Patent Application
20240402477
The subject matter relates generally to optical detection systems for processing biological materials and more particularly to multiscale lens systems and methods for imaging well plates and including event-based detection.
Optical detection systems are used for processing biological materials in well plates. Examples of well plates include, but are not limited to, 6-well, 12-well, 24-well, 48-well, 96-well, 384-well, and 1536-well microplates. Currently, existing optical detection processes scan from well to well or from one position to another position and take images with a camera. Accordingly, existing optical detection systems can be complex because they require moving parts. Further, current optical detection processes are slow because the well plates are imaged and processed one well at a time. For example, if it takes about one second to image and process one well, it may take up to about 384 seconds (more than 6 minutes) to perform optical detection on a 384-well microplate. Therefore, new approaches are needed with respect to simplifying optical detection systems and/or speeding up the optical detection process for well plates.
Aspects of the disclosure provided herein comprise a system configured to image a surface, comprising: a first light source and a second light source configured to illuminate a first surface of a well, wherein the first light source is separated at a distance of less than about 500 nanometers from the second light source: an optical element configured to couple emitted light of the first light source and the second light source from a second surface of the well, wherein the second surface of the well is axially separated from the first surface of the first well along an optical axis parallel to an optical axis of the first light source and the second light source, and wherein the optical element comprises a plurality of lens elements; and a detector optically coupled to the optical element, wherein the detector is configured to detect the light emitted from the second surface of the well. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the first light source and the second light source comprise a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the well comprises a well of a multi-well plate. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first well of the multi-well plate, and a second detector of the plurality of detectors is configured to detect light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the first light source emits a first wavelength of light, and the second light source emits a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provided herein comprise a system configured to image a surface, comprising: a first light source and a second light source configured to illuminate a first surface of a well, wherein the first light source is positioned at a distance of less than about the diffraction limit of the second light source: an optical element configured to couple emitted light of the first light source and the second light source from a second surface of the well, wherein the second surface of the well is axially separated from the first surface of the first well along an optical axis parallel to an optical axis of the first light source and the second light source, and wherein the optical element comprises a plurality of lens elements; and a detector optically coupled to the optical element, wherein the detector is configured to detect the light emitted from the second surface of the well. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the first light source and the second light source comprise a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the well is a well of a multi-well plate. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first well of the multi-well plate, and a second detector of the plurality of detectors is configured to detect light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the first light source emits a first wavelength of light and the second light source emits a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provided herein comprise a system configured to image a surface, comprising: a first light source and a second light source configured to illuminate a first surface of a well: an optical element configured to couple emitted light of the first light source and the second light source from a second surface of the well, wherein the second surface of the well is axially separated from the first surface of the first well along an optical axis parallel to an optical axis of the first light source and the second light source, wherein the optical element comprises a first optical element and a second optical element separated by a center-to-center spacing of at least 100 nanometers; and a detector optically coupled to the optical element, wherein the detector is configured to detect the light emitted from the second surface of the well. In some embodiments, first optical element and the second optical element are lens elements. In some embodiments, the optical element comprises a plurality of lens elements. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the first light source and the second light source comprise a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the well is a well of a multi-well plate. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first well of the multi-well plate, and a second detector of the plurality of detectors is configured to detect light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the first light source emits a first wavelength of light and the second light source emits a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provided comprise a system configured to image a surface, comprising: a first light source and a second light source configured to illuminate a first surface of a well: an optical element configured to couple emitted light of the first light source and the second light source from a second surface of the well, wherein the second surface of the well is axially separated from the first surface of the first well along an optical axis parallel to an optical axis of the first light source and the second light source, wherein the optical element comprises a first optical element and a second optical element which are separated by a distance of less than about the diffraction limit of light emitted by the first light source or the second light source; and a detector optically coupled to the optical element, wherein the detector is configured to detect the light emitted from the second surface of the well. In some embodiments, the first optical element and the second optical element are lens elements. In some embodiments, the optical element comprises a plurality of lens elements. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the first light source and the second light source comprise a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the well is a well of a multi-well plate. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first well of the multi-well plate, and a second detector of the plurality of detectors is configured to detect light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the first light source emits a first wavelength of light and the second light source emits a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provided comprise a method, comprising: illuminating a first surface of a well with a first light source and a second light source, wherein the first light source is separated by at least about 500 nm from the second light source; and detecting light emitted from a second surface of the well through an optical element optically coupled to the second surface, wherein the second surface of the well is axially separated from the first surface of the well along an optical axis parallel to the first light source and the second light source. In some embodiments, the light emitted from the second surface is detected by a detector. In some embodiments, the detector comprises a plurality of detectors. In some embodiments, the detector comprises a two-dimensional sensor. In some embodiments, the detector comprises a charge coupled device sensor or a complementary metal oxide semiconductor sensor. In some embodiments, the well comprises a well of a multi-well plate. In some embodiments, the method further comprises translating the detector, first light source, and second light source along an axis of the multi-well plate and repeating (a) and (b). In some embodiments, the optical element comprises a first optical element and a second optical element. In some embodiments, the first optical element and the second optical element are lens elements. In some embodiments, the optical element comprises a plurality of lens elements. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the first light source and the second light source comprise a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first well of the multi-well plate, and a second detector of the plurality of detectors is configured to detect light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the first light source emits a first wavelength of light and the second light source emits a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provided comprise a method, comprising: illuminating a first surface of a well with a first light source and a second light source, wherein the first light source is separated by at least about the diffraction limit of the first light source or the second light source; and detecting light emitted from a second surface of the well through an optical element optically coupled to the second surface, wherein the second surface of the well is axially separated from the first surface of the well along an optical axis parallel to the first light source and the second light source. In some embodiments, the light emitted from the second surface is detected by a detector. In some embodiments, the detector comprises a plurality of detectors. In some embodiments, the detector comprises a two-dimensional sensor. In some embodiments, the detector comprises a charge coupled device sensor or a complementary metal oxide semiconductor sensor. In some embodiments, the well comprises a well of a multi-well plate. In some embodiments, the method further comprises translating the detector, first light source, and second light source along an axis of the multi-well plate and repeating (a) and (b). In some embodiments, the optical element comprises a first optical element and a second optical element. In some embodiments, the first optical element and the second optical element are lens elements. In some embodiments, the optical element comprises a plurality of lens elements. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the first light source and the second light source comprise a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first well of the multi-well plate, and a second detector of the plurality of detectors is configured to detect light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the first light source emits a first wavelength of light and the second light source emits a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provide comprise a method, comprising: illuminating a first surface of a well with a first light source and a second light source, wherein the; and detecting light emitted from a second surface of the well through an optical element optically coupled to the second surface, wherein the second surface of the well is axially separated from the first surface of the well along an optical axis parallel to the first light source and the second light source, and wherein the optical element comprises a first optical element and a second optical element separated by a distance of at least about 100 nanometers. In some embodiments, the light emitted from the second surface is detected by a detector. In some embodiments, the detector comprises a plurality of detectors. In some embodiments, the detector comprises a two-dimensional sensor. In some embodiments, the detector comprises a charge coupled device sensor or a complementary metal oxide semiconductor sensor. In some embodiments, the well comprises a well of a multi-well plate. In some embodiments, the method further comprises translating the detector, first light source, and second light source along an axis of the multi-well plate and repeating (a) and (b). In some embodiments, the first optical element and the second optical element are lens elements. In some embodiments, the optical element comprises a plurality of lens elements. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the first light source and the second light source comprise a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first well of the multi-well plate, and a second detector of the plurality of detectors is configured to detect light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the first light source emits a first wavelength of light and the second light source emits a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provided comprise a method, comprising: illuminating a surface of a well with a light source, wherein the surface of the well comprises at least a first analyte and a second analyte disposed adjacent to the surface, wherein the first analyte and the second analyte are separated by a distance of less than about the diffraction limit of light of a first emission of the first analyte or the diffraction limit of light of a second emission of the second analyte emitted when illuminated by the light source; and detecting at least a signal of the first emission and of the second emission from the surface of the well through an optical element optically coupled to the surface, wherein the optical element comprises at least a first optical element and a second optical element separated by a distance of less than about the diffraction limit of the light of the first emission or the light of the second emission. In some embodiments, the light of the first emission and the second emission are detected by a detector. In some embodiments, the detector comprises a plurality of detectors. In some embodiments, the detector comprises a two-dimensional sensor. In some embodiments, the detector comprises a charge coupled device sensor or a complementary metal oxide semiconductor sensor. In some embodiments, the well comprises a well of a multi-well plate. In some embodiments, the method further comprises translating the detector, light source, and optical element along an axis of the multi-well plate and repeating (a) and (b). In some embodiments, the first optical element and the second optical element are lens elements. In some embodiments, the optical element comprises a plurality of lens elements. In some embodiments, the optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the light source comprises a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect a signal of light from a first well of the multi-well plate, and a second detector of the plurality of detectors configured to detect a signal of light from a second well of the multi-well plate. In some embodiments, the first well and the second well are non-adjacent wells of the multi-well plate. In some embodiments, the light source emits a first wavelength of light and a second wavelength of light, wherein the first wavelength of light and the second wavelength of light differ.
Aspects of the disclosure provided comprise a system configured to image a surface, comprising: a light source configured to illuminate at least a first surface of a first well and a first surface of a second well of a multi-well plate, wherein the first well and the second well are nonadjacent wells: a first optical element configured to transmit emitted light of a second surface of the first well and a second surface of the second well to an imaging plane of a second optical element, wherein the first optical element comprises a plurality of lens elements, wherein the first surface of the first well is axially separated from the second surface of the first well along an optical axis of the light source, and wherein the first surface of the second well is axially separated from the second surface of the second well along the optical axis of the light source; and a detector optically coupled to the second optical element, wherein the detector is configured to detect the emitted light of the second surface of the first well and the second surface of the second well. In some embodiments, the plurality of lens elements of the first optical element further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the plurality of lens elements comprise at least a first lens element configured to transmit emitted light of the second surface of the first well to the imaging plane of the second optical element, and a second lens element configured to transmit emitted light of the second surface of the second well to the imaging plane of the second optical element. In some embodiments, the light source comprises a light emitting diode. In some embodiments, the light emitting diode comprises a plurality of light emitting diodes. In some embodiments, the light emitting diode comprises an array of light emitting diodes. In some embodiments, the detector comprises a plurality of detectors, wherein a first detector of the plurality of detectors is configured to detect emitted light from a first area of the well plate, wherein a second detector of the plurality of detectors is configured to detect emitted light from a second area of the well plate, and wherein the first area of the well plate and the second area of the well plate do not overlap. In some embodiments, the detector comprises a two-dimensional sensor. In some embodiments, the detector comprises a charge coupled device sensor or a complementary metal oxide semiconductor sensor. In some embodiments, the light source comprises a first light source configured to illuminate the first surface of the first well and a second light source configured to illuminate the second surface of the second well. In some embodiments, the first light source and the second light source emit wavelengths or wavelength bands of light that differ. In some embodiments, the second optical element comprises a plurality of optical elements. In some embodiments, the second optical element comprises an aspheric doublet, a positive focal length lens, a negative focal length lens, a plano-convex lens, a plano-concave lens, a convex lens, a bi-convex lens, a concave lens, a bi-concave lens, or any combination thereof.
Aspects of the disclosure provided comprise a method, comprising: providing a multi-well plate, wherein the multi-well plate comprises a plurality of wells: illuminating at least a first surface of a first well and a first surface of a second well of the plurality of wells, wherein the first well and the second well are not adjacent; and detecting light emitted from a second surface of the first well and a second surface of the second well through an optical element, wherein the first surface of the first well is axially separated from the second surface of the first well along an optical axis of the light emitted, and wherein the first surface of the second well is axially separated from the second surface of the second well along the optical axis of the light emitted. In some embodiments, the light emitted from the second surface of the first well and the second surface of the second well is detected by a detector. In some embodiments, the detector comprises a plurality of detectors. In some embodiments, the method further comprises translating the detector along an axis of the multi-well plate and repeating (b) and (c). In some embodiments, the detector comprises a two-dimensional sensor. In some embodiments, the detector comprises a charge coupled device sensor or a complementary metal oxide semiconductor sensor. In some embodiments, the optical element comprises at least a first optical element and a second optical element, wherein the first optical element is configured to detect light emitted from the second surface of the first well, and wherein the second optical element is configured to detect light emitted from the second surface of the second well. In some embodiments, the optical element comprises a plurality of lens elements. In some embodiments, the plurality of lens elements further comprises a polygonal optical element coupled to a surface of at least a lens of the plurality of lens elements. In some embodiments, the optical element comprises a micro-lens array. In some embodiments, the optical element comprises a plurality of gradient index of refraction (GRIN) lenses. In some embodiments, the first surface of the first well and the first surface of the second well are illuminated by a light source. In some embodiments, the light source comprises a light emitting diode. In some embodiments, the light source comprises a plurality of light emitting diodes. In some embodiments, the light source comprises an array of light emitting diodes. In some embodiments, the light source comprises a first light source configured to illuminate the first surface of the first well and a second light source configured to illuminate the first surface of the second well. In some embodiments, the first light source emits a first wavelength or first wavelength band, wherein the second light source emits a second wavelength or second wavelength band, and wherein the first wavelength or first wavelength band differs from the second wavelength or second wavelength band.
Having thus described the subject matter in general terms, reference will now be made to the accompanying drawings, which are not necessarily drawn to scale, and wherein:
The subject matter now will be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the subject matter are shown. Like numbers refer to like elements throughout. The subject matter may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein: rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Indeed, many modifications and other embodiments of the subject matter set forth herein will come to mind to one skilled in the art to which the subject matter pertains having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the subject matter is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims.
In some embodiments, the subject matter provides multiscale lens systems and methods for imaging well plates and including event-based detection. Further, a fluidics system is provided that may include at least one of the multiscale lens systems and a liquid handling system in relation to, for example, a multi-well microplate.
In some embodiments, the multiscale lens systems and methods provide an optical detection system including one or more read heads (e.g., cameras) that are stationary and accordingly an optical detection system is provided that includes no moving parts.
In some embodiments, the multiscale lens systems and methods provide an optical detection system including one or more read heads (e.g., cameras) that may be moved (or stepped) in two dimensions and accordingly an optical detection system is provided that may include certain moving parts, such as a camera system mounted on a linear translator (e.g., linear X-Y stage).
In some embodiments, the multiscale lens systems and methods provide a six read-head optical detection system to perform high resolution fluorescence imaging of all wells of a well plate, such as, but not limited to, all wells of a 384-well microplate.
In some embodiments, the multiscale lens systems and methods provide a six read-head optical detection system and wherein the combined fields-of-view (FOV) of six stationary read heads (e.g., cameras) may be used to image a well plate substantially in its entirety.
In some embodiments, the multiscale lens systems and methods provide a six read-head optical detection system for imaging an entire well plate using a single image capture operation at each of the six stationary read heads (e.g., cameras) and then processing the six images to analyze the biological materials present in each individual well of the well plate. In some embodiments, the biological materials may comprise one or more analytes, cells, nucleic acid molecules, proteins, peptides, beads, or any combination thereof.
In some embodiments, the multiscale lens systems and methods provide a step-and-shoot optical detection system to perform high resolution fluorescence imaging of all wells of a well plate, such as, but not limited to, all wells of a 384-well microplate.
In some embodiments, the multiscale lens systems and methods provide a 6-step step-and-shoot optical detection system and wherein a single read head (e.g., camera) may be stepped in X-Y to sequentially image a one-sixth portion of a well plate.
In some embodiments, the multiscale lens systems and methods provide a 24-step step-and-shoot optical detection system and wherein a single read head (e.g., camera) may be stepped in X-Y to sequentially image a one-twenty-fourth portion of a well plate.
In some embodiments, the multiscale lens systems and methods provide a step-and-shoot optical detection system and wherein a single read head (e.g., camera) may be stepped in X-Y to sequentially image portions of a well plate and wherein each image may be processed and analyzed with respect to determining the biological materials present in each individual well of the well plate.
In some embodiments, the multiscale lens systems and methods provide a multiscale lens array optical detection system to perform high resolution fluorescence imaging of all wells of a well plate, such as, but not limited to, all wells of a 384-well microplate.
In some embodiments, the multiscale lens systems and methods provide a multiscale microlens optical detection system including a microlens array, a controllable light source array, and a stationary read head (e.g., camera), all in relation to a well plate.
In some embodiments, the multiscale lens systems and methods provide a multiscale microlens optical detection system including an array of microlenses and a large-scale compound lens and wherein each of the microlenses and the large-scale compound lens work independently.
In some embodiments, the multiscale lens systems and methods provide a multiscale microlens optical detection system including an array of microlenses and a large-scale compound lens and thereby providing a way to have both a large numerical aperture (NA) and a large field-of-view (FOV).
In some embodiments, the multiscale lens systems and methods provide a multiscale gradient-index (GRIN) lens optical detection system including a GRIN lens array, a controllable light source array, and a stationary read head (e.g., camera), all in relation to a well plate.
In some embodiments, the multiscale lens systems and methods provide a multiscale GRIN lens optical detection system including an array of GRIN lenses and a large-scale compound lens and wherein each of the GRIN lenses and the large-scale compound lens work independently.
In some embodiments, the multiscale lens systems and methods provide a multiscale GRIN lens optical detection system including an array of GRIN lenses and a large-scale compound lens and thereby providing a way to have both a large numerical aperture (NA) and a large field-of-view (FOV).
In some embodiments, the multiscale lens systems and methods provide a fluidics system that may include a six read-head optical detection system or a step-and-shoot optical detection system or a multiscale microlens optical detection system or a multiscale GRIN lens optical detection system and a liquid handling system in relation to a well plate.
In some embodiments, the multiscale lens systems and methods provide a fluidics system that may include a six read-head optical detection system or a step-and-shoot optical detection system or a multiscale microlens optical detection system or a multiscale GRIN lens optical detection system and wherein the optical detection systems may further include event-based sensors.
In some embodiments, the multiscale lens systems and methods provide a fluidics system that may include a six read-head optical detection system or a step-and-shoot optical detection system or a multiscale microlens optical detection system or a multiscale GRIN lens optical detection system, a controller, and an image processing module or algorithm.
Further, a method of using a fluidics system including a multiple read-head optical detection system is provided.
Further, a method of using a fluidics system including a step-and-shoot optical detection system is provided.
Further, a method of using a fluidics system including a lens array optical detection system is provided.
Referring now to
Further, a liquid handling system 190 may be provided in relation to well plate 180. Liquid handling system 190 may be, for example, an automated liquid handling system. In this example, six read-head optical detection system 100 may be provided beneath well plate 180 and liquid handling system 190 may be provided above well plate 180. Well plate 180 may be any standard glass or plastic well plate used for processing biological materials and that is substantially transparent to light. Well plate 180 may be, for example, a standard 6-well, 12-well, 24-well, 48-well, 96-well, 384-well, or 1536-well microplate. In the example shown in
Six read-head optical detection system 100 may include six camera systems 110. For example, six read-head optical detection system 100 may include camera systems 110a, 110b, 110c, 110d, 110e, and 110f arranged in a 2×3 configuration. Each of the camera systems 110 may be capable of performing high resolution fluorescence imaging. For example, each of the camera systems 110 may include a large-scale compound lens, an image sensor, and a light source among other optical components. Generally, each of the camera systems 110 may include any arrangements of optical components, such as, but not limited to, optical lenses, optical filters, mirrors, prisms, polarizers, gratings, and the like. More details of example configurations of camera systems 110 are shown and described below with reference to
Each of the camera systems 110 of six read-head optical detection system 100 has a certain field-of-view (FOV). Together, the combined fields-of-view (FOVs) of the six camera systems 110 may capture the full area of well plate 180 with all its wells (e.g., 384 wells). For example,
In six read-head optical detection system 100, each of the camera systems 110 corresponds to one of the plate segments 184 of 384-well microplate 180. Further,
Referring now to
In operation, six read-head optical detection system 100 is held stationary with respect to, for example, 384-well microplate 180. Assays may be performed with one or more of the wells 182 of 384-well microplate 180. The assays (or chemistry) may be facilitated using liquid handling system 190. During a detection step of the workflow, each of the six camera systems 110 of six read-head optical detection system 100 may capture an image of its corresponding plate segment 184 of 384-well microplate 180. More specifically, at substantially the same time, each of the camera systems 110a, 110b, 110c, 110d, 110e, and 110f may take a snapshot of its corresponding plate segment 184 of 384-well microplate 180.
Accordingly, during the detection step, which is an image capture step, six digital images from six read-head optical detection system 100 are available for processing. Together the six images capture substantially the entirety of 384-well microplate 180 with its 384 wells 182. Next, each of the six digital images may be processed with respect to determining the contents of each individual well 182 of 384-well microplate 180. That is, certain pixels of each of the six digital images may be mapped to a specific well 182 of 384-well microplate 180. Then, each pixel group may be processed with respect to determining the contents of its corresponding well 182. An example of a fluidics system and method for processing digital images from six read-head optical detection system 100 is shown and described below in
Referring now to
In the configuration shown in
In this example, light source 120 is directed at dichroic mirror 114 that reflects the excitation light 130 into large-scale compound lens 112 and toward 384-well microplate 180, as shown in
Referring now to
In the configurations shown in
In the configurations of six read-head optical detection system 100 shown in
Further, in the configurations of six read-head optical detection system 100 shown in
Referring now to
This orientation of image sensors 116 may be needed because the size of each image sensor 116 may exceed the size of one plate segment 184 (e.g., about 34.50 mm×33.75 mm). In one example, in six read-head optical detection system 102, each image sensor 116 may be the Sony IMX661 Sensor available from Sony Corporation (Tokyo, Japan). The IMX661 Sensor is a large format 56.73 mm diagonal CMOS image sensor designed for industrial equipment. The IMX661 Sensor has a global shutter function and a high effective pixel count of 127.68 megapixels. Accordingly, the Sony IMX661 Sensor may provide a high-resolution image sensor that supports high resolution fluorescence imaging.
Further, in six read-head optical detection system 102, each camera system 110 uses its dichroic mirror 114 to direct emission light 132 toward its image sensor 116. Using camera system 110f as an example in
Referring still to
In six read-head optical detection system 102 shown in
Further, in six read-head optical detection system 102 the distance between 384-well microplate 180 and large-scale compound lens 112 of each camera system 110 may be about 84 mm. The overall length of large-scale compound lens 112 of each camera system 110 may be about 88 mm long. The lens diameter of large-scale compound lens 112 of each camera system 110 may be about 34 mm. Further,
Referring now to
The multiple read-head configurations are not limited to six camera systems 110 as shown in six read-head optical detection systems 100 and 102, other configurations of cameras are possible. For example, other multiple read-head optical detection systems may include twelve camera systems 110, twenty-four camera systems 110, up to one camera system 110 per well. However, these configurations may be driven by certain parameters, such as, but not limited to, cost, complexity, resolution/FOV of the image sensor, resolution that may be needed to do the required analysis, and so on.
Referring now to
Camera system 410 of 6-step step-and-shoot optical detection system 400 may be substantially the same as one camera system 110 of six read-head optical detection system 100 or 102. Camera system 410 may include, for example, a large-scale compound lens 412, a mirror 414, an image sensor 416 mounted on a PCB 418, and a light source 420 that has a focusing lens 422. In some cases, the mirror 414 may be a dichroic mirror, partially reflective mirror, or a beam splitter. Further, in this configuration, 6-step step-and-shoot optical detection system 400 may be arranged beneath and/or above a 384-well microplate 180. Further, liquid handling system 190 (not shown) may be arranged above and/or below the 384-well microplate 180. Further, light source 420 of camera system 410 may be arranged below or above well plate.
In 6-step step-and-shoot optical detection system 400 shown in
Further, a main feature of 6-step step-and-shoot optical detection system 400 may include fast six-shot imaging. For example, 6-step step-and-shoot optical detection system 400 may use six image capture events and then process the resulting six images to analyze, for example, all 384 wells 182 of the 384-well microplate 180. A process that may take about 10 seconds or less than about 10 seconds. By contrast, existing optical detection processes are slow because the well plates are imaged and processed one well at a time. For example, if it takes about one second to image and process one well, it may take up to about 384 seconds (more than 6 minutes) to perform optical detection on a 384-well microplate.
In 6-step step-and-shoot optical detection system 400, it may take six steps of camera system 410 to capture the entirety of a well plate. That is, the FOV (e.g., a FOV 486) of camera system 410 may be sized to capture a plate segment 484 of a well plate, such as 384-well microplate 180 as shown in
Referring now to
In other examples, for a 6-well microplate each plate segment 484 and step is a 1-well segment. For a 24-well microplate each plate segment 484 and step may be a 2×2-well segment. For a 96-well microplate each plate segment 484 and step may be a 4×4-well segment. For a 1536-well microplate each plate segment 484 and step may be a 16×16-well segment.
For example, during the detection step, six digital images from 6-step step-and-shoot optical detection system 400 may be generated and subsequently processed. Together the six images may capture substantially the entirety of 384-well microplate 180 with its 384 wells 182. Next, each of the six digital images may be processed with respect to determining the contents of each individual well 182 of 384-well microplate 180. That is, certain pixels of each of the six digital images may be mapped to a specific well 182 of 384-well microplate 180. Then, each pixel group may be processed with respect to determining the contents of its corresponding well 182. An example of a fluidics system including 6-step step-and-shoot optical detection system 400 is shown and described below in
Further, in 6-step step-and-shoot optical detection system 400, image capture operations may occur, for example, at a certain plate segment 484 of 384-well microplate 180 while at substantially the same time certain fluidics operations (using liquid handling system 190) may occur at different plate segments 484 of 384-well microplate 180 that are not being imaged. These simultaneous actions in 6-step step-and-shoot optical detection system 400 may image an entirety of a 384-well microplate and/or greater than a 384-well microplate in a shorter period of time (e.g., about or less than about 10 seconds) when compared with existing well plate optical detection processes that require several minutes of imaging time.
The 24-step step-and-shoot optical detection system 402, may take twenty-four steps of camera system 410 to capture the entirety of a well plate 180. That is, the FOV (e.g., a FOV 486) of camera system 410 may be sized to capture a plate segment 484 of a well plate, such as 384-well microplate 180 as shown in
In 24-step step-and-shoot optical detection system 402 shown in
Further, a main feature of 24-step step-and-shoot optical detection system 402 may include fast 24-shot imaging. For example, the 24-step step-and-shoot optical detection system 402 may use twenty-four image capture events to produce twenty-four images to analyze, for example, all 384 wells 182 of the 384-well microplate 180. A process that may take about 30 seconds. By contrast, existing optical detection processes are slow because the well plates are imaged and processed one well at a time. For example, if it takes about one second to image and process one well, it may take up to about 384 seconds (more than 6 minutes) to perform optical detection on a 384-well microplate.
In other examples, for a 24-well microplate each plate segment 484 and step may be a 1-well segment. For a 96-well microplate each plate segment 484 and step may be a 2×2-well segment. For a 1536-well microplate each plate segment 484 and step may be an 8×8-well segment.
For example, during the detection step, twenty-four digital images from 24-step step-and-shoot optical detection system 402 may be available for processing. Together the twenty-four images may capture substantially the entirety of 384-well microplate 180 with its 384 wells 182. Next, each of the twenty-four digital images may be processed with respect to determining the contents of each individual well 182 of 384-well microplate 180. That is, certain pixels of each of the twenty-four digital images may be mapped to a specific well 182 of 384-well microplate 180. Then, each pixel group may be processed with respect to determining the contents of its corresponding well 182. An example of a fluidics system including 24-step step-and-shoot optical detection system 402 is shown and described below in
Further, in 24-step step-and-shoot optical detection system 402, image capture operations may occur, for example, at a certain plate segment 484 of 384-well microplate 180 while at substantially the same time certain fluidics operations (using liquid handling system 190) may occur at different plate segments 484 of 384-well microplate 180 that are not being imaged. These simultaneous actions in 24-step step-and-shoot optical detection system 402 may save time in a manner similarly described elsewhere herein with respect to existing well plate optical detection processes.
In some embodiments, the optical system as described elsewhere herein, may comprise a high resolution (e.g., super resolution imaging at or below about the diffraction limit of light emitted from a biological material) step and shoot optical system (404, 406), as shown in
In some cases, the multiscale micro lens high resolution step and shoot optical detection system 406, as shown in
In some cases, the multiscale micro lens high resolution step and shoot optical detection system 406 (
In some cases, the multiscale GRIN lens high resolution step and shoot optical detection system 404 (
A multiscale optical system is a system having two or more lenses that work independently. Further, multiscale optical systems may provide a way to have both a large numerical aperture (NA) and a large field-of-view (FOV). Examples of multiscale optical detection systems are described below with reference to
Referring now to
Multiscale microlens optical detection system 500 may include, for example, a camera system 510, a microlens array 520, and a light source array 524. Further, camera system 510, microlens array 520, and light source array 524 may be arranged with respect to a well plate, such as 384-well microplate 180. In one example, camera system 510 and microlens array 520 may be arranged as shown beneath 384-well microplate 180. While light source array 524 may be arranged as shown above the 384-well microplate 180. Further, liquid handling system 190, described elsewhere herein, may be arranged above 384-well microplate 180.
Microlens array 520 may include one microlens 522 per well of the well plate. For example, if the well plate is the 384-well microplate 180 as shown in
Similarly, light source array 524 may include one LED 526 per well of the well plate. For example, if the well plate is the 384-well microplate 180 as shown in
Light source array 524 provides an addressable LED array that may be used for virtual scanning. For example, each well 182 of 384-well microplate 180 may be addressable with a small LED 526 from light source array 524. In other embodiments, more complicated illumination systems may be possible. In one example, epifluorescence illumination may be integrated into the compound optics of camera system 510 (e.g., epifluorescence illumination compound microscope technology). In another example, the wells may be addressed by imaging a pattern onto the well plate surface using, for example, a spatial light modulator (SLM).
Camera system 510 may include, for example, a large-scale compound lens 512 and an image sensor 516 mounted on a PCB 518 (see
Further, an intermediate image plane 530 is present within multiscale microlens optical detection system 500. For example, intermediate image plane 530 is between microlens array 520 and camera system 510.
From the well-side of intermediate image plane 530, each microlens 522 is used to focus an image of one well 182 onto intermediate image plane 530. For example, if a piece of paper were at the intermediate image plane 530, in image may be projected on the paper. Next, from the camera system-side of intermediate image plane 530, large-scale compound lens 512 (e.g., a larger optic) is focused on the intermediate image plane 530. Accordingly, large-scale compound lens 512 of camera system 510 may be used to image the image at intermediate image plane 530.
An advantage is this multiscale configuration is that each of the microlenses 522 can be high NA and collect a lot of light. Then, use microlenses 522 to magnify the images of each well 182 to the intermediate image plane 530. That is, on the well-side of the microlenses 522 the NA is high, while on the camera system-side the NA is lower. This allows large-scale compound lens 512 to have a low NA. Yet, at the same time, the system as a whole is still able to capture a large amount of light. That is, the presence of microlens array 520 allows more light to be captured than using the large-scale compound lens 512 alone. More details of multiscale microlens optical detection system 500 are shown and described below in
Referring now to
Here, using the addressable light source array 524 that is provided in relation to, for example, 384-well microplate 180, each well 182 may be illuminated with its own LED 526. Accordingly, the detection process uses selective well illumination via the addressable light source array 524 so that there is substantially no sub-image overlap. This allows, for example, multiple wells 182 of 384-well microplate 180 to be imaged and/or processed simultaneously. Therefore, the amount of time that may be needed to perform the optical detection process may be minimized.
In multiscale microlens optical detection system 500, light source array 524 may be used to, for example, selectively illuminate every nth well 182 of 384-well microplate 180. For example,
Then, the large-scale compound lens 512 of camera system 510 relays an intermediate image plane image 534 of multiple sub-images 532 at intermediate image plane 530 onto image sensor 516. Because of the magnification of microlenses 522, a lower pixel count sensor may be utilized as compared to, for example, the image sensor 116 of six read-head optical detection systems 100, 102.
Next,
Next,
Further, in multiscale microlens optical detection system 500, image capture operations may occur, for example, at a certain portion of 384-well microplate 180 while at substantially the same time certain fluidics operations (using liquid handling system 190) may occur at different portions of 384-well microplate 180 that are not being imaged (
Referring now to
In this example, microprism array 528 or other micro-optics may be used to reroute the well sub-images 532 from the periphery of the well plate closer to center of intermediate image plane 530. Further, in microprism array 528 the shapes of the microprisms 529 change corresponding to their positions with respect to the well plate. Benefits of this configuration may include (1) it provides the ability to shrink the x-y of intermediate image plane 530, and (2) it reduces the FOV requirements on the large-scale compound lens 512 of camera system 510.
Again, the detection process uses selective well illumination so that there is substantially no overlap of sub-images 532. Again, this allows, for example, multiple wells 182 of 384-well microplate 180 to be processed simultaneously, and therefore shortens the optical detection process.
In of multiscale microlens optical detection system 500 shown in
Generally, multiscale microlens optical detection system 500 shown in
Referring now to
GRIN lens array 554 may include one GRIN lens 556 per well of the well plate. For example, if the well plate is the 384-well microplate 180 as shown in
Referring now to
In multiscale GRIN lens optical detection system 550 shown in
Further, in other embodiments, multiscale GRIN lens optical detection system 550 shown in
Further, in multiscale GRIN lens optical detection system 550, image capture operations may occur, for example, at a certain portion of 384-well microplate 180 while at substantially the same time certain fluidics operations (using liquid handling system 190) may occur at different portions of 384-well microplate 180 that are not being imaged (
Referring now to
In one example, GRIN lens 600 may be the G1P10 series GRIN lens. The G1P10 series GRIN lens is a GRIN lens that is designed for imaging applications. Certain specifications of G1P10 series GRIN lens may include Ø1.0 mm, L=3.758 mm, WD=0.20 mm (Water and Dry), NA=0.5, and uncoated.
Other features of GRIN lens 600 may include (1) GRIN lens: flat glass rod, (2) focusing from gradient in refractive index, (3) small size (up to ˜2 mm), (4) high NA: up to 0.55, and (5) NA decreases with larger FOV.
Referring now to
In some embodiments, the multiscale optical detection system, described elsewhere herein, may comprise a high resolution microlens optical detection system 560 (
In some embodiments, the high resolution microlens optical detection system 560 (
In some embodiments, the high resolution GRIN lens optical detection system 562 (
Referring now to
Generally, fluidics system 700 may be any optical measurement system that can be used to accurately determine the presence or absence of a defined analyte and/or target component in different materials and to sensitively quantify the amount of analyte and/or target components present in a sample. In this example, fluidics system 700 may include well plate 180, liquid handling system 190, and any one or more of the multiscale optical detection systems 100, 102, 400, 402, 500, 550. Fluidics system 700 may also include a controller 710. Controller 710 may further include an image processing module 712 and digital images 714 generated by any of the multiscale optical detection systems 100, 102, 400, 402, 500, 550.
In fluidics system 700, liquid handling system 190 may be provided in relation to well plate 180. Again, liquid handling system 190 may be, for example, an automated liquid handling system. In this example, any one or more of the multiscale optical detection systems 100, 102, 400, 402, 500, 550 may be provided beneath well plate 180, while liquid handling system 190 may be provided above well plate 180. Again, well plate 180 may be any standard glass or plastic well plate used for processing biological materials and that is substantially transparent to light. Well plate 180 may be, for example, a standard 6-well, 12-well, 24-well, 48-well, 96-well, 384-well, or 1536-well microplate. The 384-well microplate 180 shown in
Controller 710 may be electrically connected to liquid handling system 190 and the multiscale optical detection systems 100, 102, 400, 402, 500, 550. Controller 710 may, for example, be a general-purpose computer, special purpose computer, personal computer, microprocessor, or other programmable data processing apparatus. Controller 710 may provide processing capabilities, such as storing, interpreting, and/or executing software instructions, as well as controlling the overall operations of fluidics system 700. The software instructions may include machine readable code stored in non-transitory memory that is accessible by controller 710 for the execution of the instructions. Controller 710 may be configured and programmed to control data and/or power aspects of fluidics system 700. Further, data storage (not shown) may be built into or provided separate from controller 710.
Multiscale optical detection systems 100, 102, 400, 402, 500, 550 may be used to obtain light intensity readings. In the presently disclosed optical detection configurations and/or systems shown in
In fluidics system 700, any one of the multiscale optical detection systems 100, 102, 400, 402, 500, 550 may be used to perform optical detection operations with respect to well plate 180. More details of examples of optical detection methods are shown and described below in
Referring now to
Further, by way of example, the steps of method 800 are described below with respect to using six read-head optical detection system 100 of
At a step 810, a fluidics system is provided that includes a liquid handling system and a six read-head optical detection system in relation to a well plate. In one example, fluidics system 700 of
At a step 815, the liquid handling system is used to facilitate fluidic assays that occur in the wells of the well plate. For example, and referring to
At a step 820, one image capture operation of the six read-head optical detection system may be used to perform high resolution fluorescence imaging of all the wells of the well plate. For example, one image capture operation of six read-head optical detection system 100 of
More specifically, at substantially the same time, an image capture event occurs at each of the six read-heads (e.g., at each of camera systems 110a, 110b, 110c, 110d, 110e, and 110f) of six read-head optical detection system 100. The image capture events include activating the light sources 120. Then, and referring now fluidics system 700 of
At a step 825, the digital images of the six read-head optical detection system are processed and used for analysis of biological materials within each well of the well plate. For example, six digital images 714 from camera systems 110a, 110b, 110c, 110d, 110e, and 110f, respectively, of six read-head optical detection system 100 may be processed and used for analysis of biological materials within, for example, each well 182 of 384-well microplate 180. For example, certain pixels of each of the six digital images 714 may be mapped to a specific well 182 of 384-well microplate 180. Then, each pixel group may be processed with respect to determining the contents of its corresponding well 182. In one example, controller 710 and/or image processing module 712 of fluidics system 700 may be used to perform the image processing operations.
At a step 830, the results of the biological materials analysis for each well of well plate is indicated. For example, the results of the biological materials analysis for each well 182 of 384-well microplate 180 may be indicated or otherwise reported to any interested party or entity. In one example, controller 710 and/or image processing module 712 of fluidics system 700 may be used to indicate the results.
Referring now to
Further, by way of example, the steps of method 900 are described below with respect to using the 6-step step-and-shoot optical detection system 400 shown in
At a step 910, a fluidics system is provided that includes a liquid handling system and a step-and-shoot optical detection system in relation to a well plate. In one example, fluidics system 700 of
At a step 915 and running concurrently to step 920, one image capture operation of the step-and-shoot optical detection system may be used to perform high resolution fluorescence imaging of a first portion of the well plate. For example, one image capture operation of the 6-step step-and-shoot optical detection system 400 shown in
More specifically, linear translator 424 may be used to position camera system 410 at, for example, one of the six plate segments 484 of 384-well microplate 180 (see
Further, image capture operations may occur at a certain plate segment 484 of 384-well microplate 180 while at substantially the same time certain fluidics operations (step 920) may occur at a different plate segment 484 of 384-well microplate 180 (see
At a step 920 and running concurrently to step 915, the liquid handling system is used to facilitate fluidic assays that occur in certain other wells of the well plate. For example, liquid handling system 190 (e.g., an automated liquid handling system) may be used to facilitate fluidic assays (e.g., certain chemistry) that occur, for example, in certain plate segments 484 of 384-well microplate 180 other than those being imaged (see
Further, fluidics operations may occur at a certain plate segment 484 of 384-well microplate 180 while at substantially the same time certain image capture operations (step 915) may occur at a different plate segment 484 of 384-well microplate 180 (see
At a decision step 925, it is determined whether imaging is completed at all portions of the well plate. For example, it may be determined whether imaging is completed as all plate segments 484 of 384-well microplate 180 (see
At a step 930, the read-head is stepped to the next portion of the well plate. For example, and referring to
At a step 935 and running concurrently to step 940, one image capture operation of the step-and-shoot optical detection system may be used to perform high resolution fluorescence imaging of a next portion of the well plate. For example, one image capture operation of the 6-step step-and-shoot optical detection system 400 shown in
Further, image capture operations may occur at the selected plate segment 484 of 384-well microplate 180 while at substantially the same time certain fluidics operations (step 940) may occur at a different plate segment 484 of 384-well microplate 180 (see
At a step 940 and running concurrently to step 935, the liquid handling system is used to facilitate fluidic assays that occur in certain other portions of the well plate. For example, liquid handling system 190 (e.g., an automated liquid handling system) may be used to facilitate fluidic assays (e.g., certain chemistry) that occur, for example, in certain plate segments 484 of 384-well microplate 180 other than those being imaged (see
At a step 945, the digital images of the step-and-shoot optical detection system are processed and used for analysis of biological materials within each well of the well plate. For example, the six digital images 714 from the six steps of camera system 410 of 6-step step-and-shoot optical detection system 400 may be processed and used for analysis of biological materials within, for example, each well 182 of 384-well microplate 180. For example, certain pixels of each of the six digital images 714 may be mapped to a specific well 182 of 384-well microplate 180. Then, each pixel group may be processed with respect to determining the contents of its corresponding well 182. In one example, controller 710 and/or image processing module 712 of fluidics system 700 may be used to perform the image processing operations. Method 900 may proceed to step 950.
At a step 950, the results of the biological materials analysis for each well of well plate is indicated. For example, the results of the biological materials analysis for each well 182 of 384-well microplate 180 may be indicated or otherwise reported to any interested party or entity. In one example, controller 710 and/or image processing module 712 of fluidics system 700 may be used to indicate the results. Method 900 ends.
Referring now to
Further, by way of example, the steps of method 1000 are described below with respect to using the multiscale microlens optical detection system 500 shown in
At a step 1010, a fluidics system is provided that includes a liquid handling system and a multiscale lens array optical detection system in relation to a well plate. In one example, fluidics system 700 of
At a step 1015, a first portion of the well plate is illuminated and as a result images of individual wells are projected onto an intermediate image plane of the lens array optical detection system. For example, and referring now to
At a step 1020 and running concurrently to step 1025, one image capture operation of the intermediate image plane of the multiscale lens array optical detection system may be used to perform high resolution fluorescence imaging of a first portion of the well plate. For example, and referring now to
Further, in this step, image capture operations may occur at a certain portion of 384-well microplate 180 while at substantially the same time certain fluidics operations (step 1025) may occur at different portions of 384-well microplate 180 (
At a step 1025 and running concurrently to step 1020, the liquid handling system is used to facilitate fluidic assays that occur in certain other portions of the well plate. For example, liquid handling system 190 (e.g., an automated liquid handling system) may be used to facilitate fluidic assays (e.g., certain chemistry) that occur, for example, in certain portions of 384-well microplate 180 other than those being imaged (see
At a decision step 1030, it is determined whether imaging is completed at all portions of the well plate. For example, it may be determined whether imaging is completed as all portions of 384-well microplate 180 (see
At a step 1035, a next portion of the well plate is illuminated and as a result images of individual wells are projected onto an intermediate image plane of the lens array optical detection system. For example, and referring now to
At a step 1040 and running concurrently to step 1045, one image capture operation of the intermediate image plane of the multiscale lens array optical detection system may be used to perform high resolution fluorescence imaging of a next portion of the well plate. For example, and referring now to
Further, in this step, image capture operations may occur at a certain portion of 384-well microplate 180 while at substantially the same time certain fluidics operations (step 1025) may occur at different portions of 384-well microplate 180 (
At a step 1045 and running concurrently to step 1040, the liquid handling system is used to facilitate fluidic assays that occur in certain other portions of the well plate. For example, liquid handling system 190 (e.g., an automated liquid handling system) may be used to facilitate fluidic assays (e.g., certain chemistry) that occur, for example, in certain portions of 384-well microplate 180 other than those being imaged (see
At a step 1050, the digital images of the multiscale lens array optical detection system are processed and used for analysis of biological materials within each well of the well plate. For example, the multiple intermediate image plane images 534, which are images of the multiple sets of every third well 182 of 384-well microplate 180 being illuminated, may be processed, and used for analysis of biological materials within, for example, each well 182 of 384-well microplate 180. For example, certain pixels of each of the intermediate image plane images 534 may be mapped to a specific well 182 of 384-well microplate 180. Then, each pixel group may be processed with respect to determining the contents of its corresponding well 182. In one example, controller 710 and/or image processing module 712 of fluidics system 700 may be used to perform the image processing operations. Method 1000 may proceed to step 1055.
At a step 1055, the results of the biological materials analysis for each well of well plate is indicated. For example, the results of the biological materials analysis for each well 182 of 384-well microplate 180 may be indicated or otherwise reported to any interested party or entity. In one example, controller 710 and/or image processing module 712 of fluidics system 700 may be used to indicate the results. Method 1000 ends.
In another example, any of the multiscale optical detection systems 100, 102, 400, 402, 500, 550 of fluidics system 700 shown in
Because event-based fluidics system 700 may be used for processing biological materials in well plates, event-based sensor 720 may be useful for event-based optical detection with respect to processing and/or assaying biological materials.
For example, image sensors 116 of six read-head optical detection systems 100 and 102 may be event-based sensors 720. Image sensors 416 of the 6-step step-and-shoot optical detection system 400 and/or of the 24-step step-and-shoot optical detection system 402 may be event-based sensors 720. Image sensors 516 of multiscale microlens optical detection system 500 and/or multiscale GRIN lens optical detection system 550 may be event-based sensors 720.
Any assay that uses light in a detection process may benefit from event-based vision. For example, event-based optical detection may be used in a fluorescence-based assay where incorporation of a fluorescently labeled molecule or “tag” is monitored. In another example, event-based optical detection may be used in an assay where an enzymatic reaction is used to generate light in response to a detection event (e.g., a fluorescent assay or a luminescent assay). In yet another example, event-based optical detection may be used in a colorimetric assay where the change of color can be registered as a change in intensity per pixel.
In one example, event-based optical detection may be used in a nucleic acid assay for analysis of one or more target nucleic acid sequences in a sample. Further, event-based optical detection may be extended to sequencing-by-synthesis (SBS) sequencing where incorporation of a fluorescently labeled nucleotide or enzymatic generation of light (e.g., pyrosequencing) are used.
In another example, event-based optical detection may be used in an immunodetection assay for analysis of one or more target analytes in a sample. For example, event-based optical detection may be extended to immunodetection of one or more target proteins in a sample. An immunodetection assay may, for example, be a colorimetric assay or non-colorimetric assay (e.g., a fluorescent assay or a luminescent assay).
In yet another example, event-based optical detection may be used in a multiomic assay that combines, for example, the detection of nucleic acid targets and protein targets in a single assay.
Event-based optical detection may be used for fluorescence lifetime imaging. For example, an analyte may be excited with a very short pulse of light (e.g., on the order of about 10{circumflex over ( )}-15 s to 10{circumflex over ( )}-9 s) and the pixels of the camera may detect counts in time bins after the pulse. An advantage of this approach is that fluorescence lifetime can be used to distinguish between analytes and because the signal is generated from pixels independently, they can be distinguished in real time thus enabling a real time process.
Accordingly, in event-based fluidics system 700 for processing and/or assaying biological materials, examples of “events” that may be detected using event-based sensor 720 may include, but are not limited to, the following.
Further, event-based optical detection using, for example, event-based sensor 720 may be useful in other ways with respect to systems for processing and/or assaying biological materials, such as, but not limited to, the following.
Accordingly, in some embodiments, the disclosure provides an event-based optical detection system (e.g., event-based fluidics system 700) that uses an event-based sensor (e.g., event-based sensor 720) in a system for processing and/or assaying biological materials.
In summary and referring now again to
Following long-standing patent law convention, the terms “a,” “an,” and “the” refer to “one or more” when used in this application, including the claims. Thus, for example, reference to “a subject” includes a plurality of subjects, unless the context clearly is to the contrary (e.g., a plurality of subjects), and so forth.
The terms “comprise,” “comprises,” “comprising,” “include,” “includes,” and “including,” are intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that may be substituted or added to the listed items.
Terms like “preferably,” “commonly,” and “typically” are not utilized herein to limit the scope of the claimed embodiments or to imply that certain features are critical or essential to the structure or function of the claimed embodiments. These terms are intended to highlight alternative or additional features that may or may not be utilized in a particular embodiment of the disclosure.
The term “substantially” is utilized herein to represent the inherent degree of uncertainty that may be attributed to any quantitative comparison, value, measurement, or other representation and to represent the degree by which a quantitative representation may vary from a stated reference without resulting in a change in the basic function of the subject matter at issue.
Various modifications and variations of the disclosed methods, compositions and uses of the disclosure will be apparent to the skilled person without departing from the scope and spirit of the disclosure. Although the subject matter has been disclosed in connection with specific preferred aspects or embodiments, it should be understood that the subject matter as claimed should not be unduly limited to such specific aspects or embodiments.
The subject matter may be implemented using hardware, software, or a combination thereof and may be implemented in one or more computer systems or other processing systems. In one aspect, the subject matter is directed toward one or more computer systems capable of carrying out the functionality described herein.
For the purposes of this specification and appended claims, unless otherwise indicated, all numbers expressing amounts, sizes, dimensions, proportions, shapes, formulations, parameters, percentages, quantities, characteristics, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about” even though the term “about” may not expressly appear with the value, amount, or range. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are not and need not be exact, but may be approximate and/or larger or smaller as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other existing factors depending on the desired properties sought to be obtained by the subject matter. For example, the term “about,” when referring to a value can be meant to encompass variations of, in some embodiments±100%, in some embodiments±50%, in some embodiments±20%, in some embodiments±10%, in some embodiments±5%, in some embodiments±1%, in some embodiments±0.5%, and in some embodiments±0.1% from the specified amount, as such variations are appropriate to perform the disclosed methods or employ the disclosed compositions.
Further, the term “about” when used in connection with one or more numbers or numerical ranges, should be understood to refer to all such numbers, including all numbers in a range and modifies that range by extending the boundaries above and below the numerical values set forth. The recitation of numerical ranges by endpoints includes all numbers, e.g., whole integers, including fractions thereof, subsumed within that range (for example, the recitation of 1 to 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, and the like) and any range within that range.
Although the foregoing subject matter has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be understood by those skilled in the art that certain changes and modifications can be practiced within the scope of the appended claims.
This application is a continuation application of International Application No. PCT/US2023/062536, filed Feb. 14, 2023, which claims the benefit of U.S. Provisional Application No. 63/310,514, filed on Feb. 15, 2022, and U.S. Provisional Application No. 63/480,574, filed on Jan. 19, 2023, each of which is herein incorporated by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| 63310514 | Feb 2022 | US | |
| 63480574 | Jan 2023 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/US2023/062536 | Feb 2023 | WO |
| Child | 18800619 | US |
