Clustered wavelength-division optical detection system and method of using the same

Abstract

This reduces the amount of light loss resulting from reflections by optical components when generating separate spectral ranges for light detection. [Solution] For example, a system for detecting light in a flow stream is described. A photodetection system according to a particular embodiment includes a wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, and first, second, and third photodetection modules configured to receive light in each of the first, second, and third predetermined spectral ranges, the photodetection modules having a plurality of photodetectors and optical components that transmit light having a predetermined subspectral range to the photodetectors. Systems and methods for measuring light emitted by a sample (e.g., in a flow stream), as well as kits having three or more wavelength separators, a plurality of photodetectors, and optical components are also provided.

Description

[Background technology]

[0001] Photodetection is often used to characterize the components of a sample (e.g., a biological sample) when the sample is used to diagnose a disease or medical condition. When a sample is irradiated, light can be scattered by the sample, transmitted through the sample, and emitted by the sample (e.g., by fluorescence). Differences in the components of the sample, such as morphology, absorptivity, and the presence of fluorescent labels, can cause differences in the light scattered, transmitted, or emitted by the sample. These differences can be used to characterize and identify the presence of components in the sample. To quantify these differences, light is collected and directed to the surface of a detector. The amount of light reaching the detector can affect the overall quality of the optical signal output by the detector. The amount of light reaching the detector can be increased by increasing the surface area of ​​the detector or by increasing the collection of light from the sample.

[0002] One technique that utilizes photodetection to characterize components in a sample is flow cytometry. The data generated from the detected light can be used to record the distribution of components and classify the desired material. A flow cytometer typically includes a sample reservoir for receiving fluid samples, such as blood samples, and a sheath reservoir for containing the sheath fluid.

[0003] A flow cytometer transports particles (including cells) in a fluid sample as a cell stream into a flow cell, while also directing a sheath fluid into the flow cell. Within the flow cell, a liquid sheath forms around the cell stream, giving it a substantially uniform velocity. The flow cell hydrodynamically focuses the cells in the stream so that they pass through the center of the light source within the flow cell. Light from the light source can be detected as scattered light or by transmission spectroscopy, or it can be absorbed by one or more components in the sample and re-emitted as emission. [Overview of the Initiative]

[0004] Aspects of the present disclosure include a clustered wavelength-division photodetector system having three or more wavelength separators that allow light having a predetermined spectral range to pass through. The present disclosure provides a photodetector system that needs to separate detected light into spectral ranges and reduce light reflection in order to generate multiple subspectral ranges to be detected by a photodetector in the system of interest. The inventors have found that reflection to generate light of distinct spectral ranges results in increased optical loss and, in certain examples, causes insufficient detector signal quality (e.g., a low signal-to-noise ratio). The present disclosure reduces the amount of optical loss resulting from reflection by optical components when generating distinct spectral ranges for photodetector. According to certain embodiments, as will be described in more detail below, the present disclosure includes the ability to generate light of 20 or more different spectral ranges while exhibiting optical loss of 20% or less, e.g., 19% or less, e.g., 18% or less, e.g., 17% or less, e.g., 16% or less, e.g., 15% or less, and the ability to generate light of 20 or more different spectral ranges while exhibiting optical loss of 10% or less. In some embodiments, the photodetector system is configured to generate two or more distinct spectral ranges, for example, three or more distinct spectral ranges of light, for each reflection by an optical component (e.g., a dichroic mirror). In a particular example, the photodetector system is configured to generate 30 different spectral ranges of light from 10 or fewer reflections by an optical component, for example, to generate 30 different spectral ranges of light from 9 or fewer reflections by an optical component.

[0005] A photodetector system according to a particular embodiment includes wavelength separators configured to generate light of first, second, and third predetermined spectral ranges from a light source, and first, second, and third photodetector modules configured to receive light of each of the first, second, and third predetermined spectral ranges, wherein the photodetector modules have a plurality of photodetectors and optical components for transmitting light having a predetermined subspectral range to the photodetectors. In a particular example, the wavelength separator is a prism or a diffraction grating. In a particular embodiment, the photodetector system includes three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, one or more photodetector modules optically communicating with each wavelength separator having a plurality of photodetectors, and optical components for transmitting light having a predetermined subspectral range to the photodetectors. In some embodiments, the wavelength separators are configured to transmit light between themselves. The wavelength separators may be positioned along a single plane or along two or more parallel planes. In a particular embodiment, the wavelength separators are positioned in a polygonal configuration, such as a pentagonal or hexagonal configuration. In some embodiments, the wavelength separator is configured to allow light within a predetermined spectral range to pass through. In some embodiments, the wavelength separator is configured to allow light having wavelengths in the range of 200 nm to 1200 nm, such as 360 nm to 960 nm. In some embodiments, each wavelength separator is configured to allow light having a spectral range spanning 75 nm to 150 nm to pass through. In a specific example, each wavelength separator is configured to allow light having a spectral range spanning 100 nm to pass through (for example, light having wavelengths in the range of 360 nm to 460 nm to pass through).

[0006] The photodetector system includes one or more photodetector modules that are optically connected to each wavelength separator. In some embodiments, each photodetector module includes a plurality of photodetectors and optical components configured to transmit light having a predetermined subspectral range to the photodetectors. In some embodiments, each optical component is configured to transmit light having a subspectral range of 5 nm to 50 nm to each photodetector, such as a subspectral range of about 20 nm to each photodetector. The photodetectors and optical components may be positioned within each photodetector module along a single plane or along two or more parallel planes. In certain embodiments, the photodetectors and optical components are positioned within each photodetector module in a polygonal configuration, such as a hexagonal, heptagonal, or octagonal configuration.

[0007] Aspects of the present disclosure also include systems for measuring light from a sample (e.g., in a flow stream). In certain embodiments, the system includes a light source and a clustered wavelength division photodetector system, the clustered wavelength division photodetector system including three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, and one or more photodetector modules optically communicating with each wavelength separator, each having a plurality of photodetectors and optical components for transmitting light having a predetermined subspectral range to the photodetectors. In some embodiments, the system also includes a focusing system for propagating light to the photodetector system. The focusing system may be a free-space optical relay system or may include optical fibers such as optical fiber relay bundles. In some embodiments, the system is a flow cytometer.

[0008] Aspects of the present disclosure also include a method for irradiating a sample in an interrogation field (e.g., in a flow stream) using a light source, collecting and detecting light from the sample using the photodetector system of the present invention, and measuring the detected light at one or more wavelengths. In some embodiments, the light is collected and transmitted to the photodetector system by a free-space optical relay system. In other embodiments, the light is collected and transmitted to the photodetector system by optical fibers, such as an optical fiber relay bundle.

[0009] Kits comprising one or more components of the photodetection system of the present invention are also provided. A kit according to a particular embodiment comprises three or more wavelength separators, a plurality of photodetectors, and optical components. In embodiments, the optical components include collimators, beam splitters, wavelength separators, or combinations thereof. The kit may also include one or more photodetectors, such as photomultiplier tubes (e.g., metal-packaged photomultiplier tubes). [Brief explanation of the drawing]

[0010] [Figure 1A] This figure shows the configuration of a wavelength separator positioned along two parallel planes within a photodetector system according to a specific embodiment. [Figure 1B] This figure shows the configuration of a wavelength separator positioned in a polygonal configuration within a photodetection system according to a specific embodiment. [Figure 2] This figure shows a wavelength separator configured to generate light in first, second, and third spectral ranges according to a specific embodiment. [Figure 3] This figure shows the components of a photodetector module positioned along two parallel axes according to a specific embodiment. [Figure 4] This figure shows the components of a photodetector module positioned in a polygonal configuration according to a specific embodiment. [Figure 5] This figure shows the components of a photodetector module positioned along two parallel planes according to a specific embodiment. [Figure 6] This figure shows a photodetection system having multiple wavelength separators and photodetection modules according to a specific embodiment. [Figure 7A] This figure shows a photodetection system having three or more concentrically arranged wavelength separators optically coupled to a photodetection module according to a specific embodiment, and a photodetection system having three wavelength separators optically coupled to a photodetection module. [Figure 7B]Figure showing a light detection system having three or more wavelength separators arranged concentrically and optically coupled to a light detection module according to a particular embodiment, and showing a light detection system having four wavelength separators optically coupled to the light detection module. [Figure 7C] Figure showing a light detection system having three or more wavelength separators arranged concentrically and optically coupled to a light detection module according to a particular embodiment, and showing a light detection system having five wavelength separators optically coupled to the light detection module. [Figure 7D] Figure showing a light detection system having three or more wavelength separators arranged concentrically and optically coupled to a light detection module according to a particular embodiment, and showing a light detection system having six wavelength separators optically coupled to the light detection module. <好 [Figure 7E] Figure showing a light detection system having three or more wavelength separators arranged concentrically and optically coupled to a light detection module according to a particular embodiment, and showing a three-dimensional view of the light detection system of FIG. 7D. DETAILED DESCRIPTION

[0011] A system for detecting light (e.g., in a frost stream) is described. A light detection system according to a particular embodiment includes a wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, and first, second, and third light detection modules configured to receive each of the light in the first, second, and third predetermined spectral ranges, wherein the light detection module includes a plurality of light detectors and an optical component that transmits light having a predetermined sub-spectral range to the light detectors. Also provided are a system and method for measuring light emitted by a sample (e.g., in a frost stream), and a kit having three or more wavelength separators, a plurality of light detectors, and optical components.

[0012] Before the present invention is described in more detail, it should be understood that the present invention is not limited to the specific embodiments described and is therefore, of course, subject to change. Furthermore, since the scope of the present invention is limited only by the appended claims, it should also be understood that the terms used herein are intended to describe only specific embodiments and are not intended to limit them.

[0013] Where a range of values ​​is provided, unless the context explicitly indicates otherwise, it is understood that each intermediate value between the upper and lower limits of that range, up to one-tenth of the lower limit unit, and any other described values ​​or intermediates within the range described herein are included in the invention. The upper and lower limits of these smaller ranges may independently be included in smaller ranges and are likewise included in the invention, according to any specifically excluded limits in the described range. Where a described range includes one or both limits, ranges excluding either or both of those included limits are also likewise included in the invention.

[0014] Certain ranges are presented herein with a numerical value preceding the term “approximately.” The term “approximately” is used herein to provide literal support for the exact number it precedes, and for any number that the term is close to or approximates the preceding number. In determining whether a number is close to or approximates a specifically stated number, any unstated number that is close to or approximates a specific number may, in the context in which it is presented, provide a substantial equivalent of the specifically stated number.

[0015] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which this invention belongs. Any methods and materials similar or equivalent to those described herein may also be used in carrying out or testing the present invention, but representative exemplary methods and materials are described herein.

[0016] All publications and patents cited herein are incorporated herein by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference, and are incorporated herein by reference to disclose and describe methods and / or materials, and publications are cited in relation to those methods and / or materials. Any citation of a publication relates to its disclosure prior to the filing date, and the present invention should not be construed as acknowledging that such publication has no prior rights on the grounds of prior invention. Furthermore, the publication dates provided may differ from the actual publication dates and may need to be independently verified.

[0017] When used herein and in the appended claims, the articles “a,” “an,” and “the” refer to multiple subjects unless otherwise explicitly indicated by the context. Furthermore, it should be noted that claims may be drafted to exclude any element. Therefore, this statement is intended to serve as a preceding basis for the use of exclusive terms such as “exclusively” and “solely” in connection with the enumeration of elements of the claims or the use of “negative” limitations.

[0018] As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features that can be readily separated from or combined with any of the features of some other embodiments without departing from the scope or spirit of the invention. Any described method may be performed in the order of the described events, or in any other logically possible order.

[0019] Apparatus and methods have been, or will be, described with a functional description for grammatical fluidity, but unless explicitly stated under Section 112 of the United States Patent Act, the claims should not necessarily be interpreted as being limited by a “means” or “step” limitation, but should be granted the full scope of the meaning and equivalents of the definitions provided by the claims under the doctrine of legal equivalents, and if the claims are explicitly stated under Section 112 of the United States Patent Act, they should be granted the full legal equivalents under Section 112 of the United States Patent Act.

[0020] Light detection system Aspects of the present disclosure include clustered wavelength-division photodetector systems configured to detect light from a sample (e.g., light obtained from a flow stream of a flow cytometer). A photodetector system according to a particular embodiment includes wavelength separators configured to generate light of first, second, and third predetermined spectral ranges from a light source, and first, second, and third photodetector modules configured to receive light of each of the first, second, and third predetermined spectral ranges, the photodetector modules having a plurality of photodetectors and optical components for transmitting light having a predetermined subspectral range to the photodetectors. In a particular example, the wavelength separator is a prism or a diffraction grating. In some embodiments, the photodetector system includes three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, one or more photodetector modules optically communicating with each wavelength separator having a plurality of photodetectors, and optical components for transmitting light having a predetermined subspectral range to the photodetectors.

[0021] In an embodiment, light from a sample is split into three or more spectral ranges by passing the light through one or more wavelength separators. Each spectral range of the light generated by the wavelength separator is further split into smaller sub-spectral ranges that are detected by a light detector. In some embodiments, the light detected from the sample is emitted light such as fluorescence. In other embodiments, the light detected from the sample is scattered light. The term "scattered light" is used herein in its conventional sense to refer to the propagation of light energy from particles within the sample that are deflected from the incident beam path (e.g., flowing within a flow stream) by, for example, reflection, refraction, or deflection of the light beam.

[0022] In an embodiment, the light detection system described herein is configured to exhibit little or no light loss from the light collected from the sample. In some embodiments, the light loss due to the transmission of light through the light detection system of interest is 25% or less, such as 20% or less, such as 15% or less, such as 10% or less, such as 5% or less, such as 1% or less, such as 0.5% or less, such as 0.1% or less, such as 0.01% or less, including 0.001% or less. In a particular example, there is no light loss from the light propagating from the sample through the light detection system of the present invention (i.e., it exhibits no measurable light loss). For example, the amount of light from the sample, when transmitted through the light detection system of the present invention, is 1 mW / cm 2 Hereinafter, for example, 0.5 mW / cm 2 Hereinafter, for example, 0.1 mW / cm 2 Hereinafter, for example, 0.05 mW / cm 2 Hereinafter, for example, 0.01 mW / cm 2 Hereinafter, for example, 0.005 mW / cm 2 Hereinafter, for example, 0.001 mW / cm 2 Hereinafter, for example, 0.0005 mW / cm 2 Hereinafter, for example, 0.0001 mW / cm 2 Hereinafter, for example, 0.00005 mW / cm 2 It only decreases hereinafter, including 0.00001 mW / cm 2 Hereinafter.

[0023] As described herein, the photodetector system is configured to generate light in multiple subspectral ranges from light collected from a sample. In some embodiments, light in five or more subspectral ranges, e.g., 10 or more, e.g., 15 or more, e.g., 20 or more, e.g., 25 or more, e.g., 30 or more, e.g., 35 or more, e.g., 40 or more, e.g., 45 or more, and even 50 or more subspectral ranges, is generated from light collected from the sample. In these embodiments, the optical loss shown by the photodetector system and method described herein is 20% or less, e.g., 19% or less, e.g., 18% or less, e.g., 17% or less, e.g., 16% or less, e.g., 15% or less, and includes exhibiting an optical loss of 10% or less. For example, light in five or more subspectral ranges may be generated from light collected from a sample, where the light from the sample, when transmitted through the photodetector system of the present invention, is 1 mW / cm². 2 For example, 0.5 mW / cm² 2 For example, 0.1 mW / cm² 2 For example, 0.05 mW / cm² 2 For example, 0.01 mW / cm² 2 For example, 0.005 mW / cm² 2 For example, 0.001 mW / cm² 2 For example, 0.0005 mW / cm² 2 For example, 0.0001 mW / cm² 2 For example, 0.00005 mW / cm² 2 The decrease was only as follows: 0.00001 mW / cm² 2 Includes the following:

[0024] Light propagating through the photodetector system of the present invention exhibits little to no divergence. In other words, there is little, if any, change to the light beam as it is transmitted through the wavelength separator to the photodetector. In some embodiments, the focal radius of the light transmitted through the photodetector system of the present invention increases by 5% or less, e.g., 4% or less, e.g., 3% or less, e.g., 2% or less, e.g., 1% or less, e.g., 0.5% or less, e.g., 0.1% or less, e.g., 0.01% or less, e.g., 0.001% or less, and includes 0.0001% or less. In certain examples, the focal radius of the light transmitted through the photodetector system of the subject does not increase at all (i.e., it does not show a measurable increase in the focal radius). For example, depending on the size of the light beam transmitted through the photodetector system, the diameter of the light beam increases by 2 mm or less, e.g., 1.5 mm or less, e.g., 1 mm or less, e.g., 0.9 mm or less, e.g., 0.8 mm or less, e.g., 0.7 mm or less, e.g., 0.6 mm or less, e.g., 0.5 mm or less, e.g., 0.4 mm or less, e.g., 0.3 mm or less, e.g., 0.2 mm or less, e.g., 0.1 mm or less, e.g., 0.05 mm or less, e.g., 0.01 mm or less, e.g., 0.001 mm or less, e.g., 0.0001 mm or less, and includes 0.00001 mm or less. In certain examples, the diameter of the light beam does not show a measurable increase when transmitted through the photodetector system (i.e., it increases by 0 mm).

[0025] In some embodiments, the wavelength separator is configured to generate light from three or more predetermined spectral ranges from a light source (for example, light from a sample irradiated with light, as described in detail below), including light from four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, fifteen or more, twenty-five or more, fifty or more, seven-five or more, and one hundred or more predetermined spectral ranges. In a particular example, the photodetection system includes a wavelength separator configured to generate light from a first, second and third predetermined spectral ranges from a light source.

[0026] In some embodiments, the photodetection system includes three or more wavelength separators, e.g., four or more, e.g., five or more, e.g., six or more, e.g., seven or more, e.g., eight or more, e.g., nine or more, e.g., ten or more, e.g., fifteen or more, e.g., twenty-five or more, e.g., fifty or more, e.g., seven-five or more, and one hundred or more wavelength separators. The term “wavelength separator” is used herein in its conventional sense to refer to an optical component configured to separate light collected from a sample into predetermined spectral ranges. In some embodiments, the wavelength separator is configured to separate light collected from a sample into predetermined spectral ranges by allowing light having a predetermined spectral range to pass through and reflecting light from one or more remaining spectral ranges. In other embodiments, the wavelength separator is configured to separate light collected from a sample into predetermined spectral ranges by allowing light having a predetermined spectral range to pass through and absorbing light from one or more remaining spectral ranges. In yet another embodiment, the wavelength separator is configured to spatially diffract the light collected from the sample into predetermined spectral ranges. Each wavelength separator may be any suitable optical separation protocol, such as one or more dichroic mirrors, bandpass filters, diffraction gratings, beam splitters, or prisms. In some embodiments, the wavelength separator is a prism. In other embodiments, the wavelength separator is a diffraction grating. In certain embodiments, the wavelength separator in the optical detection system of the present invention is a dichroic mirror.

[0027] In this embodiment, the wavelength separator is configured to separate the first wavelength X i (in nanometers (nm) units) to the second wavelength X n It is configured to allow light having wavelengths in the range up to nanometers (nm) to pass through. In some embodiments, the wavelength separator is configured to allow light having wavelengths in the range of 100nm to 1500nm, 150nm to 1450nm, 200nm to 1400nm, 250nm to 1350nm, 300nm to 1300nm, 350nm to 1250nm, 400nm to 1200nm, 450nm to 1150nm, 500nm to 1100nm, 550nm to 1050nm, etc. i ~X nIt is configured to allow light having wavelengths in the range of 600 nm to 1000 nm to pass through, and includes allowing light having wavelengths in the range of 600 nm to 1000 nm to pass through. In a particular embodiment, the wavelength separator in the photodetector system of the subject is configured to allow light having wavelengths in the range of 360 nm to 960 nm to pass through.

[0028] In this embodiment, each of the target wavelength separators emits light X in a predetermined spectral range. s It is configured to generate (in nanometer (nm) units). The predetermined spectral range may vary, and in certain embodiments, the wavelength separator in question is configured to generate light (X) in spectral ranges such as 50 nm to 300 nm, e.g., 75 nm to 275 nm, e.g., 100 nm to 250 nm, e.g., 125 nm to 225 nm, and 150 nm to 200 nm. s ) is configured to generate ). In a particular embodiment, each wavelength separator is configured to generate 100 nm (i.e., X s It is configured to generate light with a spectral range extending to 100 nm.

[0029] In one example, the photodetection system includes a wavelength separator configured to generate light in a first predetermined spectral range of 360 nm to 480 nm, light in a second predetermined spectral range of 480 nm to 600 nm, light in a third predetermined spectral range of 600 nm to 720 nm, light in a fourth predetermined spectral range of 720 nm to 840 nm, and light in a fifth predetermined spectral range of 840 nm to 960 nm.

[0030] In another example, the photodetector system operates in the 360nm-480nm range (i.e., X s It includes a first wavelength separator configured to allow light having wavelengths in the range of 120 nm to pass through, a second wavelength separator configured to allow light having wavelengths in the range of 480 nm to 600 nm to pass through, a third wavelength separator configured to allow light having wavelengths in the range of 600 nm to 720 nm to pass through, a fourth wavelength separator configured to allow light having wavelengths in the range of 720 nm to 840 nm to pass through, and a fifth wavelength separator configured to allow light having wavelengths in the range of 840 nm to 960 nm to pass through.

[0031] In some embodiments, the photodetector system in question includes three or more wavelength separators that are optically in communication with one another, such as being positioned to transmit light between them. The wavelength separators may be oriented relative to each other in the photodetector system (as referenced in the XZ plane) at angles including 45° to 90°, such as 10° to 180°, e.g., 15° to 170°, e.g., 20° to 160°, e.g., 25° to 150°, e.g., 30° to 120°. In some cases, the wavelength separators are positioned along a single plane. In other examples, the wavelength separators are positioned along two or more planes. For example, the wavelength separators may be positioned along two or more parallel planes, e.g., three or more, e.g., four or more, or even five or more parallel planes. In certain examples, the wavelength separator is arranged in a geometric configuration, and the configurations in question include, but are not limited to, square, rectangular, trapezoidal, triangular, hexagonal, heptagonal, octagonal, nonagonal, decagonal, dodecagonal, circular, elliptical, and irregular configurations. In certain embodiments, the wavelength separator is arranged in a pentagonal configuration. In other embodiments, the wavelength separator is arranged in a hexagonal configuration.

[0032] In some embodiments, wavelength separators are configured to transmit light between them. In some examples, each wavelength separator is configured to allow light from one spectral range to pass through and transmit light from one or more remaining spectral ranges to another wavelength separator (e.g., by reflection). In one example, the photodetector system includes three wavelength separators. The first wavelength separator is configured to receive light from a sample, allow light from a first spectral range to pass through, and transmit light from a second spectral range to a second wavelength separator. The second wavelength separator is configured to allow light from a third spectral range to pass through and transmit light from a fourth spectral range to a third wavelength separator. In some cases, the light from the third spectral range is a portion of the light from the second spectral range, for example, 90% or less, e.g., 85% or less, e.g., 80% or less, e.g., 75% or less, e.g., 70% or less, e.g., 65% or less, e.g., 60% or less, e.g., 55% or less, e.g., 50% of the light from the second spectral range. The third wavelength separator is configured to allow light of the fifth spectral range to pass through. In some cases, the light of the fifth spectral range is a portion of the light of the fourth spectral range, for example, 90% or less, for example, 85% or less, for example, 80% or less, for example, 75% or less, for example, 70% or less, for example, 65% or less, for example, 60% or less, for example, 55% or less, for example, 50% of the light of the fourth spectral range.

[0033] In another example, the photodetector system includes five wavelength separators. The first wavelength separator is configured to receive light from a sample, pass light in a first spectral range through it, and transmit light in a second spectral range to the second wavelength separator. The second wavelength separator is configured to pass light in a third spectral range through it and transmit light in a fourth spectral range to the third wavelength separator. In some cases, the light in the third spectral range is a portion of the light in the second spectral range, for example, 90% or less of the light in the second spectral range, e.g., 85% or less, e.g., 80% or less, e.g., 75% or less, e.g., 70% or less, e.g., 65% or less, e.g., 60% or less, e.g., 55% or less, e.g., 50% or less. The third wavelength separator is configured to pass light in a fifth spectral range through it and transmit light in a sixth spectral range to the fourth wavelength separator. In some cases, the light in the fifth spectral range is a portion of the light in the fourth spectral range, for example, a spectral range that accounts for 90% or less of the light in the fourth spectral range, for example, 85% or less, for example, 80% or less, for example, 75% or less, for example, 70% or less, for example, 65% or less, for example, 60% or less, for example, 55% or less, for example, 50%. The fourth wavelength separator is configured to allow the light in the seventh spectral range to pass through and to transmit the light in the eighth spectral range to the fifth wavelength separator. In some examples, the light in the seventh spectral range is a portion of the light in the sixth spectral range, for example, a spectral range that accounts for 90% or less of the light in the sixth spectral range, for example, 85% or less, for example, 80% or less, for example, 75% or less, for example, 70% or less, for example, 65% or less, for example, 60% or less, for example, 55% or less, for example, 50%. The fifth wavelength separator is configured to allow the light in the ninth spectral range to pass through. In some examples, the ninth spectral range of light is a portion of the eighth spectral range of light, for example, less than 90%, less than 85%, less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 55%, or less than 50% of the eighth spectral range of light.

[0034] In a particular embodiment, the photodetection system includes five wavelength separators configured to separate light having wavelengths in the range of 360 nm to 960 nm, wherein the first wavelength separator is configured to allow light having wavelengths in the range of 360 nm to 480 nm to pass through and transmit light having wavelengths in the range of 480 nm to 960 nm to the second wavelength separator, the second wavelength separator is configured to allow light having wavelengths in the range of 480 nm to 600 nm to pass through and transmit light having wavelengths in the range of 600 nm to 960 nm to the third wavelength separator, the third wavelength separator is configured to allow light having wavelengths in the range of 600 nm to 720 nm to pass through and transmit light having wavelengths in the range of 720 nm to 960 nm to the fourth wavelength separator, and the fourth wavelength separator is configured to allow light having wavelengths in the range of 720 nm to 840 nm to pass through and transmit light having wavelengths in the range of 840 nm to 960 nm to the fifth wavelength separator. In this embodiment, the fifth wavelength separator is configured to allow light having wavelengths in the range of 840 nm to 960 nm to pass through.

[0035] Figure 1A shows the configuration of wavelength separators positioned along two parallel planes within a photodetector system according to a particular embodiment. Light from a sample is transmitted through a focusing system OC having optical components configured to pass light having a set of wavelengths 101 (e.g., 360 nm to 960 nm) to a first wavelength separator WS1, which is configured to pass light SR1 (e.g., 360 nm to 480 nm) in a first spectral range and transmit light having a set of wavelengths 102 (e.g., 480 nm to 960 nm) to a second wavelength separator WS2. The second wavelength separator WS2 is configured to pass light SR2 (e.g., 480 nm to 600 nm) in a second spectral range and transmit light having a set of wavelengths 103 (e.g., 600 nm to 960 nm) to a third wavelength separator WS3. The third wavelength separator WS3 is configured to pass light SR3 (e.g., 600 nm to 720 nm) in a third spectral range and transmit light having a set of wavelengths 104 (e.g., 720 nm to 960 nm) to the fourth wavelength separator WS4. The fourth wavelength separator WS4 is configured to pass light SR4 (e.g., 720 nm to 840 nm) in a fourth spectral range and transmit light having a fifth spectral range SR5 (e.g., 840 nm to 960 nm). In this embodiment, the light is transmitted along a zigzag optical path. Each of the spectral ranges of light SR1, SR2, SR3, SR4, and SR5 is transmitted to one or more photodetection modules (as will be described in more detail below).

[0036] Figure 1B shows the configuration of a polygonal-positioned wavelength separator in a photodetection system according to a specific embodiment. Light from a sample is transmitted through a focusing system OC having optical components configured to pass a set of light with wavelengths 201 (e.g., 200 nm to 1200 nm) to a first wavelength separator WS1. The first wavelength separator WS1 is configured to pass light SR1 (e.g., 200 nm to 400 nm) in a first spectral range and transmit light with wavelengths 202 (e.g., 400 nm to 1200 nm) to a second wavelength separator WS2. The second wavelength separator WS2 is configured to pass light SR2 (e.g., 400 nm to 600 nm) in a second spectral range and transmit light with wavelengths 203 (e.g., 600 nm to 1200 nm) to a third wavelength separator WS3. The third wavelength separator WS3 is configured to pass light SR3 (e.g., 600 nm to 800 nm) in a third spectral range and transmit light having a set of wavelengths 204 (e.g., 800 nm to 1200 nm) to the fourth wavelength separator WS4. The fourth wavelength separator WS4 is configured to pass light SR4 (e.g., 800 nm to 1000 nm) in a fourth spectral range and transmit light having a fifth spectral range SR5 (e.g., 1000 nm to 1200 nm). In this embodiment, the light is transmitted along a star-shaped optical path. Each of the spectral ranges of light SR1, SR2, SR3, SR4, and SR5 is transmitted to one or more photodetection modules (as will be described in more detail below).

[0037] Figure 2 shows a wavelength separator configured to generate light in first, second, and third spectral ranges according to a particular embodiment. Light from a sample is transmitted through a focusing system OC having optical components configured to pass through a set of wavelengths (e.g., 200 nm to 1200 nm) to a wavelength separator WS configured to generate light in the first spectral range SR1, light in the second spectral range SR2, and light in the third spectral range SR3. Each of SR1, SR2, and SR3 is transmitted to the first, second, and third photodetector modules, respectively (as will be described in more detail below).

[0038] As summarized above, a photodetection system includes one or more photodetection modules configured to receive light in a predetermined spectral range generated by one or more wavelength separators. In some embodiments, the photodetection system includes wavelength separators configured to generate light in first, second, and third predetermined spectral ranges, and first, second, and third photodetection modules configured to receive light in each of the first, second, and third predetermined spectral ranges.

[0039] In some embodiments, the photodetection system includes a photodetection module that is in optical communication with each wavelength separator. In some embodiments, the photodetection module is positioned in physical contact with the wavelength separator, such as when an aperture to the photodetection module is physically coupled to the wavelength separator. In other embodiments, each photodetection module is positioned at a distance of 0.001 mm or more from the wavelength separator, for example, 0.005 mm or more, for example, 0.01 mm or more, for example, 0.05 mm or more, for example, 0.1 mm or more, for example, 0.5 mm or more, for example, 1 mm or more, for example, 2 mm or more, for example, 3 mm or more, for example, 4 mm or more, for example, 5 mm or more, for example, 10 mm or more, for example, 15 mm or more, for example, 25 mm or more, and even 50 mm or more. For example, each photodetector module may be positioned at a distance of 0.0001mm to 100mm from the wavelength separator, for example, 0.0005mm to 95mm, for example, 0.001mm to 90mm, for example, 0.005mm to 85mm, for example, 0.01mm to 80mm, for example, 0.05mm to 75mm, for example, 0.1mm to 70mm, for example, 0.5mm to 65mm, for example, 1mm to 60mm, for example, 1.5mm to 55mm, or even 2mm to 50mm.

[0040] The photodetector module may be removably connected to the wavelength separator. The term “removable” is used herein in its conventional sense to mean that each photodetector module or wavelength separator can be freely removed and reattached. The photodetector module or wavelength separator may be connected by any preferred protocol. In certain embodiments, the photodetector module and wavelength separator are connected together by fasteners such as hook-and-loop fasteners, magnets, latches, notches, countersunk holes, counterbores, grooves, pins, tethers, hinges, Velcro, non-permanent adhesives, or combinations thereof. In certain examples, the photodetector module is connected to the wavelength separator by slotting the wavelength separator into a groove in the photodetector module. In yet another example, the wavelength separator is connected to the photodetector module by one or more screws.

[0041] In some embodiments, light from each wavelength separator is transmitted to each photodetector module by a focusing system. Each focusing system may be any suitable focusing protocol that collects light across the spectral range that has passed through the wavelength separator and directs that light to the photodetector module. In some embodiments, the focusing system includes optical fibers, such as optical fiber relay bundles. In other embodiments, the focusing system is a free-space optical relay system.

[0042] In some embodiments, each focusing system may be physically bonded to a photodetector module using an adhesive or the like, co-molded with each photodetector module, or integrated into each photodetector module. In certain embodiments, each photodetector module and focusing system are integrated into a single unit. In some examples, each photodetector module is bonded to the focusing system using connectors that fasten the focusing system to each photodetector module, such as hook-and-loop fasteners, magnets, latches, notches, countersunk holes, counterbores, grooves, pins, tethers, hinges, Velcro, non-permanent adhesives, or combinations thereof.

[0043] In other embodiments, each light detection module and the focusing system are in optical communication but not in physical contact. In embodiments, the focusing system may be positioned at a distance of 0.001 mm or more from the light detection module, for example, 0.005 mm or more, for example, 0.01 mm or more, for example, 0.05 mm or more, for example, 0.1 mm or more, for example, 0.5 mm or more, for example, 1 mm or more, for example, 10 mm or more, for example, 25 mm or more, for example, 50 mm or more, and 100 mm or more from the light detection module.

[0044] In certain embodiments, the focusing system includes optical fibers. For example, the focusing system may be an optical fiber relay bundle, and light of a spectral range that has passed through a wavelength separator is transmitted through the optical fiber relay bundle to a photodetector module. Any optical fiber relay system may be employed to transmit light, and in certain embodiments, preferred optical fiber relay systems include, but are not limited to, those described in U.S. Patent No. 6,809,804, the disclosure of which is incorporated herein by reference.

[0045] In other embodiments, each focusing system is a free-space light relay system. The term “free-space light relay” is used herein in its conventional sense to refer to light propagation that uses the configuration of one or more optical components to direct light of a spectral range passed through a wavelength separator through free space to a photodetector module. In certain embodiments, the free-space light relay system includes a housing having a proximal end and a distal end, the proximal end being coupled to the photodetector module. The free-space relay system may include any combination of different optical components, such as one or more of lenses, mirrors, slits, pinholes, wavelength separators, or combinations thereof. For example, in some embodiments, the free-space light relay system in question includes one or more focusing lenses. In other embodiments, the free-space light relay system of the present invention includes one or more mirrors. In yet another embodiment, the free-space light relay system includes a collimating lens. Certain embodiments of suitable free-space optical relay systems for propagating light across a spectral range from a wavelength separator include, but are not limited to, optical relay systems described in U.S. Patents No. 7,643,142, No. 7,728,974, and No. 8,223,445, the disclosures of which are incorporated herein by reference.

[0046] The photodetector modules may be arranged in any geometric configuration in the photodetector system of the present invention as needed (for example, they may be mounted together). The photodetector modules may be arranged along one or more planes. In some embodiments, the photodetector modules may be oriented relative to each other at angles in the range including 0° to 180°, e.g., 10° to 170°, e.g., 20° to 160°, e.g., 25° to 150°, e.g., 30° to 120°, and 45° to 90° (as referred to in the XZ plane). In embodiments, the photodetector modules may be arranged relative to each other at the same or different angles, depending on the number of photodetector modules in the photodetector system. For example, in a particular example, the angle between the first photodetector module and the second photodetector module is the same as the angle between the second photodetector module and the third photodetector module. In some embodiments, the angle between the first photodetector module and the second photodetector module is different from the angle between the second photodetector module and the third photodetector module. In some embodiments, the light detection module is positioned in a geometric arrangement such as a star configuration, triangular configuration, square configuration, rectangular configuration, trapezoidal configuration, triangular configuration, hexagonal configuration, heptagonal configuration, octagonal configuration, nonagonal configuration, decagonal configuration, dodecagonal configuration, circular configuration, elliptical configuration, and irregular shape configuration.

[0047] In some embodiments, each photodetector module includes an optical tuning component configured to transmit light having a predetermined subspectral range to one or more photodetectors. “Optical tuning” means that the light is modified or tuned when transmitted to each photodetector within the photodetector module. In some embodiments, optical tuning includes propagating light having a predetermined subspectral range to the photodetectors. In some embodiments, each photodetector module includes one or more optical tuning components configured to separate the light transmitted from a wavelength separator into a predetermined subspectral range by allowing light having a predetermined subspectral range to pass through and reflecting light from one or more remaining spectral ranges. In other embodiments, the optical tuning component is configured to separate the light transmitted from a wavelength separator into a predetermined subspectral range by allowing light having a predetermined subspectral range to pass through and absorbing light from one or more remaining spectral ranges. In yet another embodiment, the optical tuning component is configured to spatially diffract the light transmitted from the wavelength separator into a predetermined subspectral range. The optical tuning component may be any preferred optical separation protocol, such as one or more dichroic mirrors, bandpass filters, diffraction gratings, beam splitters, or prisms. In certain embodiments, the optical tuning component within the photodetector module, configured to separate light transmitted from a wavelength separator into a predetermined subspectral range, is a dichroic mirror.

[0048] (As described above) Depending on the wavelength of light passed from the wavelength separator to the photodetector module, one or more optical components within the photodetector module will have a first wavelength Y i (nanometer (nm)) (Units) From the second wavelength Y nThey can be configured to transmit light having wavelengths in the range of nanometers (nm) to a photodetector. In some embodiments, one or more optical components are configured to transmit light having wavelengths in the range of 100 nm to 1500 nm to a photodetector, such as 150 nm to 1450 nm, 200 nm to 1400 nm, 250 nm to 1350 nm, 300 nm to 1300 nm, 350 nm to 1250 nm, 400 nm to 1200 nm, 450 nm to 1150 nm, 500 nm to 1100 nm, and 550 nm to 1050 nm, and include propagating light having wavelengths in the range of 600 nm to 1000 nm to a photodetector.

[0049] In this embodiment, the optical components within each photodetector module are configured to meet a predetermined subspectral range Y of light. s The optical components are configured to transmit light (in nanometers (nm) units) to each photodetector. The predetermined subspectral range transmitted by each optical component may vary, and a particular optical component in question may be configured to transmit light in a subspectral range including 5nm to 50nm, for example, 6nm to 49nm, for example, 7nm to 48nm, for example, 8nm to 47nm, for example, 9nm to 46nm, and including 10nm to 45nm. In a particular embodiment, the optical component is configured to transmit light in a spectral range extending to 20nm.

[0050] For example, in one example, one or more optical components are 20 nm (i.e., Y s In the subspectral range extending to 20 nm, 360 nm (i.e., Y i =360nm)~480nm(that is, Y nThe optical module is configured to allow light having wavelengths in the range of 360nm to 480nm to pass through. In this embodiment, the optical module includes a first optical component configured to transmit light having wavelengths in the range of 360nm to 380nm to the photodetector, a second optical component configured to transmit light having wavelengths in the range of 380nm to 400nm to the photodetector, a third optical component configured to transmit light having wavelengths in the range of 400nm to 420nm to the photodetector, a fourth optical component configured to transmit light having wavelengths in the range of 420nm to 440nm to the photodetector, a fifth optical component configured to transmit light having wavelengths in the range of 440nm to 460nm to the photodetector, and a sixth optical component configured to transmit light having wavelengths in the range of 460nm to 480nm to the photodetector.

[0051] In some embodiments, the optical components within each photodetector module are optically connected to one another, such as being positioned to transmit light between them. The optical components may be oriented in the photodetector module (as referenced in the XZ plane) at angles including 10° to 180°, e.g., 15° to 170°, e.g., 20° to 160°, e.g., 25° to 150°, e.g., 30° to 120°, and 45° to 90° relative to each other. In some examples, the optical components are positioned along a single plane. In other examples, the optical components are positioned along two or more planes. For example, the optical components may be positioned along two or more parallel planes, e.g., three or more, e.g., four or more, and five or more parallel planes. In certain examples, the optical components are arranged in a geometric configuration, and the configurations in question include, but are not limited to, square, rectangular, trapezoidal, triangular, hexagonal, heptagonal, octagonal, nonagonal, decagonal, dodecagonal, circular, elliptical, and irregular configurations. In certain embodiments, the optical components are arranged in a hexagonal configuration. In other embodiments, the optical components are arranged in a heptagonal configuration.

[0052] In some embodiments, optical components are configured to transmit light between them. In some examples, each optical component is configured to pass light in a certain spectral range and transmit light in one or more remaining spectral ranges to another optical component (e.g., by reflection). In one example, a photodetector module includes three optical components. A first optical component is configured to receive light from a wavelength separator, pass light in a first subspectral range, and transmit light in a second subspectral range to a second optical component. A second optical component is configured to pass light in a third subspectral range and transmit light in a fourth subspectral range to a third optical component. In some examples, the light in the third subspectral range is a subspectral range that is a portion of the light in the second subspectral range, for example, 90% or less, e.g., 85% or less, e.g., 80% or less, e.g., 75% or less, e.g., 70% or less, e.g., 65% or less, e.g., 60% or less, e.g., 55% or less, e.g., 50% or less. The third optical component is configured to allow light in the fifth subspectral range to pass through. In some examples, the light in the fifth subspectral range is a subspectral range that is a portion of the light in the fourth subspectral range, for example, 90% or less, e.g., 85% or less, e.g., 80% or less, e.g., 75% or less, e.g., 70% or less, e.g., 65% or less, e.g., 60% or less, e.g., 55% or less, e.g., 50% of the light in the fourth subspectral range.

[0053] In another example, a photodetector module includes five optical components. The first optical component is configured to receive light from a wavelength separator, pass light in a first subspectral range through it, and transmit light in a second subspectral range to the second optical component. The second optical component is configured to pass light in a third subspectral range through it and transmit light in a fourth subspectral range to the third optical component. In some examples, the third subspectral range of light is a subspectral range that includes a portion of the light in the second subspectral range, for example, 90% or less, e.g., 85% or less, e.g., 80% or less, e.g., 75% or less, e.g., 70% or less, e.g., 65% or less, e.g., 60% or less, e.g., 55% or less, e.g., 50% of the light in the second subspectral range. The third optical component is configured to pass light in a fifth subspectral range through it and transmit light in a sixth subspectral range to the fourth optical component. In some examples, the light in the fifth subspectral range is a subspectral range that includes a portion of the light in the fourth subspectral range, for example, less than 90%, for example less than 85%, for example less than 80%, for example less than 75%, for example less than 70%, for example less than 65%, for example less than 60%, for example less than 55%, for example less than 50% of the light in the fourth subspectral range. The fourth optical component is configured to allow the light in the seventh subspectral range to pass through and to transmit the light in the eighth subspectral range to the fifth optical component. In some examples, the light in the seventh spectral range is a subspectral range that includes a portion of the light in the sixth spectral range, for example, less than 90%, for example less than 85%, for example less than 80%, for example less than 75%, for example less than 70%, for example less than 65%, for example less than 60%, for example less than 55%, for example less than 50% of the light in the sixth spectral range. The fifth optical component is configured to allow the light in the ninth subspectral range to pass through. In some examples, the ninth subspectral range of light is a subspectral range that comprises a portion of the eighth subspectral range of light, for example, less than 90%, less than 85%, less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 55%, or less than 50% of the eighth subspectral range of light.

[0054] Figure 3 shows the components of a photodetector module positioned along two parallel planes according to a particular embodiment. Light SRx from a wavelength separator (as described above) having a set of wavelengths 301 (e.g., 360 nm to 480 nm) is transmitted to a first optical component OA1, which is configured to pass light sSR1 (e.g., 360 nm to 380 nm) in a first subspectral range and transmit light having a set of wavelengths 302 (e.g., 380 nm to 480 nm) to a second optical component OA2. Light sSR1 in the first subspectral range is transmitted to a first photodetector D1. The second optical component OA2 is configured to pass light sSR2 (e.g., 380 nm to 400 nm) in a second subspectral range and transmit light having a set of wavelengths 303 (e.g., 400 nm to 480 nm) to a third optical component OA3. Light sSR2 in the second subspectral range is transmitted to a second photodetector D2. The third optical component OA3 is configured to allow light sSR3 (e.g., 400 nm to 420 nm) in a third subspectral range to pass through, and to transmit light having a set of wavelengths 304 (e.g., 420 nm to 480 nm) to the fourth optical component OA4. The light sSR3 in the third subspectral range is transmitted to the third photodetector D3. The fourth optical component OA4 is configured to allow light sSR4 (e.g., 420 nm to 440 nm) in a fourth subspectral range to pass through, and to transmit light having a set of wavelengths 305 (e.g., 440 nm to 480 nm) to the fifth optical component OA5. The light sSR4 in the fourth subspectral range is transmitted to the fourth photodetector D4. The fifth optical component OA5 is configured to allow light sSR5 in the fifth subspectral range (e.g., 440 nm to 460 nm) to pass through, and to transmit light having a set of wavelengths 306 (e.g., 460 nm to 480 nm) to the sixth optical component OA6. The light sSR5 in the fifth subspectral range is transmitted to the fifth photodetector D5. The sixth optical component OA6 is configured to allow light sSR6 in the sixth subspectral range (e.g., 460 nm to 480 nm) to pass through. The light sSR6 in the sixth subspectral range is transmitted to the sixth photodetector D6. In this embodiment, the light is transmitted sequentially along the zigzag optical path.

[0055] Figure 4 shows the components of a photodetector module positioned in a polygonal configuration according to a specific embodiment. Light SRx from a wavelength separator (as described above) having a set of wavelengths 401 (e.g., 200 nm to 500 nm) is transmitted to a first optical component OA1, which is configured to pass light sSR1 (e.g., 200 nm to 250 nm) in a first subspectral range and transmit light having a set of wavelengths 402 (e.g., 250 nm to 500 nm) to a second optical component OA2. Light sSR1 in the first subspectral range is transmitted to a first photodetector D1. The second optical component OA2 is configured to pass light sSR2 (e.g., 250 nm to 300 nm) in a second subspectral range and transmit light having a set of wavelengths 403 (e.g., 300 nm to 500 nm) to a third optical component OA3. Light sSR2 in the second subspectral range is transmitted to a second photodetector D2. The third optical component OA3 is configured to allow light sSR3 (e.g., 300nm to 350nm) in a third subspectral range to pass through, and to transmit light having a set of wavelengths 404 (e.g., 350nm to 500nm) to the fourth optical component OA4. The light sSR3 in the third subspectral range is transmitted to the third photodetector D3. The fourth optical component OA4 is configured to allow light sSR4 (e.g., 350nm to 400nm) in a fourth subspectral range to pass through, and to transmit light having a set of wavelengths 405 (e.g., 400nm to 500nm) to the fifth optical component OA5. The light sSR4 in the fourth subspectral range is transmitted to the fourth photodetector D4. The fifth optical component OA5 is configured to allow light sSR5 (e.g., 400 nm to 450 nm) in the fifth subspectral range to pass through, and to transmit light having a set of wavelengths 406 (e.g., 450 nm to 500 nm) to the sixth optical component OA6. The light sSR5 in the fifth subspectral range is transmitted to the fifth photodetector D5. The sixth optical component OA6 is configured to allow light sSR6 (e.g., 450 nm to 500 nm) in the sixth subspectral range to pass through. The light sSR6 in the sixth subspectral range is transmitted to the sixth photodetector D6. In this embodiment, the light is transmitted along a star-shaped optical path.

[0056] Figure 5 shows the components of a photodetector module positioned along two parallel planes according to a particular embodiment. Light SRx from a wavelength separator (as described above) having a set of wavelengths 501 (e.g., 360 nm to 600 nm) is transmitted to a first optical component OA1, which is configured to pass light sSR1 (e.g., 360 nm to 400 nm) in a first subspectral range and transmit light having a set of wavelengths 502 (e.g., 400 nm to 600 nm) to a second optical component OA2. Light sSR1 in the first subspectral range is transmitted to a first photodetector D1. The second optical component OA2 is configured to pass light sSR2 (e.g., 400 nm to 440 nm) in a second subspectral range and transmit light having a set of wavelengths 503 (e.g., 440 nm to 600 nm) to a third optical component OA3. Light sSR2 in the second subspectral range is transmitted to a second photodetector D2. The third optical component OA3 is configured to allow light sSR3 in a third subspectral range (e.g., 440 nm to 480 nm) to pass through and transmit light having a set of wavelengths 504 (e.g., 480 nm to 600 nm) to the fourth optical component OA4. The light sSR3 in the third subspectral range is transmitted to the third photodetector D3. The fourth optical component OA4 is configured to allow light sSR4 in a fourth subspectral range (e.g., 480 nm to 520 nm) to pass through and transmit light having a set of wavelengths 505 (e.g., 520 nm to 600 nm) to the fifth optical component OA5. The light sSR4 in the fourth subspectral range is transmitted to the fourth photodetector D4. The fifth optical component OA5 is configured to allow light sSR5 (e.g., 520 nm to 560 nm) in the fifth subspectral range to pass through, and to transmit light having a set of wavelengths 506 (e.g., 560 nm to 600 nm) to the sixth optical component OA6. The light sSR5 in the fifth subspectral range is transmitted to the fifth photodetector D5. The sixth optical component OA6 is configured to allow light sSR6 (e.g., 560 nm to 600 nm) in the sixth subspectral range to pass through. The light sSR6 in the sixth subspectral range is transmitted to the sixth photodetector D6. In this embodiment, the light is transmitted along a round-trip zigzag optical path.

[0057] As described above, the photodetector system is configured to generate light in multiple subspectral ranges from light collected from a sample. In some embodiments, the photodetector system is configured to generate two or more distinct spectral ranges of light for each reflection by an optical component (e.g., a dichroic mirror) within the photodetector system, e.g., three or more, e.g., four or more, and to generate five or more distinct spectral ranges of light for each reflection by an optical component within the photodetector system of the present invention. In certain embodiments, the photodetector system is configured to generate 30 distinct spectral ranges using 10 or fewer reflections by an optical component, e.g., generating 30 distinct spectral ranges of light from 9 or fewer reflections by an optical component. In certain examples, the ratio of the generated distinct spectral ranges to the number of reflections by the optical component within the photodetector system of the subject may be in the range of 2:1 to 10:1, including 3:1 to 5:1, e.g., 3:1 to 7:1. In some examples, the optical component is configured to collimate light. The term "collimate" is used in its traditional sense to refer to optically adjusting the collinearity of light propagation, or reducing the divergence of light from a common propagation axis. In some examples, collimating includes narrowing the spatial cross-section of a light beam. In other examples, collimating includes changing the direction of a light beam, for example, changing the propagation of the light beam by 1° or more, e.g., 5° or more, e.g., 10° or more, e.g., 15° or more, e.g., 20° or more, e.g., 25° or more, e.g., 30° or more, e.g., 45° or more, e.g., 60° or more, e.g., 75° or more, and changing the direction of light propagation by 90° or more. In yet other examples, collimating is a reduction protocol for reducing the dimensions of light (e.g., a beam spot), for example, reducing the dimensions by 5% or more, 10% or more, 25% or more, 50% or more, and 75% or more.

[0058] Light in each subspectral range is transmitted to the photodetector by an optical component. In some embodiments, the optical component is in physical contact with the photodetector. In other embodiments, the optical component is in optical communication with the active surface of the photodetector and may be positioned at a distance of 0.001 mm or more from the photodetector, e.g., 0.005 mm or more, e.g., 0.01 mm or more, e.g., 0.05 mm or more, e.g., 0.1 mm or more, e.g., 0.5 mm or more, e.g., 1 mm or more, e.g., 10 mm or more, e.g., 25 mm or more, e.g., 50 mm or more, and 100 mm or more from the photodetector.

[0059] The photodetector may be removably connected to each optical component within the photodetector module of the subject. The photodetector and optical components may be connected by any preferred protocol. In certain embodiments, the photodetector and optical components are connected together by co-mounting the photodetector with the optical components or with fasteners such as hook-and-loop fasteners, magnets, latches, notches, countersunk holes, counterbores, grooves, pins, tethers, hinges, Velcro, non-permanent adhesives, or combinations thereof. In certain examples, the photodetector is connected to the optical components by slotting the wavelength separator into a groove in the photodetector module. In yet another example, the photodetector is connected to the optical components by one or more screws.

[0060] In some embodiments, each photodetector module includes two or more photodetectors, for example, three or more, for example, four or more, for example, five or more, for example, six or more, for example, seven or more, for example, eight or more, for example, nine or more, for example, ten or more, for example, fifteen or more, for example, twenty-five or more, for example, fifty or more, and one hundred or more photodetectors. In some embodiments, the photodetector module includes one or more photodetector arrays. The term “photodetector array” is used in its conventional sense to refer to an arrangement or series of two or more photodetectors. In some embodiments, a photodetector array may include two or more photodetectors, for example, three or more photodetectors, for example, four or more photodetectors, for example, five or more photodetectors, for example, six or more photodetectors, for example, seven or more photodetectors, for example, eight or more photodetectors, for example, nine or more photodetectors, for example, ten or more photodetectors, for example, twelve or more photodetectors, and may include fifteen or more photodetectors. The photodetectors within each array may be arranged in any geometric configuration as desired, and the configurations include, but are not limited to, square, rectangular, trapezoidal, triangular, hexagonal, heptagonal, octagonal, nonagonal, decagonal, dodecagonal, circular, elliptical, and irregular configurations. The photodetectors within each photodetector array may be oriented relative to the other at angles including 45° to 90°, such as 10° to 180°, e.g., 15° to 170°, e.g., 20° to 160°, e.g., 25° to 150°, e.g., 30° to 120° (as referenced in the XZ plane).

[0061] The photodetector may be any suitable optical sensor, including, among other types of photodetectors, active pixel sensors (APS), avalanche photodiodes, image sensors, charge-coupled devices (CCDs), enhanced charge-coupled devices (ICCDs), complementary metal-oxide-semiconductor (CMOS) image sensors or N-type metal-oxide-semiconductor (NMOS) image sensors, light-emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photocells, photodiodes, photomultiplier tubes, phototransistors, quantum dot photoconductors or photodiodes, and combinations thereof. In certain embodiments, the photodetector includes a photomultiplier tube, such as a metal-packaged photomultiplier tube.

[0062] The photodetector in question is configured to measure light collected at one or more wavelengths, e.g., two or more wavelengths, e.g., five or more different wavelengths, e.g., ten or more different wavelengths, e.g., 25 or more different wavelengths, e.g., fifty or more different wavelengths, e.g., 100 or more different wavelengths, e.g., 200 or more different wavelengths, e.g., 300 or more different wavelengths, and includes measuring light emitted by a sample in a flow stream at 400 or more different wavelengths.

[0063] In the embodiments, the photodetector is configured to measure light continuously or at discrete intervals. In some examples, the photodetector in question is configured to measure the collected light continuously. In other examples, the photodetector is configured to measure light at discrete intervals, such as every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds (including every 1000 milliseconds), or some other interval.

[0064] Figure 6 shows a photodetection system having multiple wavelength separators and photodetection modules according to a specific embodiment. Light from a sample is transmitted through a focusing system OC having optical components configured to pass light having a set of wavelengths 601 to a first wavelength separator WS1. The first wavelength separator WS1 is configured to pass light SR1 of a first spectral range and transmit light having a set of wavelengths 602 to a second wavelength separator WS2. Light of spectral range SR1 is transmitted to a first photodetection module LDM1. The second wavelength separator WS2 is configured to pass light SR2 of a second spectral range and transmit light having a set of wavelengths 603 to a third wavelength separator WS3. Light of spectral range SR2 is transmitted to a second photodetection module LDM2. The third wavelength separator WS3 is configured to pass light SR3 of a third spectral range and transmit light having a set of wavelengths 604 to a fourth wavelength separator WS4. Light in spectral range SR3 is transmitted to the third photodetector module LDM3. The fourth wavelength separator WS4 is configured to allow light in the fourth spectral range SR4 to pass through and transmit light having light in the fifth spectral range SR5 to the fifth photodetector module LDM5. Light in spectral range SR4 is transmitted to the fourth photodetector module LDM4. In this embodiment, each of the spectral ranges SR1, SR2, SR3, SR4, and SR5 of light is transmitted to the photodetector modules LDM1, LDM2, LDM3, LDM4, and LDM5, respectively, and these photodetector modules have a front-to-back zigzag configuration as described in Figure 5 above.

[0065] Figure 7 shows a photodetector system according to a particular embodiment, having three or more concentrically arranged wavelength separators optically coupled to a photodetector module. Figure 7A shows a photodetector system having three wavelength separators 711a, 711b, and 711c. Each wavelength separator is optically coupled to photodetector modules 712a, 712b, and 712c. Figure 7B shows a photodetector system having four wavelength separators 721a, 721b, 721c, and 721d. Each wavelength separator is optically coupled to photodetector modules 722a, 722b, 722c, and 722d. Figure 7C shows a photodetector system having five wavelength separators 731a, 731b, 731c, 731d, and 731e. Each wavelength separator is optically coupled to photodetector modules 732a, 732b, 732c, 732d, and 732e. Figure 7D shows a photodetector system having six wavelength separators 741a, 741b, 741c, 741d, 741e, and 741f. Each wavelength separator is optically coupled to a photodetector module 742a, 742b, 742c, 742d, 742e, and 742f. Figure 7E shows a three-dimensional view of the photodetector system shown in Figure 7D.

[0066] A system for measuring the light emitted by a sample. Aspects of the present disclosure also include systems for measuring light from a sample (e.g., in a flow stream within a flow cytometer). In certain embodiments, the system includes a light source and a clustered wavelength division photodetector system, the clustered wavelength division photodetector system comprising three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, and one or more photodetector modules optically communicating with each wavelength separator, each having a plurality of photodetectors and optical components for transmitting light having a predetermined subspectral range to the photodetectors. In some embodiments, the system is a flow cytometer. In some examples, the photodetector system is indestructibly integrated into the flow cytometer. In certain embodiments, the photodetector system is optically communicating with the sample source (e.g., a flow stream within a flow cytometer) through a focusing system (e.g., an optical fiber or a free-space optical relay system).

[0067] The system for measuring light from a sample includes a light source. In embodiments, the light source may be any suitable broadband or narrowband light source. Depending on the components in the sample (e.g., cells, beads, non-cellular particles, etc.), the light source may be configured to emit light with wavelengths varying in the ranges of 200 nm to 1500 nm, e.g., 250 nm to 1250 nm, e.g., 300 nm to 1000 nm, e.g., 350 nm to 900 nm, and 400 nm to 800 nm. For example, the light source may include a broadband light source that emits light with wavelengths in the range of 200 nm to 900 nm. In other examples, the light source may include a narrowband light source that emits wavelengths in the range of 200 nm to 900 nm. For example, the light source may be a narrowband LED (1 nm to 25 nm) that emits light with wavelengths in the range of 200 nm to 900 nm.

[0068] In some embodiments, the light source is a laser. The laser in question may include pulsed lasers or continuous-wave lasers. For example, the lasers include gas lasers, such as helium-neon lasers, argon lasers, krypton lasers, xenon lasers, nitrogen lasers, CO2 lasers, CO lasers, argon-fluorine (ArF) excimer lasers, krypton-fluorine (KrF) excimer lasers, xenon-chlorine (XeCl) excimer lasers, or xenon-fluorine (XeF) excimer lasers or combinations thereof, dye lasers, such as stilbene, coumarin, or rhodamine lasers, and metal-vapor lasers, such as helium-cadmium (HeCd) lasers, helium-mercury (HeHg) lasers, helium-selenium (HeSe) lasers, and helium-silver (HeAg) lasers. This may include strontium lasers, neon-copper (NeCu) lasers, copper lasers, or gold lasers and combinations thereof, as well as solid-state lasers such as ruby ​​lasers, Nd:YAG lasers, NdCrYAG lasers, Er:YAG lasers, Nd:YLF lasers, Nd:YVO4 lasers, Nd:YCa4O(BO3)3 lasers, Nd:YCOB lasers, titanium-sapphire lasers, thallium-YAG lasers, ytterivium-YAG lasers, Y2O3 lasers, or cerium-doped lasers and combinations thereof, and implementation forms of doubling or tripling the frequency of semiconductor diode lasers, optically pumped semiconductor lasers (OPSLs), or any of the above-mentioned lasers.

[0069] In other embodiments, the light source is a non-laser light source such as a lamp including but not limited to a halogen lamp, a deuterium arc lamp, or a xenon arc lamp; a light-emitting diode such as a broadband LED having a continuous spectrum; a superluminescent light-emitting diode; a semiconductor light-emitting diode; a broad-spectrum LED white light source; or an integrated multi-LED. In some examples, the non-laser light source is a stabilized fiber-coupled broadband light source, a white light source, or any combination thereof, among other light sources.

[0070] In certain embodiments, the light source is a light beam generator configured to produce two or more beams of frequency-shifted light. In some examples, the light beam generator includes a laser and a high-frequency generator configured to apply a high-frequency drive signal to an acousto-optical device to generate two or more angularly deflected laser beams. In these embodiments, the laser may be a pulsed laser or a continuous-wave laser. For example, the laser in the light beam generator in question may be a gas laser, e.g., a helium-neon laser, argon laser, krypton laser, xenon laser, nitrogen laser, CO2 laser, CO laser, argon-fluorine (ArF) excimer laser, krypton-fluorine (KrF) excimer laser, xenon-chlorine (XeCl) excimer laser or xenon-fluorine (XeF) excimer laser or a combination thereof, a dye laser, e.g., stilbene, coumarin or rhodamine laser, or a metal-vapor laser, e.g., a helium-cadmium (HeCd) laser, a helium-mercury (HeH) laser. g) Lasers, helium-selenium (HeSe) lasers, helium-silver (HeAg) lasers, strontium lasers, neon-copper (NeCu) lasers, copper lasers, or gold lasers and combinations thereof, and solid-state lasers, such as ruby ​​lasers, Nd:YAG lasers, NdCrYAG lasers, Er:YAG lasers, Nd:YLF lasers, Nd:YVO4 lasers, Nd:YCa4O(BO3)3 lasers, Nd:YCOB lasers, titanium-sapphire lasers, thallium-YAG lasers, ytterivium-YAG lasers, Y2O3 lasers, or cerium-doped lasers and combinations thereof.

[0071] The acousto-optic device can be any suitable acousto-optic protocol configured to frequency-shift laser light using applied sound waves. In certain embodiments, the acousto-optic device is an acousto-optic deflector. The acousto-optic device in this system is configured to generate an angularly deflected laser beam from light from a laser and an applied radio frequency drive signal. The radio frequency drive signal can be applied to the acousto-optic device using any suitable radio frequency drive signal source, such as a direct digital synthesizer (DDS), an arbitrary waveform generator (AWG), or an electrical pulse generator.

[0072] In the embodiment, the controller is configured to apply radio frequency drive signals to an acoustic-optical device to generate a desired number of angularly deflected laser beams within the output laser beam, and includes being configured to apply, for example, three or more high-frequency drive signals, for example, four or more high-frequency drive signals, for example, five or more high-frequency drive signals, for example, six or more high-frequency drive signals, for example, seven or more high-frequency drive signals, for example, eight or more high-frequency drive signals, for example, nine or more high-frequency drive signals, for example, ten or more high-frequency drive signals, for example, fifteen or more high-frequency drive signals, for example, twenty-five or more high-frequency drive signals, for example, fifty or more high-frequency drive signals, and being configured to apply one hundred or more high-frequency drive signals.

[0073] In some examples, to generate an intensity profile of an angularly deflected laser beam in the output laser beam, the controller is configured to apply a high-frequency drive signal with an amplitude ranging from 5V to approximately 25V, such as approximately 0.001V to approximately 500V, for example approximately 0.005V to approximately 400V, for example approximately 0.01V to approximately 300V, for example approximately 0.05V to approximately 200V, for example approximately 0.1V to approximately 100V, for example approximately 0.5V to approximately 75V, for example approximately 1V to 50V, for example approximately 2V to 40V, for example approximately 3V to approximately 30V, etc. Each applied high-frequency drive signal has a frequency range of approximately 0.001 MHz to approximately 500 MHz, for example approximately 0.005 MHz to approximately 400 MHz, for example approximately 0.01 MHz to approximately 300 MHz, for example approximately 0.05 MHz to approximately 200 MHz, for example approximately 0.1 MHz to approximately 100 MHz, for example approximately 0.5 MHz to approximately 90 MHz, for example approximately 1 MHz to approximately 75 MHz, for example approximately 2 MHz to approximately 70 MHz, for example approximately 3 MHz to approximately 65 MHz, for example approximately 4 MHz to approximately 60 MHz, and approximately 5 MHz to approximately 50 MHz.

[0074] In certain embodiments, the controller has a processor having memory operably coupled to the processor, the memory containing instructions stored thereon, which, when executed by the processor, cause the processor to generate an output laser beam with an angle-deflected laser beam having a desired intensity profile. For example, the memory may contain instructions to generate two or more angle-deflected laser beams having the same intensity, e.g., three or more, e.g., four or more, e.g., five or more, e.g., ten or more, e.g., 25 or more, e.g., 50 or more, and the memory may contain instructions to generate 100 or more angle-deflected laser beams having the same intensity. In other embodiments, the instructions may contain instructions to generate two or more angle-deflected laser beams having different intensities, e.g., three or more, e.g., four or more, e.g., five or more, e.g., ten or more, e.g., 25 or more, e.g., 50 or more, and the memory may contain instructions to generate 100 or more angle-deflected laser beams having different intensities.

[0075] In certain embodiments, the controller has a processor having memory operably coupled to the processor, the memory containing instructions stored thereon, which, when executed by the processor, cause the processor to generate an output laser beam having an intensity that increases from the edge to the center of the output laser beam along the horizontal axis. In these examples, the intensity of the angularly deflected laser beam at the center of the output beam may be in the range of 0.1% to about 99% of the intensity of the angularly deflected laser beam at the edge of the output laser beam along the horizontal axis, for example, 0.5% to about 95%, 1% to about 90%, about 2% to about 85%, about 3% to about 80%, about 4% to about 75%, about 5% to about 70%, about 6% to about 65%, about 7% to about 60%, about 8% to about 55%, and may include about 10% to about 50% of the intensity of the angularly deflected laser beam at the edge of the output laser beam along the horizontal axis. In other embodiments, the controller has a processor having memory operably coupled to the processor, the memory containing instructions stored thereon, and when an instruction is executed by the processor, the processor outputs an output laser beam having an intensity that increases from the edge to the center of the output laser beam along the horizontal axis. In these examples, the intensity of the angularly deflected laser beam at the edge of the output beam may be in the range of 0.1% to about 99% of the intensity of the angularly deflected laser beam at the center of the output laser beam along the horizontal axis, for example, 0.5% to about 95%, 1% to about 90%, about 2% to about 85%, about 3% to about 80%, about 4% to about 75%, about 5% to about 70%, about 6% to about 65%, about 7% to about 60%, and about 8% to about 55%, and may include about 10% to about 50% of the intensity of the angularly deflected laser beam at the center of the output laser beam along the horizontal axis. In yet another embodiment, the controller has a processor having memory operably coupled to the processor, the memory containing instructions stored thereon, which, when executed by the processor, cause the processor to generate an output laser beam having an intensity profile with a Gaussian distribution along the horizontal axis. In yet another embodiment, the controller has a processor having memory operably coupled to the processor, the memory containing instructions stored thereon, which, when executed by the processor, cause the processor to generate an output laser beam having a top-hat intensity profile along the horizontal axis.

[0076] In some embodiments, the target light beam generator may be configured to generate an angle-deflected laser beam within a spatially separated output laser beam. Depending on the applied high-frequency drive signal and the desired irradiation profile of the output laser beam, the angle-deflected laser beams may be separated by 0.001 μm or more, e.g., 0.005 μm or more, e.g., 0.01 μm or more, e.g., 0.05 μm or more, e.g., 0.1 μm or more, e.g., 0.5 μm or more, e.g., 1 μm or more, e.g., 5 μm or more, e.g., 10 μm or more, e.g., 100 μm or more, e.g., 500 μm or more, e.g., 10 and even 5000 μm or more. In some embodiments, the system is configured to generate an angle-deflected laser beam within the output laser beam that overlaps with an adjacent angle-deflected laser beam along the horizontal axis of the output laser beam. The overlap between adjacent angle-deflected laser beams (such as beam spot overlap) may be an overlap of 0.001 μm or more, for example, an overlap of 0.005 μm or more, for example, an overlap of 0.01 μm or more, for example, an overlap of 0.05 μm or more, for example, an overlap of 0.1 μm or more, for example, an overlap of 0.5 μm or more, for example, an overlap of 1 μm or more, for example, an overlap of 5 μm or more, for example, an overlap of 10 μm or more, and includes overlaps of 100 μm or more.

[0077] In certain examples, a light beam generator configured to produce two or more beams of frequency-shifted light includes laser excitation modules such as those described in U.S. Patent Nos. 9,423,353, 9,784,661 and 10,006,852, and U.S. Patent Publications 2017 / 0133857 and 2017 / 0350803, the disclosures of which are incorporated herein by reference.

[0078] The light source may be positioned at any suitable distance from the sample (e.g., the flow stream in the flow cytometer), for example, at a distance of 0.001 mm or more from the flow stream, for example, at a distance of 0.005 mm or more, for example, at a distance of 0.01 mm or more, for example, at a distance of 0.05 mm or more, for example, at a distance of 0.1 mm or more, for example, at a distance of 0.5 mm or more, for example, at a distance of 1 mm or more, for example, at a distance of 5 mm or more, for example, at a distance of 10 mm or more, for example, at a distance of 25 mm or more, and at a distance of 100 mm. In addition, the light source irradiates the sample at any suitable angle (for example, with respect to the vertical axis of the flow stream), such as an angle in the range of 10° to 90°, 15° to 85°, 20° to 80°, 25° to 75° (including 30° to 60°), for example, an angle of 90°.

[0079] The light source may be configured to irradiate the sample continuously or at discrete intervals. In some examples, the system includes a light source configured to irradiate the sample continuously, such as using a continuous-wave laser that continuously irradiates a flow stream at the inspection point in a flow cytometer. In other examples, the system in question includes a light source configured to irradiate the sample at discrete intervals, such as every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds (including every 1000 milliseconds), or any other interval. When the light source is configured to irradiate the sample at discrete intervals, the system may include one or more additional components to provide intermittent irradiation of the sample by the light source. For example, the system in these embodiments may include one or more laser beam choppers, manually controlled or computer-controlled beam stops for blocking the sample and exposing it to the light source.

[0080] In embodiments, light emitted by a sample is transmitted to a photodetector system (as described above) of the present invention having two or more photodetector arrays. As described above, the photodetectors in the photodetector of the present invention may include, but are not limited to, active pixel sensors (APS), avalanche photodiodes, image sensors, charge-coupled devices (CCDs), sensitized charge-coupled devices (ICCDs), light-emitting diodes, photon counters, bolometers, pyroelectric detectors, photoresistors, photocells, photodiodes, photomultiplier tubes, phototransistors, quantum dot photoconductors or photodiodes, and combinations thereof. For example, a light-gathering system for measuring light from a sample may include a photodetector array having two or more photodetectors, e.g., three or more photodetectors, e.g., four or more photodetectors, e.g., five or more photodetectors, e.g., ten or more photodetectors, e.g., twenty-five or more photodetectors, and fifty or more photodetectors. In a particular embodiment, the system includes a photodetector array having five photodetectors.

[0081] In embodiments of the present disclosure, the detector in question is configured to measure light collected at one or more wavelengths, e.g., two or more wavelengths, e.g., five or more different wavelengths, e.g., ten or more different wavelengths, e.g., 25 or more different wavelengths, e.g., fifty or more different wavelengths, e.g., 100 or more different wavelengths, e.g., 200 or more different wavelengths, e.g., 300 or more different wavelengths, and includes measuring light emitted by a sample in a flow stream at 400 or more different wavelengths.

[0082] In embodiments, the photodetector of a photodetection system is configured to measure light continuously or at discrete intervals. In some examples, the detector in question is configured to measure the collected light continuously. In other examples, the detector in question is configured to measure light at discrete intervals, such as every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds (including every 1000 milliseconds), or some other interval.

[0083] In some embodiments, a system for measuring light from a sample includes a focusing system for collecting light from a sample source (e.g., a flowstream) and directing it to a target photodetector. The focusing system may be physically coupled to the photodetector using an adhesive or the like, co-molded with the photodetector, or integrated into the photodetector. In certain embodiments, the focusing system and the photodetector are integrated into a single unit. In other embodiments, the focusing system is coupled to the photodetector using connectors such as hook-and-loop fasteners, magnets, latches, notches, countersunk holes, counterbores, grooves, pins, tethers, hinges, Velcro, non-permanent adhesives, or a combination thereof.

[0084] In other embodiments, the light detection system and the light focusing system are in optical communication but not in physical contact. For example, the light focusing system may be positioned at a distance of 0.001 mm or more from the light detection system, e.g., 0.005 mm or more, e.g., 0.01 mm or more, e.g., 0.05 mm or more, e.g., 0.1 mm or more, e.g., 0.5 mm or more, e.g., 1 mm or more, e.g., 10 mm or more, e.g., 25 mm or more, e.g., 50 mm or more, and 100 mm or more from the light detection system.

[0085] In some embodiments, the focusing system includes optical fibers. For example, in some cases, the focusing system may be an optical fiber relay bundle through which light is transmitted to a photodetector. In other embodiments, the focusing system is a free-space optical relay system. For example, a free-space optical relay system may include a housing having a proximal end and a distal end, the proximal end being coupled to a photodetector. A free-space relay system may include any combination of different optical components, such as one or more lenses, mirrors, slits, pinholes, wavelength separators, or combinations thereof.

[0086] In certain embodiments, the system is a flow cytometry system that uses the above-described photodetection system for detecting light emitted by a sample in a flow stream. Suitable flow cytometry systems include: Ormerod (ed.), Flow Cytometry: A Practical Approach, Oxford Univ. Press (1997); Jaroszeski et al. (eds.), Flow Cytometry Protocols, Methods in Molecular Biology No. 91, Humana Press (1997); Practical Flow Cytometry, 3rd ed., Wiley-Liss (1995); Virgo, et al. (2012) Ann Clin Biochem. Jan; 49 (pt 1): 17-28; Linden, et al., Semin Throm Hemost. 2004 Oct; 30 (5): 502-11; Alison, et al. J Pathol, 2010 Dec; 222 (4): 335-344; and Herbig, et al. (2007) Crit Rev Ther Drug Carrier Examples include, but are not limited to, those described in Syst.24(3):203-255;, and these disclosures are incorporated herein by reference.In specific cases, the flow cytometry systems covered include BD Biosciences FACSCanto® II flow cytometer, BD Accuri® flow cytometer, BD Biosciences FACSCelesta® flow cytometer, BD Biosciences FACSLyric® flow cytometer, BD Biosciences FACSVerse® flow cytometer, BD Biosciences FACSymphony® flow cytometer, BD Biosciences LSRFortessa® flow cytometer, BD Biosciences LSRFortess® X-20 flow cytometer, and BD Biosciences FACSCalibur® flow cytometer, BD Biosciences FACSCount® cell sorter, BD Biosciences FACSLyric® cell sorter, and BD Biosciences Via® cell sorter, BD Biosciences Influx® cell sorter, BD Biosciences Jazz® cell sorter, BD Biosciences Aria® cell sorter, and BD Examples include Biosciences FACSMelody® cell sorting machine.

[0087] In some embodiments, the subject particle sorting system is specified in U.S. Patent Nos. 10,006,852, 9,952,076, 9,933,341, 9,784,661, 9,726,527, 9,453,789, 9,200,334, 9,097,640, 9,095,494, 9,092,034, 8,975,595, 8,753,573, 8,233,146, 8,140,300, and 7 Flow cytometry systems such as those described in Nos. 544,326, 7,201,875, 7,129,505, 6,821,740, 6,813,017, 6,809,804, 6,372,506, 5,700,692, 5,643,796, 5,627,040, 5,620,842, and 5,602,039 are incorporated herein by reference in their entirety.

[0088] In specific examples, the subject system is a flow cytometry system configured to image particles in a flow stream by fluorescence imaging using radio frequency tagged emission (FIRE), such as those described in Diebold, et al. Nature Photonics Vol. 7(10); 806-810 (2013), and in U.S. Patents No. 9,423,353, No. 9,784,661, and No. 10,006,852, and U.S. Patent Publication Nos. 2017 / 0133857 and 2017 / 0350803, the disclosures of which are incorporated herein by reference.

[0089] Method for measuring light collected from an irradiated sample Aspects of the present disclosure also include methods for measuring light from a sample (for example, in a flow stream in a flow cytometer). When carrying out the methods according to the embodiments, as described above, the sample is irradiated with a light source, and the light from the sample is detected in a photodetection system having three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, and one or more photodetection modules optically communicating with each wavelength separator, each having a plurality of photodetectors and optical components that transmit light having a predetermined subspectral range to the photodetectors. In some embodiments, the sample is a biological sample. The term “biological sample” is used in its conventional sense to refer to a whole organism, plant, fungus, or, in specific examples, a subset of animal tissues, cells, or components that may be found in blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, bronchoalveolar lavage, amniotic fluid, amniotic umbilical cord blood, urine, vaginal fluid, and semen. Therefore, “biological sample” refers to both a natural organism or a subset of its tissues, as well as homogenates, lysates, or extracts prepared from a subset of an organism or its tissues, including, but not limited to, plasma, serum, cerebrospinal fluid, lymph, skin sections, respiratory tract, gastrointestinal tract, cardiovascular and urinary tract, tears, saliva, milk, blood cells, tumors, and organs. A biological sample can be any type of biological tissue, including both healthy tissue and diseased tissue (e.g., cancerous, malignant, necrotic, etc.). In certain embodiments, a biological sample is a liquid sample such as blood or its derivatives, e.g., plasma, tears, urine, semen, and in some examples, the sample is a blood sample including whole blood, such as blood obtained from a venipuncture or fingerstick (blood may or may not be combined with any reagents such as preservatives and anticoagulants before the assay).

[0090] In certain embodiments, the source of the sample is “mammal” or “mammalian,” terms widely used to refer to organisms within the class Mammalia, including Carnivora (e.g., dogs and cats), Rodentia (e.g., mice, guinea pigs, and rats), and Primates (e.g., humans, chimpanzees, and monkeys). In some examples, the subject is human. The method may be applied to samples obtained from human subjects of both sexes at any stage of development (i.e., neonates, infants, adolescents, and adults), and in certain embodiments, the human subject is adolescent, adolescent, or adult. While the present invention may be applied to samples from human subjects, it should be understood that it may also be applied to samples from other animal subjects (i.e., “non-human subjects”), such as birds, mice, rats, dogs, cats, livestock, and horses, but is not limited to the above.

[0091] In carrying out the method of the present invention, the sample (e.g., in the flow stream of a flow cytometer) is irradiated with light from a light source. In some embodiments, the light source is a broadband light source that emits light having a wide range of wavelengths, for example, from 50 nm or more, 100 nm or more, 150 nm or more, 200 nm or more, 250 nm or more, 300 nm or more, 350 nm or more, 400 nm or more, and 500 nm or more. For example, one suitable broadband light source emits light having wavelengths from 200 nm to 1500 nm. Another embodiment of a suitable broadband light source includes a light source that emits light having wavelengths from 400 nm to 1000 nm. If the method involves illumination with a broadband light source, the broadband light source protocol in question may include, but is not limited to, halogen lamps, deuterium arc lamps, xenon arc lamps, stabilized fiber-coupled broadband light sources, broadband LEDs with continuous spectra, ultra-high brightness light-emitting diodes, semiconductor light-emitting diodes, broad-spectrum LED white light sources, multi-LED integrated white light sources, or any combination thereof.

[0092] In other embodiments, the method includes irradiating with a narrowband light source that emits a specific wavelength or a narrow wavelength range, such as a light source that emits light in a narrow wavelength range such as 50 nm or less, 40 nm or less, 30 nm or less, 25 nm or less, 20 nm or less, 15 nm or less, 10 nm or less, 5 nm or less, or 2 nm or less, and includes a light source that emits light of a specific wavelength (i.e., monochromatic light). If the method includes irradiating with a narrowband light source, the narrowband light source protocol in question may include, but is not limited to, a narrow-wavelength LED, a laser diode, or a broadband light source coupled to one or more optical bandpass filters, diffraction gratings, monochromators, or any combination thereof.

[0093] In certain embodiments, the method includes irradiating a sample with one or more lasers. As described above, the type and number of lasers vary depending on the sample and the desired light to be collected, and may be gas lasers such as helium-neon lasers, argon lasers, krypton lasers, xenon lasers, nitrogen lasers, CO2 lasers, CO lasers, argon-fluorine (ArF) excimer lasers, krypton-fluorine (KrF) excimer lasers, xenon-chlorine (XeCl) excimer lasers, or xenon-fluorine (XeF) excimer lasers, or combinations thereof. In other examples, the method includes irradiating a flow stream with dye lasers such as stilbene, coumarin, or rhodamine lasers. In yet another example, the method includes irradiating the flowstream with a metal-vapor laser such as a helium-cadmium (HeCd) laser, a helium-mercury (HeHg) laser, a helium-selenium (HeSe) laser, a helium-silver (HeAg) laser, a strontium laser, a neon-copper (NeCu) laser, a copper laser, or a gold laser, or a combination thereof. In yet another example, the method includes irradiating the flowstream with a solid-state laser, such as a ruby ​​laser, Nd:YAG laser, NdCrYAG laser, Er:YAG laser, Nd:YLF laser, Nd:YVO4 laser, Nd:YCa4O(BO3)3 laser, Nd:YCOB laser, a titanium-sapphire laser, a thallium-YAG laser, a ytterivium-YAG laser, a Y2O3 laser, or a cesium-doped laser, or a combination thereof.

[0094] The sample may be irradiated with one or more of the above-mentioned light sources, for example, two or more light sources, for example, three or more light sources, for example, four or more light sources, for example, five or more light sources, for example, ten or more light sources. The light sources may include any combination of light source types. For example, in some embodiments, the method includes irradiating the sample in the flow stream with an array of lasers, such as an array having one or more gas lasers, one or more dye lasers, and one or more solid-state lasers.

[0095] The sample may be irradiated with wavelengths in the range of 200 nm to 1500 nm, for example 250 nm to 1250 nm, for example 300 nm to 1000 nm, for example 350 nm to 900 nm, for example 400 nm to 800 nm. For example, if the light source is a broadband light source, the sample may be irradiated with wavelengths in the range of 200 nm to 900 nm. In other examples where the light source includes multiple narrowband light sources, the sample may be irradiated with specific wavelengths in the range of 200 nm to 900 nm. For example, the light source may be multiple narrowband LEDs (1 nm to 25 nm), each narrowband LED independently emitting light having a wavelength range of 200 nm to 900 nm. In other embodiments, the narrowband light source includes one or more lasers (such as a laser array), and the sample is irradiated with specific wavelengths in the range of 200 nm to 700 nm using a laser array, such as a gas laser, excimer laser, dye laser, metal vapor laser, and solid-state laser as described above.

[0096] When two or more light sources are used, the sample may be irradiated simultaneously, sequentially, or in combination thereof using the light sources. For example, the sample may be irradiated simultaneously by each light source. In other embodiments, the flowstream is irradiated sequentially by each light source. When the sample is irradiated sequentially using two or more light sources, the time each light source irradiates the sample may be independently 0.001 microseconds or more, e.g., 0.01 microseconds or more, e.g., 0.1 microseconds or more, e.g., 1 microsecond or more, e.g., 5 microseconds or more, e.g., 10 microseconds or more, e.g., 30 microseconds or more, and including 60 microseconds or more. For example, the method may include irradiating the sample with a light source (e.g., a laser) for a duration in the range of 0.001 microseconds to 100 microseconds, e.g., 0.01 microseconds to 75 microseconds, e.g., 0.1 microseconds to 50 microseconds, e.g., 1 microsecond to 25 microseconds (including 5 microseconds to 10 microseconds). In embodiments in which the sample is irradiated sequentially by two or more light sources, the durations of irradiation of the sample by each light source may be the same or different.

[0097] The interval between irradiations from each light source may also vary as needed and be independently separated by delays of 0.001 microseconds or more, e.g., 0.01 microseconds or more, e.g., 0.1 microseconds or more, e.g., 1 microsecond or more, e.g., 5 microseconds or more, e.g., 10 microseconds or more, e.g., 15 microseconds or more, e.g., 30 microseconds or more, and 60 microseconds or more. For example, the interval between irradiations from each light source may be in the range of 0.001 microseconds to 60 microseconds, e.g., 0.01 microseconds to 50 microseconds, e.g., 0.1 microseconds to 35 microseconds, e.g., 1 microsecond to 25 microseconds, and 5 microseconds to 10 microseconds. In a particular embodiment, the interval between irradiations from each light source is 10 microseconds. In embodiments in which the sample is sequentially irradiated by more than two (i.e., three or more) light sources, the delays between irradiations from each light source may be the same or different.

[0098] The sample can be irradiated continuously or at discontinuous intervals. In some examples, the method involves continuously irradiating the sample with a light source. In other examples, the sample is irradiated with a light source at discrete intervals, such as every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds (including every 1000 milliseconds), or some other intervals.

[0099] Depending on the light source, the sample may be irradiated from a distance that varies, including, for example, 0.01 mm or more, 0.05 mm or more, 0.1 mm or more, 0.5 mm or more, 1 mm or more, 2.5 mm or more, 5 mm or more, 10 mm or more, 15 mm or more, 25 mm or more, and 50 mm or more. The angle or irradiation may also vary within the range of 10° to 90°, for example 15° to 85°, for example 20° to 80°, for example 25° to 75°, including 30° to 60°, and for example, an angle of 90°.

[0100] As described above, in the embodiment, light from an irradiated sample is transmitted to a photodetection system described herein and measured by one or more photodetectors. When carrying out this method, light from the sample is transmitted to three or more wavelength separators, each configured to transmit light having a predetermined spectral range. Light of a spectral range from each wavelength separator is transmitted to one or more photodetection modules having optical components configured to transmit light having a predetermined subspectral range to a photodetector.

[0101] Light can be measured continuously or at discrete intervals using a photodetector system. In some examples, the method involves continuously measuring the light. In other examples, the light is measured at discrete intervals, such as every 0.001 milliseconds, every 0.01 milliseconds, every 0.1 milliseconds, every 1 millisecond, every 10 milliseconds, every 100 milliseconds (including every 1000 milliseconds), or at some other interval.

[0102] The collected light may be measured one or more times during the method, for example, two or more times, for example, three or more times, for example, five or more times, for example, ten or more times. In certain embodiments, light propagation is measured two or more times, and the data in a particular example is averaged.

[0103] In some embodiments, the method includes aligning the light before detecting it using the photodetector system of the present invention. For example, light from a sample source may pass through one or more lenses, mirrors, pinholes, slits, gratings, optical refractors, and any combination thereof. In some examples, the collected light passes through one or more focusing lenses, for example, to reduce the profile of the light directed to the photodetector system or focusing system as described above. In other examples, the light emitted from the sample passes through one or more collimators to reduce the divergence of the light beam transmitted to the photodetector system.

[0104] kit Aspects of the present invention further include a kit comprising three or more wavelength separators, a plurality of photodetectors, and one or more optical components (e.g., dichroic mirrors, beam splitters, collimating lenses, etc.). In some embodiments, the kit includes a substrate for co-mounting the wavelength separators with the optical components and photodetectors. In certain embodiments, the kit includes one or more fasteners for assembling the components of the subject photodetector system together. The kit may also include a focusing component such as an optical fiber (e.g., an optical fiber relay bundle) or components for a free-space relay system. In some examples, the kit further includes one or more photodetectors, such as a photomultiplier tube (e.g., a metal-packaged photomultiplier tube).

[0105] In some embodiments, the kit comprises two or more components of the photodetection system disclosed herein, for example, three or more (including five or more). In some cases, the kit may comprise one or more assay components (e.g., labeling reagents, buffers, etc., as described above). In some examples, the kit may further comprise, if necessary, a sample collection device, for example, a lance or needle configured to puncture the skin to obtain a whole blood sample, a pipette, etc.

[0106] The various assay components of the kit may be located in separate containers, or some or all of them may be pre-assembled. For example, in some cases, one or more components of the kit may be located in a sealed pouch, such as a sterile foil pouch or envelope.

[0107] In addition to the components described above, the subject kit may further include instructions for carrying out the subject method (in a particular embodiment). These instructions may be present in the subject kit in various forms, and one or more of these forms may be present in the kit. One form in which these instructions may be present is, for example, as printed information in the kit packaging, such as on a suitable medium or substrate such as one or more sheets of paper on which the information is printed, in the kit packaging, or in accompanying documents. Yet another form of these instructions may be a computer-readable medium on which the information is recorded, such as a diskette, compact disc (CD), or portable flash drive. Yet another form in which these instructions may be present is a website address that can be used via the internet to access the information at a remote site.

[0108] Utility The photodetection system of the present invention is used when it is desirable to characterize a sample by its optical properties, particularly when low levels of light are collected. In some embodiments, the systems and methods described herein find applications in flow cytometry characterization of biological samples labeled with fluorescent tags. In other embodiments, the systems and methods find applications in transmitted or scattered light spectroscopy. In addition, the systems and methods of the subject matter find applications in increasing the signal obtained from light collected from a sample (e.g., in a flow stream). In certain examples, the disclosure is used to improve the measurement of light collected from a sample irradiated in a flow stream within a flow cytometer. Embodiments of the disclosure are used when it is desirable to enhance the effectiveness of luminescence measurements in flow cytometry, such as in research and high-throughput laboratory testing. The disclosure is also used when it is desirable to provide a flow cytometer with improved cell sorting accuracy, enhanced particle collection, reduced energy consumption, particle charging efficiency, more accurate particle charging, and enhanced particle deflection during cell sorting.

[0109] This disclosure also finds applications in applications where cells prepared from biological samples may be desired for research, laboratory testing, or therapeutic use. In some embodiments, the methods and devices may facilitate the acquisition of individual cells prepared from target fluid or tissue biological samples. For example, the methods and systems of the subject facilitate the acquisition of cells from fluid or tissue samples used as specimens for research or diagnosis of diseases such as cancer. Similarly, the methods and systems of the subject facilitate the acquisition of cells from fluid or tissue samples used in therapeutic applications. The methods and devices of the disclosure enable the separation and collection of cells from biological samples (e.g., organs, tissues, tissue fragments, body fluids) with improved efficiency and lower cost compared to conventional flow cytometry systems.

[0110] Notwithstanding the attached claims, this disclosure is also defined by the following annotation: 1. A light detection system, Three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, One or more photodetector modules that are optically connected to each wavelength separator. Each photodetector module includes, Multiple photodetectors, An optical component configured to transmit light having a predetermined subspectral range to a photodetector, A light detection system, including a light detection system. 2. The photodetection system as described in Appendix 1, wherein the wavelength separators are configured to transmit light between them. 3. Each wavelength separator is a photodetection system as described in Appendix 1 or 2, including a dichroic mirror. 4. The wavelength separator is positioned along a single plane in the photodetection system described in any one of the appendices 1 to 3. 5. A photodetection system as described in any one of the appendices 1 to 3, wherein the wavelength separator is positioned along two or more planes.

[0111] 6. The photodetection system as described in Appendix 5, wherein the wavelength separator is positioned along two or more parallel planes. 7. The photodetector described in Appendix 6, wherein the wavelength separator has a polygonal configuration in the photodetector. 8. The wavelength separator is a photodetection system as described in Appendix 7, having a pentagonal or hexagonal configuration. 9. A photodetection system as described in any one of the appendices 1 to 8, wherein the wavelength separator is configured to allow light having wavelengths in the range of 200 nm to 1200 nm to pass through. 10. A photodetection system as described in any one of the appendices 1 to 9, wherein each wavelength separator is configured to allow light having a spectral range of 75 nm to 150 nm to pass through.

[0112] 11. The photodetection system as described in Appendix 10, wherein each wavelength separator is configured to allow light having a spectral range of 100 nm to pass through. 12. A photodetection system as described in any one of the appendices 1 to 11, wherein the wavelength separator is configured to allow light having wavelengths in the range of 360 nm to 960 nm to pass through. 13. Wavelength separators are, A first wavelength separator configured to allow light having wavelengths in the range of 360 nm to 480 nm to pass through, A second wavelength separator configured to allow light having wavelengths in the range of 480 nm to 600 nm to pass through, A third wavelength separator configured to allow light having wavelengths in the range of 600 nm to 720 nm to pass through, A fourth wavelength separator configured to allow light having wavelengths in the range of 720 nm to 840 nm to pass through, A fifth wavelength separator configured to allow light having wavelengths in the range of 840 nm to 960 nm to pass through, The photodetection system described in Appendix 12, including the one described in Appendix 12. 14. The first wavelength separator is configured to transmit light having wavelengths in the range of 480 nm to 960 nm to the second wavelength separator. The second wavelength separator is configured to transmit light having wavelengths in the range of 600 nm to 960 nm to the third wavelength separator. The third wavelength separator is configured to transmit light having wavelengths in the range of 720 nm to 960 nm to the fourth wavelength separator. The photodetection system as described in Appendix 13, wherein the fourth wavelength separator is configured to transmit light having wavelengths in the range of 840 nm to 960 nm to the fifth wavelength separator. 15. A photodetection system according to any one of the appendices 1 to 14, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 50 nm to pass through to the photodetector.

[0113] 16. The photodetection system as described in Appendix 15, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 25 nm to pass through to the photodetector. 17. The photodetection system as described in Appendix 15, wherein each optical component is configured to allow light having a subspectral range of 20 nm to pass through to the photodetector. 18. Each optical component is a light detection system as described in any one of the appendices 1 to 17, including a dichroic mirror. 19. A photodetection system as described in any one of Appendices 1 to 18, wherein the optical components are positioned within the photodetection module along a single plane. 20. An optical detection system according to any one of the appendices 1 to 18, wherein the optical components are positioned within the optical detection module along two or more planes.

[0114] 21. The photodetection system as described in Appendix 20, wherein the optical components are positioned within the photodetection module along two or more parallel planes. 22. The optical component has a polygonal configuration in the light detection module, as described in Appendix 20. 23. The optical component of the photodetection system as described in Appendix 22, wherein the photodetection module has a hexagonal, heptagonal, or octagonal configuration. 24. The photodetector is a photodetector system as described in any one of the appendices 15 to 23, comprising one or more photomultiplier tubes. 25. The photodetector is a photodetector system as described in any one of the appendices 15 to 23, comprising one or more photodiodes. 26. The photodetector is a photodetector comprising one or more avalanche photodiodes, as described in any one of the appendices 15 to 23.

[0115] 27. A light detection system, A wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, First, second, and third photodetector modules configured to receive light from each of the first, second, and third predetermined spectral ranges, respectively. The first, second, and third photodetector modules each include, Multiple photodetectors, An optical component configured to transmit light having a predetermined subspectral range to a photodetector, A light detection system, including a light detection system. 28. The wavelength separator is a prism, as described in Appendix 27 of the photodetection system. 29. The wavelength separator is a diffraction grating, as described in Appendix 27 of the photodetection system. 30. The photodetection system according to any one of the appendices 27 to 29, wherein the first, second, and third predetermined spectral ranges each have a spectral range of 75 nm to 150 nm. 31. The photodetection system according to any one of the appendices 27 to 29, wherein the first, second, and third predetermined spectral ranges each have a spectral range of 100 nm.

[0116] 32. A photodetection system according to any one of the appendices 27 to 31, wherein the wavelength separator is configured to generate light of a fourth predetermined spectral range from a light source. 33. The photodetection system according to Appendix 32, comprising a fourth photodetector module configured to receive light in a fourth predetermined spectral range. 34. A photodetection system according to any one of the appendices 27 to 31, wherein the wavelength separator is configured to generate light from a light source in four or more predetermined spectral ranges. 35. The photodetection system according to Appendix 34, further comprising a photodetection module for each of four or more predetermined spectral ranges of light. 36. Wavelength separators are, Light in a first predetermined spectral range of 360nm to 480nm, Light in a second predetermined spectral range of 480nm to 600nm, Light in a third predetermined spectral range of 600nm to 720nm, Light in a fourth predetermined spectral range of 720nm to 840nm, Light in a fifth predetermined spectral range of 840nm to 960nm and A photodetection system according to appendix 34 or 35, configured to generate a

[0117] 37. A photodetection system according to any one of the appendices 27 to 36, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 50 nm to pass through to the photodetector. 38. The photodetection system as described in Appendix 37, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 25 nm to pass through to the photodetector. 39. The photodetection system as described in Appendix 38, wherein each optical component is configured to allow light having a subspectral range of 20 nm to pass through to the photodetector. 40. Each optical component is a light detection system as described in any one of the appendices 27 to 39, including a dichroic mirror. 41. An optical detection system according to any one of the appendices 27 to 40, wherein the optical components are positioned within the optical detection module along a single plane.

[0118] 42. An optical detection system according to any one of the appendices 27 to 40, wherein the optical components are positioned within the optical detection module along two or more planes. 43. The photodetection system as described in Appendix 42, wherein the optical components are positioned within the photodetection module along two or more parallel planes. 44. The optical components have a polygonal configuration in the light detection module, as per Appendix 43. The described light detection system. 45. The optical component of the photodetection system as described in Appendix 44, wherein the photodetection module has a hexagonal, heptagonal, or octagonal configuration. 46. ​​The photodetector is a photodetector comprising one or more photomultiplier tubes, as described in any one of the appendices 27 to 45.

[0119] 47. The photodetector is a photodetector system as described in any one of the appendices 27 to 45, comprising one or more photodiodes. 48. The photodetector is a photodetector system according to any one of the appendices 27 to 45, comprising one or more avalanche photodiodes.

[0120] 49. A light detection system, A wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, First, second, and third photodetector modules configured to receive light from each of the first, second, and third predetermined spectral ranges, respectively. Includes, Each of the first, second, and third photodetector modules is configured to generate light in multiple subspectral ranges. A photodetector system in which light in each subspectral range exhibits less than 20% optical loss compared to light from the light source. 50. The photodetection system described in Appendix 49, wherein light in each subspectral range exhibits an optical loss of 15% or less compared to light from the light source. 51. The photodetection system according to Appendix 49 or 50, wherein the first, second, and third photodetection modules are configured to generate light in 20 or more subspectral ranges. 52. The photodetection system according to Appendix 49 or 50, wherein the first, second, and third photodetection modules each include a plurality of dichroic mirrors and are configured to generate light in 20 or more subspectral ranges with 10 or fewer reflections. 53. The wavelength separator is a prism, as described in any one of the appendices 49-52 of the photodetection system.

[0121] 54. The wavelength separator is a diffraction grating, as described in any one of the appendices 49 to 52 of the photodetection system. 55. The photodetection system according to any one of the appendices 49 to 54, wherein the first, second, and third predetermined spectral ranges each have a spectral range of 75 nm to 150 nm. 56. The photodetection system according to any one of the appendices 49 to 54, wherein the first, second, and third predetermined spectral ranges each have a spectral range of 100 nm. 57. A photodetection system according to any one of the appendices 49 to 56, wherein the wavelength separator is configured to generate light of a fourth predetermined spectral range from a light source. 58. The photodetection system according to Appendix 57, comprising a fourth photodetector module configured to receive light in a fourth predetermined spectral range.

[0122] 59. A photodetection system according to any one of the appendices 49 to 58, wherein the wavelength separator is configured to generate light from a light source in four or more predetermined spectral ranges. The photodetection system according to Appendix 59, further comprising a photodetection module for each of four or more predetermined spectral ranges of light. 61. A photodetection system according to any one of the appendices 49 to 60, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 50 nm to pass through to the photodetector. 62. Each photodetector module generates light in a subspectral range including 5 nm to 25 nm. The photodetection system described in Appendix 37, configured to be used in this way. 63. The photodetection system as described in Appendix 62, wherein each photodetector module is configured to generate light in a subspectral range including 5 nm to 20 nm.

[0123] 64. Each photodetector module is a photodetector system as described in any one of the appendices 49 to 63, comprising multiple dichroic mirrors. 65. The photodetection system as described in Appendix 64, wherein a dichroic mirror is positioned within the photodetection module along a single plane. 66. The photodetection system as described in Appendix 64, wherein a dichroic mirror is positioned within the photodetection module along two or more planes. 67. The photodetection system as described in Appendix 66, wherein the dichroic mirror is positioned within the photodetection module along two or more parallel planes. 68. The light detection system described in Appendix 66, wherein dichroic mirrors are arranged in a polygonal configuration in each light detection module. 69. The photodetection system as described in Appendix 68, wherein the dichroic mirrors are arranged in a hexagonal, heptagonal, or octagonal configuration within the photodetection module.

[0124] 70. It is a system, Light source and Light detection system and The photodetection system includes, Three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, One or more photodetector modules that are optically connected to each wavelength separator. Each photodetector module includes, Multiple photodetectors, An optical component configured to transmit light having a predetermined subspectral range to a photodetector, A system that includes this. 71. The light source is a laser, as described in Appendix 70. 72. The system is a flow cytometer, as described in Appendix 70 or 71. 73. The system described in any one of the appendices 70 to 72, further comprising a light-gathering system for propagating light to a light-detecting system. 74. The light-gathering component is the system described in Appendix 73, including an optical fiber.

[0125] 75. The light-gathering component is an optical fiber relay bundle, as described in Appendix 74. 76. The light-gathering component is a free-space light relay system, as described in Appendix 73. 77. A wavelength separator is configured to transmit light between each other, as described in Appendix 70. 78. Each wavelength separator is a system described in any one of the appendices 70-77, including a dichroic mirror. 79. The wavelength separator is positioned along a single plane, as described in any one of the appendices 70-78.

[0126] 80. The wavelength separator is positioned along two or more planes, as described in any one of the systems described in appendices 70 to 78. 81. The system described in Appendix 80, wherein the wavelength separator is positioned along two or more parallel planes. 82. The wavelength separator is a system described in Appendix 81, having a polygonal configuration in the photodetection system. 83. The wavelength separator is the system described in Appendix 82, having a pentagonal or hexagonal configuration. 84. A wavelength separator is configured to allow light having wavelengths in the range of 200 nm to 1200 nm to pass through, as described in any one of the appendices 70 to 83.

[0127] 85. A system as described in any one of the appendices 70 to 84, wherein each wavelength separator is configured to allow light having a spectral range of 75 nm to 150 nm to pass through. 86. The system as described in Appendix 85, wherein each wavelength separator is configured to allow light having a spectral range of 100 nm to pass through. 87. A wavelength separator is configured to allow light having wavelengths in the range of 360 nm to 960 nm to pass through, as described in any one of the appendices 70 to 86. 88. Wavelength separators are, A first wavelength separator configured to allow light having wavelengths in the range of 360 nm to 480 nm to pass through, A second wavelength separator configured to allow light having wavelengths in the range of 480 nm to 600 nm to pass through, A third wavelength separator configured to allow light having wavelengths in the range of 600 nm to 720 nm to pass through, A fourth wavelength separator configured to allow light having wavelengths in the range of 720 nm to 840 nm to pass through, A fifth wavelength separator configured to allow light having wavelengths in the range of 840 nm to 960 nm to pass through, The system described in Appendix 87, including the system described therein. 89. The first wavelength separator is configured to transmit light having wavelengths in the range of 480 nm to 960 nm to the second wavelength separator. The second wavelength separator is configured to transmit light having wavelengths in the range of 600 nm to 960 nm to the third wavelength separator. The third wavelength separator is configured to transmit light having wavelengths in the range of 720 nm to 960 nm to the fourth wavelength separator. The system as described in Appendix 87, wherein the fourth wavelength separator is configured to transmit light having wavelengths in the range of 840 nm to 960 nm to the fifth wavelength separator.

[0128] 90. The system described in any one of the appendices 70 to 89, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 50 nm to pass through to a photodetector. 91. The system as described in Appendix 90, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 25 nm to pass through to a photodetector. 92. The system as described in Appendix 90, wherein each optical component is configured to allow light having a subspectral range of 20 nm to pass through to a photodetector. 93. Each optical component is a system described in any one of the appendices 70 to 92, including a dichroic mirror. 94. The system described in any one of the appendices 70 to 93, wherein the optical components are positioned within the photodetector module along a single plane.

[0129] 95. The system described in any one of the appendices 70 to 94, wherein the optical components are positioned within the photodetector module along two or more planes. 96. The system as described in Appendix 95, wherein the optical components are positioned within the photodetector module along two or more parallel planes. 97. The optical component has a polygonal configuration in the light detection module, as per Appendix 96. The system described. 98. The system as described in Appendix 97, wherein the optical components have a hexagonal, heptagonal, or octagonal configuration in the light detection module. 99. The photodetector is a system described in any one of the appendices 90 to 98, comprising one or more photomultiplier tubes.

[0130] 100. The photodetector is a system described in any one of the appendices 90 to 99, comprising one or more photodiodes. 101. The photodetector is a system described in any one of the appendices 90 to 99, comprising one or more avalanche photodiodes.

[0131] 102. A system, Light source and Light detection system and The photodetection system includes, A wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, First, second, and third photodetector modules configured to receive light from each of the first, second, and third predetermined spectral ranges, respectively. The first, second, and third photodetector modules each include, Multiple photodetectors, An optical component configured to transmit light having a predetermined subspectral range to a photodetector, A system that includes this. 103. The light source is a laser, as described in Appendix 102. 104. The system is a flow cytometer, as described in Appendix 102 or 103. 105. A system as described in any one of the appendices 102 to 104, further comprising a light-gathering system for propagating light to a light-detecting system. 106. The light-gathering component is the system described in Appendix 105, including an optical fiber.

[0132] 107. The focusing component is an optical fiber relay bundle, as described in Appendix 106. 108. The light-gathering component is a free-space light relay system, as described in Appendix 105. 109. The wavelength separator is a prism, as described in any one of the appendices 102-108. 110. The wavelength separator is a diffraction grating, as described in any one of the appendices 102 to 108. 111. The system described in any one of the appendices 102 to 110, wherein the first, second, and third predetermined spectral ranges each have a spectral range of 75 nm to 150 nm.

[0133] 112. The system as described in Appendix 111, wherein the first, second, and third predetermined spectral ranges each have a spectral range of 100 nm. 113. A wavelength separator configured to generate light of a fourth predetermined spectral range from a light source, as described in any one of the systems described in Appendix 102 to 112. 114. The photodetection system, as described in Appendix 113, includes a fourth photodetection module configured to receive light in a fourth predetermined spectral range. 115. A wavelength separator is configured to generate light from a light source in four or more predetermined spectral ranges, as described in any one of the systems described in Appendix 102 to 114. 116. The system according to Appendix 115, further comprising a photodetector module for each of four or more predetermined spectral ranges of light.

[0134] 117. Wavelength separators are, Light in a first predetermined spectral range of 360nm to 480nm, Light in a second predetermined spectral range of 480nm to 600nm, Light in a third predetermined spectral range of 600nm to 720nm, Light in a fourth predetermined spectral range of 720nm to 840nm, Light in a fifth predetermined spectral range of 840nm to 960nm and The system described in Appendix 115 or 116, configured to generate the following: 118. A system as described in any one of the appendices 102 to 117, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 50 nm to pass through a photodetector. 119. The system as described in Appendix 118, wherein each optical component is configured to allow light having a subspectral range of 5 nm to 25 nm to pass through to a photodetector. 120. The system as described in Appendix 119, wherein each optical component is configured to pass light having a subspectral range of 20 nm through a photodetector. 121. Each optical component is a system described in any one of the appendices 102 to 120, including a dichroic mirror.

[0135] 122. The system described in any one of appendices 102 to 121, wherein the optical components are positioned within the photodetector module along a single plane. 123. The system described in any one of the appendices 102 to 121, wherein the optical components are positioned within the photodetector module along two or more planes. 124. The system as described in Appendix 123, wherein the optical adjustment components are positioned within the light detection module along two or more parallel planes. 125. The system as described in Appendix 124, wherein the optical component has a polygonal configuration in the light detection module. 126. The system as described in Appendix 125, wherein the optical components have a hexagonal, heptagonal, or octagonal configuration in the light detection module.

[0136] 127. The photodetector is a system described in any one of the appendices 102 to 126, comprising one or more photomultiplier tubes. 128. The photodetector is a system described in any one of the appendices 102 to 126, comprising one or more photodiodes. 129. The photodetector is a system described in any one of the appendices 102 to 126, comprising one or more avalanche photodiodes.

[0137] 130. A system, Light source and Light detection system and The photodetection system includes, A wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, First, second, and third photodetector modules configured to receive light from each of the first, second, and third predetermined spectral ranges, respectively. Includes, Each of the first, second, and third photodetector modules is configured to generate light in multiple subspectral ranges. A system in which light in each subspectral range exhibits less than 20% optical loss compared to light from the light source. 131. The system described in Appendix 130, wherein the light in each subspectral range exhibits no more than 15% optical loss compared to the light from the light source. 132. The system according to Appendix 130 or 131, wherein the first, second, and third photodetector modules are configured to generate light in 20 or more subspectral ranges. 133. The system according to Appendix 130 or 131, wherein the first, second, and third photodetector modules each include a plurality of dichroic mirrors and are configured to generate light in 20 or more subspectral ranges with 10 or fewer reflections. 134. The light source is a laser, as described in Appendix 133.

[0138] 135. The system is a flow cytometer, as described in any one of the appendices 130 to 134. 136. A system as described in any one of the appendices 130 to 135, further comprising a light-gathering system for propagating light to a light-detecting system. 137. The light-gathering component is the system described in Appendix 136, including an optical fiber. 138. The focusing component is an optical fiber relay bundle, as described in Appendix 136. 139. The light-gathering component is a free-space light relay system, as described in Appendix 136.

[0139] 140. Method, This includes detecting light from a flow stream using a light detection system, The light detection system is Three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, One or more photodetector modules that are optically connected to each wavelength separator. Each photodetector module includes, Multiple photodetectors, An optical component configured to transmit light having a predetermined subspectral range to a photodetector, Methods that include... 141. The method according to Appendix 140, further comprising irradiating a sample in a flow stream within an interrogation field with a light source. 142. The flowstream is irradiated with a light source with a wavelength of 200 nm to 800 nm, as described in Appendix 140 or 141. 143. The light source is a laser, as described in Appendix 141 or 142. 144. The method according to any one of the appendices 141 to 143, wherein light from a flowstream is transmitted to a photodetector having a light-collecting component.

[0140] 145. The light-gathering component is the method described in Appendix 144, including an optical fiber. 146. The light-gathering component is the method described in Appendix 145, including an optical fiber repeater bundle. 147. The light-gathering component is the method described in Appendix 144, including a free-space light relay system.

[0141] 148. A method, This includes detecting light from a flow stream using a light detection system, The light detection system is A wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, It is configured to receive light from each of the first, second, and third predetermined spectral ranges. The first, second, and third optical detection modules and The first, second, and third photodetector modules each include, Multiple photodetectors, An optical component configured to transmit light having a predetermined subspectral range to a photodetector, Methods that include... 149. The method according to Appendix 148, further comprising irradiating a sample in a flow stream within an interrogation field with a light source. 150. Flowstream is irradiated with a light source with a wavelength of 200 nm to 800 nm, as described in Appendix 148 or 149. 151. The light source is a laser, as described in Appendix 149 or 150. 152. The method according to any one of the appendices 148 to 151, wherein light from a flowstream is transmitted to a photodetector having a light-collecting component.

[0142] 153. The light-gathering component is the method described in Appendix 152, including an optical fiber. 154. The light-gathering component is the method described in Appendix 153, including an optical fiber repeater bundle. 155. The light-gathering component is the method described in Appendix 152, including a free-space light relay system.

[0143] 156. A method, This includes detecting light from a flow stream using a light detection system, The light detection system is A wavelength separator configured to generate light in first, second, and third predetermined spectral ranges from a light source, First, second, and third photodetector modules configured to receive light from each of the first, second, and third predetermined spectral ranges, respectively. Includes, Each of the first, second, and third photodetector modules is configured to generate light in multiple subspectral ranges. Light in each subspectral range exhibits less than 20% optical loss compared to light from the light source. method. 157. The method described in Appendix 156, wherein the light in each subspectral range exhibits an optical loss of 15% or less compared to the light from the light source. 158. The method according to Appendix 156 or 157, wherein the first, second, and third photodetector modules are configured to generate light in 20 or more subspectral ranges. 159. The method according to Appendix 156 or 157, wherein the first, second, and third photodetector modules each include a plurality of dichroic mirrors and are configured to generate light in 20 or more subspectral ranges with 10 or fewer reflections. 160. The method according to any one of the appendices 156 to 159, further comprising irradiating a sample in a flow stream within an interrogation field with a light source.

[0144] 161. Flowstream is irradiated with a light source with a wavelength of 200 nm to 800 nm, according to any one of the methods described in Appendix 156 to 160. 162. The light source is a laser, as described in Appendix 160 or 161. 163. The method according to any one of the appendices 156 to 162, wherein light from a flowstream is transmitted to a photodetector having a light-collecting component. 164. The light-gathering component is the method described in Appendix 163, including an optical fiber. 165. The optical focusing component is the method described in Appendix 163, including an optical fiber repeater bundle. 166. The light-gathering component is the method described in Appendix 163, including a free-space light relay system.

[0145] 167. It is a kit, Wavelength separator and Multiple photodetectors, Optical components and A kit that includes this. 168. The wavelength separator is configured to generate light of first, second, and third predetermined spectral ranges from a light source, as described in Appendix 167. 169. The wavelength separator is a prism, as described in the kit in Appendix 168. 170. The wavelength separator is a diffraction grating, as described in the kit in Appendix 168. 171. A kit as described in Appendix 167, including three or more wavelength separators.

[0146] 172. The wavelength separator is a dichroic mirror, as described in the kit in Appendix 171. 173. A kit containing multiple photomultiplier tubes, as described in any one of the appendices 167-172. 174. A kit containing multiple avalanche photodiodes, as described in any one of the appendices 167-172. 175. A kit that includes a mount, as described in any one of the appendices 167-174.

[0147] Although the above invention has been described in some detail by examples and illustrations for the sake of clear understanding, it will be readily apparent to those skilled in the art that certain changes and modifications can be made in light of the teachings of the present invention without departing from the spirit or scope of the appended claims.

[0148] Therefore, the above merely illustrates the principles of the present invention. Those skilled in the art will understand that various arrangements embodying the principles of the present invention and falling within its spirit and scope can be devised, although not expressly described or shown herein. Furthermore, all examples and conditional language described herein are intended primarily to help the reader understand the principles of the present invention and the concepts to which the inventors contribute to the advancement of the art, and should be construed as not being limited to such specifically described examples and conditions. Furthermore, all descriptions herein describing the principles, aspects, and embodiments of the present invention, and specific examples thereof, are intended to encompass both their structural and functional equivalents. In addition, such equivalents are intended to include both currently known equivalents and equivalents to be developed in the future, i.e., any elements developed to perform the same function regardless of their structure. Furthermore, nothing disclosed herein, whether or not such disclosure is expressly described in the claims, is intended to be dedicated to the public.

[0149] Accordingly, the scope of the present invention is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the present invention are embodied in the appended claims. In the claims, Section 112(f) or Section 112(6) of the U.S. Patent Act is expressly defined as being invoked for the limitation in the claims only if the exact phrase “means for” or “steps for” is found at the beginning of such limitation in the claims, and if such exact phrase is not used in the limitation in the claims, Section 112(f) or Section 112(6) of the U.S. Patent Act is not invoked.

[0150] Cross-reference of related applications In accordance with Section 119(e) of the United States Patent Act, this application claims priority on the filing date of U.S. Provisional Patent Application No. 62,971,840, filed on 7 February 2020, the disclosure of which is incorporated herein by reference.

Claims

[Claim 1] A light detection system, Three or more wavelength separators, each configured to allow light having a predetermined spectral range to pass through, One or more photodetector modules that are optically connected to each wavelength separator. Each photodetector module includes, Multiple photodetectors, An optical component configured to transmit light having a predetermined subspectral range to the photodetector, A light detection system, including a light detection system.

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