nm) is reflected by beam splitter 82 toward an entrance aperture of a CPC 84. A detector 86 is disposed at the exit aperture of CPC 84. In a particularly preferred embodiment, detector 86 is a photomultiplier tube configured to respond to light having a wavelength of between about 300 nm and 400 nm.
Fluorescence light that passes through beam splitter 82 is then directed to a beam splitter 88 (also, for example, implemented with a dichroic filter). Light of appropriate wavelengths (preferably light having wavelengths ranging from about 400 nm-500 nm) is reflected by beam splitter 88 toward an entrance aperture of a CPC 90. A detector 92 is disposed at the exit aperture of CPC 90. In one embodiment, detector 92 is a photomultiplier tube configured to respond to light having a wavelength of between about 400 nm and 500 nm. Detectors 86 and 92 are configured to generate signals indicative of fluorescence light associated with the autofluorescence of tryptophan and NADH, and analysis of such signals can be used to determine whether a biological material is present within the fluid passing through sample volume 74.
Detector 60 also includes a laser diode 94 (preferably a laser diode emitting light having a wavelength of about 820 nm), configured to emit light that is filtered by a mask 96, and focused by a lens 98. Focused light passes through an aperture 100, and is directed to a reflector 102, which in turn directs the light through a lens 104 and an opening in off-axis parabolic mirror 72, and towards sample volume 74. Scattered light exits the sample volume and is directed towards a lens 106, a beam stop 108, and a detector 110 (preferably implemented using a photo diode). Detector 110 is configured to generate a scattering signal that can be used for several purposes. The scattering signal can be used to estimate relative sizes of particles passing through the sample volume, such that signals from detectors 86 and 92 can be ignored whenever the signal from detector 110 indicates that the size of the particles lies outside a range that particle detector 60 is optimized to detect (for example, based on the size of biological particles likely to correspond to pollen or paper particles, as opposed to biological toxins) and can be ignored. Furthermore, where laser diode 94 and its optical path are disposed in a plane above the optical paths for the LEDs and bio-fluorescence detectors, the signal from detector 110 can be used to trigger activation of the LED light sources according to a predefined pattern. In this exemplary embodiment, the LEDs are not always energized while the detector is being used. While LEDs are generally relatively long-lived solid-state devices, the UV-LEDs that can be employed in detector 60 are not nearly as long-lived as more conventional visible light LEDs. Thus, energizing the LEDs only when detector 110 indicates that aerosol particles are passing through the sample volume increases the operational lifespan of the LEDs and also reduces the overall power consumption of the detector. Furthermore, in some sampling paradigms, it will be preferable for each LED to be separately selectively energized, so that the autofluorescence of tryptophan and NADH are not simultaneously stimulated. This technique is particularly useful because the light emitted by the autofluorescence of tryptophan and the autofluorescence of NADH share some wavelengths, and separating the autofluorescence phenomenon in time enables the signals collected by detectors 86 and 92 to be more closely correlated with the autofluorescence of either tryptophan or NADH. In an exemplary, but not limiting embodiment, residence time of a particle in the sample volume is about 10−4 to about 10−31 seconds.
As noted above, detector 130 includes two primary components, which in an exemplary embodiment are implemented using monolithic structures. An illumination chamber component 146 can be implemented using two monolithic structures configured to join together in a facing relationship, generally as described above. While a single monolithic structure might be able to be employed, the use of two (or more) monolithic structures will likely result in considerable cost reductions. Illumination chamber component 146 includes multiple light sources, a sample volume, and a fluid path for the sheath flow and the gaseous fluid in which biological particles might be entrained. Generally as discussed above, relatively low cost non-imaging optical components are employed in illumination chamber component 146 to direct light from light sources toward the sample volume.
A detection flower component 148 includes a plurality of detectors, and a plurality of relatively low cost non-imaging optical components, which are employed to direct light from the sample volume in the illumination chamber component 146 toward specific detectors. In an exemplary embodiment, detection flower component 148 is implemented using two monolithic structures, which when joined together in a facing relationship define the detection flower component 148. The term detection flower refers to the floral like arrangement of a plurality of relatively similar petals or branches radiating from a central source.
Detector flower component 148 includes four branches (or petals). Preferably, each branch includes relatively low cost non-imaging optical components configured to direct light away from the sample volume and toward a detector. In a particularly preferred embodiment, detector flower component 148 is fabricated using two monolithic structures joined together in a facing relationship. Those of ordinary skill in the art will readily recognize that other combinations of monolithic structure can be employed (for example, each branch/petal can be formed as a single monolithic structure, or each branch/petal can be formed of two opposing monolithic structures).
A branch 156 includes a non-imaging optical component configured to direct light from illumination chamber component 146 toward a central core 164 of the detector flower. Optical elements (preferably dichroic filters or other types of beam splitters) in central core 164 direct light along branches 158, 160, and 162. As will be discussed in greater detail below, each branch 158, 160 and 162 includes a detector. Preferably, each branch 158, 160 and 162 also includes a CPC to help direct the light to the detector.
Three different light sources are employed in the illumination chamber component; consistent with the structure shown in
A second LED (which is hidden from view by monolithic structure 147, but which is disposed generally opposite LED 170) introduces light having a different wavelength into the sample volume. Two reflectors are similarly employed to direct a light beam 178 into the sample volume. Of those two reflectors, only the second (reflector 180) is visible in
Light designated λ1 corresponds to UV fluorescence, which is substantially reflected by filter 206 into CPC 204 to detector 193. Filter 196 is used to prevent stray light (i.e., scattered IR, scattered UV having a different wavelength than the wavelength detector 193 is configured to detect, and UV fluorescence having a different wavelength than the wavelength detector 193 is configured to detect). UV fluorescence corresponds to light emitted from a stimulated biological particle, and can be used to determine if biological materials are in the sample volume.
Light designated λ2 corresponds to UV fluorescence (having a different wavelength that the UV fluorescence directed into branch 162 toward detector 193), which substantially passes through filter 206, and moves through CPC 200 to detector 191. Filter 194 can be employed to prevent stray light (i.e., scattered IR, scattered UV having a different wavelength than the wavelength detector 191 is configured to detect, and UV fluorescence having a different wavelength than the wavelength detector 191 is configured to detect). Having multiple UV fluorescence signals enables additional information about specific biological particles to be determined.
Light designated λ3 corresponds to both IR scatter and UV scatter (UV light from the LEDs, as opposed to UV fluorescence emitted from biological particles). Filter 206 acts as a beam splitter for IR scatter and UV scatter, such that a portion of the light designated λ2 (i.e., about 50%) passes through filter 206 and moves toward filter 194, which has been selected to reflect IR scatter and UV scatter, while passing fluorescence of desired range of wavelengths (which correspond to wavelengths detector 191 is configured to detect). When the portion of light designated λ2 that is reflected by filter 194 reaches filter 206, a portion of that light is reflected by filter 206 toward CPC 202 and detector 192. When the light designated λ2 initially encounters filter 206, the portion that does not pass through filter 206 (and moves toward filter 194) is reflected by filter 206, and thus directed to filter 196, which has been selected to reflect IR scatter and UV scatter, while passing fluorescence of desired range of wavelengths (which correspond to wavelengths detector 193 is configured to detect). When the portion of light designated λ2 that is reflected by filter 196 reaches filter 206, a portion of that light passes through filter 206 and moves through CPC 202 and toward detector 192. In the detector flower, IR side scatter is used to detect when a particle (which could be biological) enters the sample volume. UV scatter information can be used to help determine sizing of particles in the sample volume.
Generally as described above, illumination chamber component 146a includes gaseous fluid inlet 134, sheath inlet 154, and vacuum pump 152, which cooperate to direct gaseous fluid 134a into sample volume 182. Also as generally described above, illumination chamber component 146a includes exit port 186, through which light of particular interest is directed toward detection components.
Illumination chamber component 146a includes LED 62 and CPC 66, which direct a light beam 62a through a beam combining reflector 71 toward a reflector 73, which directs light beam 62a toward sample volume 182. CPC 66 can be replaced by a parabolic reflector.
Illumination chamber component 146a includes LED 64 and CPC 68, which direct a light beam 64a toward beam combining reflector 71, which redirects light beam 64a toward reflector 73, which directs light beam 64a toward sample volume 182. CPC 68 can be replaced by a parabolic reflector.
Illumination chamber component 146a includes laser diode 184 (i.e., an IR laser source), which emits light beam 185 through reflector 73 toward sample volume 182. If the reflector is not transparent to IR wavelengths, an opening (not separately shown) can be formed into reflector 73 to enable IR light beam 185 to reach sample volume 182.
Illumination chamber component 146a also includes exit port 187, which serves several functions, including providing a beam dump for un-scattered IR and un-scattered UV (from LEDs 62 and 64). In some embodiments, exit port 187 also enables collection of forward particle (IR) scattering by a forward scatter mirror and detector (neither of which are separately shown). Note that in embodiments that include a forward scatter mirror and detector, the IR side-scatter is not required to be collected in the detection flower (i.e., the forward scatter is used to trigger the LEDs, not the side-scatter).
Note that a portion of reflector 73 intrudes into illumination chamber 151. Empirical testing of this configuration indicates that having reflector 73 protrude into the illumination chamber appears to cause unintentional scattering of both UV and IR light. The unintentionally scattered light exits the illumination chamber along with light of particular interest via exit port 186, and thus the unintentionally scattered light reaches the detectors and is responsible for undesirable levels of noise. Note that light scattered from particles in the sample volume is light of particular interest. However, light scattered by reflector 73 is not, and simply represents noise.
Generally as described above, illumination chamber component 210 includes a gaseous fluid inlet 134, sheath inlet 154 (not shown), and vacuum pump 152 (only a portion of which is shown), which cooperate to direct gaseous fluid 134a into sample volume 182. Also as generally described above, illumination chamber component 146a includes exit port 186, through which light of particular interest is directed toward detection components (such as a detection flower, or the detection elements of
Illumination chamber component 210 includes an LED 212 and a parabolic reflector 215, which directs a light beam 217 through a filter 220 toward a reflector 224, which directs light beam 217 toward sample volume 182. Preferably, reflector 215 is not a CPC.
Illumination chamber component 210 includes LED 214 and a parabolic reflector 216, which directs a light beam 219 toward filter 220, which redirects light beam 219 toward reflector 224, which directs light beam 219 toward sample volume 182. Preferably, reflector 216 is not a CPC.
Illumination chamber component 210 includes laser diode 184 (i.e., an IR laser source), which emits light beam 185 through reflector 224 toward sample volume 182. If the reflector is not transparent to IR wavelengths, an opening (not separately shown) can be formed into reflector 224 to enable IR light beam 185 to reach sample volume 182.
Illumination chamber component 210 also includes exit port 187, which serves several functions, including providing a beam dump for un-scattered IR and un-scattered UV (from LEDs 212 and 214). In this embodiments, exit port 187 also enables collection of forward particle (IR) scattering by a forward scatter mirror 228 and a detector (not separately shown).
Note that no portion of filter 224, nor any other element employed to direct light into the sample volume, intrudes into illumination chamber 151, which will reduce the amount of unintentionally scattered light reaching the UV scatter and fluorescence detectors, thereby reducing noise.
Additional noteworthy changes include employing a relatively long focal length mirror for reflector 224, which will minimize beam divergence of the light from the LEDs. A primary aperture screen 220 blocks stray light from the LEDs, and a secondary aperture screen 226 blocks scattered light.
Although the concepts disclosed herein have been described in connection with the preferred form of practicing them and modifications thereto, those of ordinary skill in the art will understand that many other modifications can be made thereto within the scope of the claims that follow. Accordingly, it is not intended that the scope of these concepts in any way be limited by the above description, but instead be determined entirely by reference to the claims that follow.
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/US07/67554 | 4/26/2007 | WO | 00 | 1/23/2009 |
Number | Date | Country | |
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60714007 | Sep 2005 | US |
Number | Date | Country | |
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Parent | 11380629 | Apr 2006 | US |
Child | 12298744 | US |