Optical detection system and method for a flow cytometer - Patents.com

Abstract

An optical detection system and optical detection method for a flow cytometer are provided. The optical detection system includes a beam separation device and a plurality of wavelength division multiplexing devices. The beam separation device is configured to separate a beam to be processed by the flow cytometer into a plurality of first beams having individual wavelength ranges that are either non-overlapping or partially overlapping with each other. Each of the plurality of wavelength division multiplexing devices is configured to receive an individual one of the plurality of first beams. The plurality of first beams are parallel to each other when received by the plurality of wavelength division multiplexing devices. Each of the plurality of wavelength division multiplexing devices includes a plurality of optical detection devices configured to detect a portion of the individual first beam.

Description

[Technical field]

[0001] (Field) The present disclosure relates to optical detection systems and methods for flow cytometers. [Background technology]

[0002] (background) Optical detection technology is applied in many fields. For example, optical detection technology is applied to detect the volume, morphology, properties, and the like of an object. For example, optical detection technology can be used in flow cytometry to detect the volume, morphology, properties, and the like of a microparticle. Summary of the Invention [Means for solving the problem]

[0003] (summary) A brief summary of the present disclosure is provided below to provide a basic understanding of certain aspects of the disclosure. It should be understood, however, that the summary is not an exhaustive summary of the disclosure. It is not intended to determine essential or critical portions of the disclosure, nor is it intended to limit the scope of the disclosure. Its purpose is to present some concepts of the disclosure only in a simplified form as a prelude to the more detailed description that is presented later.

[0004] According to an aspect of the present disclosure, an optical detection system for a flow cytometer is provided. The optical detection system includes a beam separation device and a plurality of wavelength division multiplexing devices. The beam separation device is configured to separate a beam to be processed by the flow cytometer into a plurality of first beams having individual wavelength ranges that are either non-overlapping with each other or partially overlapping with each other. Each of the plurality of wavelength division multiplexing devices is configured to receive an individual one of the plurality of first beams. The plurality of first beams are parallel to each other when received by the plurality of wavelength division multiplexing devices. Each of the plurality of wavelength division multiplexing devices includes a plurality of optical detection devices configured to detect a portion of the individual first beam.

[0005] According to another aspect of the present disclosure, an optical detection method for a flow cytometer is provided, the optical detection method including: separating, by a beam separation device, a beam to be processed by the flow cytometer into a plurality of first beams having individual wavelength ranges that are either non-overlapping or partially overlapping with each other; and multiplexing and detecting, by a plurality of wavelength division multiplexing devices, the plurality of first beams, respectively. Each wavelength division multiplexing device includes a plurality of optical detection devices configured to detect individual ones of the plurality of first beams, the plurality of first beams being parallel to each other when received by the wavelength division multiplexing device.

[0006] According to other aspects of the present disclosure, there is further provided computer program code and computer program products for implementing methods according to the present disclosure, and computer readable storage media having recorded thereon computer program code for implementing methods according to the present disclosure.

[0007] Other aspects of the embodiments of the present disclosure are given in the following specification: In order to fully disclose the present disclosure, preferred embodiments are described in detail, without limitation. [Brief description of the drawings]

[0008] The present disclosure may be better understood by reference to the detailed description given below in conjunction with the drawings, in which the same or similar reference numbers are used to represent the same or similar components in the drawings, and the drawings, together with the following detailed description, are incorporated in and form a part of the specification, further illustrating preferred embodiments of the present disclosure and explaining the principles and advantages of the present disclosure.

[0009] [Figure 1] FIG. 1 is a block diagram of an example configuration of an optical detection system for a flow cytometer, according to one embodiment of the present disclosure. [Diagram 2] FIG. 2 is a 3D layout showing an implementation of an optical detection system for a flow cytometer according to one embodiment of the present disclosure. [Diagram 3] FIG. 3 is a top view showing an implementation of an optical detection system for a flow cytometer according to an embodiment of the present disclosure. [Figure 4] FIG. 4 is a side view showing an implementation of an optical detection system for a flow cytometer according to an embodiment of the present disclosure. [Diagram 5] 5A and 5B are diagrams illustrating an implementation of a beam separation device according to an embodiment of the present disclosure. [Figure 6] 6A, 6B, and 6C are diagrams illustrating another implementation of a beam separating device according to an embodiment of the present disclosure. [Figure 7] FIG. 7 is a top view showing another implementation of an optical detection system for a flow cytometer according to an embodiment of the present disclosure. [Figure 8] FIG. 8 is a schematic diagram of an implementation of a wavelength division multiplexing unit according to an embodiment of the present disclosure. [Figure 9] 9A and 9B are schematic diagrams illustrating an example of a support base included in a wavelength division multiplexing device according to an embodiment of the present disclosure, and an arrangement of wavelength division multiplexing units on a support base according to an embodiment of the present disclosure. [Figure 10A]FIG. 10A illustrates another example of a support base included in a wavelength division multiplexing device according to an embodiment of the present disclosure. [Figure 10B] 10B-10D are schematic diagrams illustrating an arrangement of wavelength division multiplexing units on a supporting base according to one embodiment of the present disclosure. [Figure 10C] 10B-10D are schematic diagrams illustrating an arrangement of wavelength division multiplexing units on a supporting base according to one embodiment of the present disclosure. [Figure 10D] 10B-10D are schematic diagrams illustrating an arrangement of wavelength division multiplexing units on a supporting base according to one embodiment of the present disclosure. [Figure 11] FIG. 11 is a schematic diagram of another implementation of a wavelength division multiplexing unit in accordance with an embodiment of the present disclosure. [Figure 12] FIG. 12 is a schematic diagram of yet another implementation of a wavelength division multiplexing unit in accordance with an embodiment of the present disclosure. [Figure 13] FIG. 13 is a flow chart showing an exemplary flow of an optical detection method for flow cytometry according to an embodiment of the present disclosure. [Figure 14] FIG. 14 is a block diagram of an exemplary structure of a personal computer applicable to embodiments of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Detailed Description of the Embodiments Exemplary embodiments of the present disclosure are described below in conjunction with the drawings. For the sake of brevity and clarity, not all features of an actual embodiment are described herein. However, it should be understood that numerous embodiment-specific decisions, for example according to system and business-related constraints, should be made when developing any such actual embodiment to achieve the developer's specific targets. These constraints may vary with different embodiments. Moreover, it should be understood that while the development efforts may be complex and time-consuming, such development efforts would be no more than a routine task for those skilled in the art having the benefit of the present disclosure.

[0011] The terms "first", "second", and the like may be used to describe various elements herein, but it should be understood that these elements should not be limited by these terms. These terms are used only to distinguish one element from another. For example, a first element may be referred to as a second element, and similarly, a second element may be referred to as a first element, without departing from the scope of the present disclosure.

[0012] In addition, in this specification and drawings, there are also cases where multiple components with basically the same functional configuration are distinguished by adding different letters after the same reference number. For example, the wavelength division multiplexing unit 1046 may be referred to as a wavelength division multiplexing unit 1046a, a wavelength division multiplexing unit 1046b, and a wavelength division multiplexing unit 1046c, as necessary. However, when it is unnecessary to particularly distinguish multiple components with basically the same functional configuration from each other, only the same reference number is required to present the components. For example, when it is unnecessary to distinguish the wavelength division multiplexing unit 1046a, the wavelength division multiplexing unit 1046b, and the wavelength division multiplexing unit 1046c from each other, the wavelength division multiplexing unit 1046a, the wavelength division multiplexing unit 1046b, and the wavelength division multiplexing unit 1046c are simply referred to as the wavelength division multiplexing unit 1046.

[0013] It should also be noted here that in order to avoid obscuring the present disclosure due to unnecessary details, only device structures and / or processing steps that are closely related to the solutions according to the present disclosure are shown in the drawings, and other details that are not closely related to the present disclosure are omitted.

[0014] Embodiments according to the present disclosure are described in detail below in conjunction with the drawings.

[0015] First, an example implementation of an optical detection system for a flow cytometer according to an embodiment of the present disclosure will be described with reference to FIGS. 1-7. FIG. 1 is a block diagram of an example configuration of an optical detection system 100 for a flow cytometer according to an embodiment of the present disclosure. FIGS. 2, 3, and 4 are 3D layout, top view, and side view, respectively, illustrating an implementation of an optical detection system 100 for a flow cytometer according to an embodiment of the present disclosure. FIGS. 5A and 5B are diagrams illustrating an implementation of a beam separation device according to an embodiment of the present disclosure. FIGS. 6A, 6B, and 6C are diagrams illustrating another implementation of a beam separation device according to an embodiment of the present disclosure. FIG. 7 is a top view illustrating another implementation of an optical detection system for a flow cytometer according to an embodiment of the present disclosure.

[0016] As shown in FIGS. 1-4, an optical detection system 100 for a flow cytometer according to an embodiment of the present disclosure includes a beam separation device 102 and a plurality of wavelength division multiplexing devices 104.

[0017] The beam separation device (also referred to as "spectral band separation device") 102 is configured to separate a beam to be processed by a flow cytometer (hereinafter also referred to as "beam to be processed") into a plurality of first beams having distinct wavelength ranges that either do not overlap with each other or partially overlap with each other. For example, the beam to be processed may be transmitted to the beam separation device 102 through an optical fiber. In addition, for example, the beam to be processed may be transmitted to the beam separation device 102 through free space using a coupled lens system.

[0018] As an example, the beam splitting device 102 includes a long-pass filter or a short-pass filter. For example, as shown in Fig. 2 and Fig. 4, the beam splitting device 102 may be arranged at 45 degrees with respect to the optical axis of the beam to be processed and split the beam to be processed into two first beams propagating along the x-axis direction and the z-axis direction, respectively. For example, in the case where the beam to be processed has a wavelength range of 350 nm to 950 nm, the beam splitting device 102 splits the beam to be processed into two first beams having a wavelength range of 350 nm to A and a wavelength range of A to 950 nm, respectively. For example, A is greater than or equal to 400 nm and less than or equal to 900 nm. It should be noted that in Fig. 4, the elements surrounded by dotted frames correspond to the two first beams, respectively.

[0019] As another example, the beam splitting device 102 may include a dichroic mirror. For example, as shown in Figures 5A and 5B, the dichroic mirror can reflect visible light (Vis) and infrared light (IR) in one band and transmit ultraviolet light (UV) and visible light in another band to split the beam to be processed into two first beams. In addition, by arranging two or more dichroic mirrors with different optical properties, the beam to be processed is split into three or more first beams. For example, as shown in Figure 6A, by arranging two dichroic mirrors with different optical properties as shown in Figures 6B and 6C, the beam to be processed is split into three first beams.

[0020] In Fig. 2, the grey lines represent the beams. Additionally, in Fig. 5A and Fig. 6A, the arrows represent the beams. As can be understood by those skilled in the art, the actual shape of the beams may differ from that shown in the drawings. Additionally, the relative positions between the first beams are not limited by the drawings.

[0021] Each of the wavelength division multiplexing devices 104 is configured to receive a respective one of the multiple first beams. For example, the wavelength division multiplexing devices 104 may correspond to the first beams in a one-to-one relationship.

[0022] Each one of the wavelength division multiplexing devices 104 includes a plurality of photodetection devices 1042 (not shown in FIG. 2 ). The photodetection devices 1042 are configured to detect (e.g., detect the optical intensity of) a portion of the first beam received by the wavelength division multiplexing device 104. For example, for each wavelength division multiplexing device 104, the wavelength division multiplexing device 104 may each perform wavelength division multiplexing on the first beam received by the wavelength division multiplexing device 104 to obtain a plurality of second beams and transmit the plurality of second beams to the plurality of photodetection devices 1042. In Figures 2-4, the wavelength division multiplexing device 104 is shown to include a wavelength division multiplexing unit 1046b, shown in Figure 11, and to perform wavelength division multiplexing on the first beam received by the wavelength division multiplexing device, but it should be noted that those skilled in the art may adopt another wavelength division multiplexing unit according to actual needs, such as the wavelength division multiplexing unit 1046a, shown in Figure 8, the wavelength division multiplexing unit 1046c, shown in Figure 12, and equivalents.

[0023] As an example, the multiple second beams may be focused onto multiple photodetector devices 1042 through multiple second aspheric lenses 122 as shown in FIGS.

[0024] The light detection device 1042 may be, for example, an avalanche photodiode (APD).

[0025] The multiple first beams may be parallel to each other when received by the multiple wavelength division multiplexing devices 104. In this case, for example, the photodetector devices 1042 included in the multiple wavelength division multiplexing devices 104 may be arranged coplanarly to facilitate, for example, temperature control of the photodetector devices 1042 and layout of signal processing circuits and wires.

[0026] 4, the transmission path of at least one of the plurality of first beams may be changed by the second mirror 118 such that the plurality of first beams are transmitted parallel to the plurality of wavelength division multiplexing devices 104. For example, the second mirror 118 may change the transmission path of the first beam by totally reflecting the first beam. The second mirror 118 may be a plane mirror.

[0027] As another example, multiple first beams may be transmitted to multiple wavelength division multiplexing devices 104 while not being parallel to each other. For example, in the optical detection system 100b shown in FIG. 7, two beams may be transmitted to separate wavelength division multiplexing devices 104 with a 90 degree angle between the two beams. In addition, in practical applications, the angle between the beams may be changed by configuring the beam splitting device 102 as needed. For example, the angle between the two beams may range from 15 degrees to 165 degrees.

[0028] At present, flow cytometry is evolving from traditional multi-color fluorescence channels to high-channel fluorescence full spectrum. With fluorescence full spectrum, more fluorescence information is obtained, and more (up to 40 or more or more channels) fluorescein and other information are also analyzed simultaneously, so as to obtain more sample expression information and thereby achieve sensitive and accurate test results. To achieve this goal, high-density channel acquisition of fluorescence spectrum is required to reflect the spectral characteristics of fluorescence.

[0029] As described above, in the optical detection system 100 according to the embodiment of the present disclosure, the beam to be processed (e.g., including the fluorescent signal) is separated into a plurality of first beams by the beam separation device 102, and then the first beams are multiplexed and detected by the wavelength division multiplexing device 104, so that the number of optical channels can be easily increased (e.g., increased to 20 or more) and good optical properties are maintained. In addition, compared with the case where the beam to be processed is directly multiplexed and detected by the wavelength division multiplexing device, by using the optical detection system 100, the optical path length of at least one or more channels corresponding to the first beam and the number of optical elements through which the first beam passes are reduced, thereby improving the optical transmission efficiency of at least one or more channels and reducing the size of the wavelength division multiplexing device 104.

[0030] In addition, the optical detection system 100 may be compatible with a dual-port wavelength division multiplexing structure. Two excitation sources (examples of objects that generate beams to be processed) with a small number of channels (e.g., less than or equal to 10) may share one dual-port wavelength division multiplexing structure, which saves costs.

[0031] As an example, the multiple first beams may be transmitted in a layered layout in a vertical direction (e.g., z-axis direction, as shown in FIG. 2 ), such that the size of the wavelength division multiplexing device 104 may be further reduced. According to an embodiment of the present disclosure, the beam to be processed by the flow cytometer may include a side scattered light signal and a fluorescent signal. In this case, as shown in FIGS. 3 and 4 , the light detection system may further include a multiple side scattered light signal separation device 106. Each of the multiple side scattered light signal separation devices 106 is configured to separate the side scattered light signal from a respective one of the multiple first beams (i.e., the first beam transmitted to the side scattered light signal separation device 106) and transmit the separated respective first beam to a respective one of the multiple wavelength division multiplexing devices (e.g., a wavelength division multiplexing device configured to multiplex and detect the separated respective first beams), such that the influence of the side scattered light signal on the fluorescent signal is reduced, thereby improving the signal-to-noise ratio of the fluorescent signal. For example, a plurality of side-scatter light signal separation devices 106 may correspond in a one-to-one relationship to a plurality of first beams.

[0032] As an example, the beam to be processed may comprise a beam collected from a particle, and the fluorescence signal may comprise a fluorescence signal emitted from the particle.

[0033] According to certain embodiments of the present disclosure, each side scattered light signal separation device 106 may include a first side scattered light signal separation unit 1062 and a second side scattered light signal separation unit 1064 (not shown in FIG. 2 ). The first side scattered light signal separation unit 1062 may be configured to separate the side scattered light signal from the individual first beams, and the second side scattered light signal separation unit 1064 may be configured to further separate the side scattered light signal from the individual first beams separated by the first side scattered light signal separation unit 1062 and transmit the separated individual first beams to individual ones of the multiple wavelength division multiplexing devices 104, such that, for example, the signal-to-noise ratio of the fluorescence signal is further improved.

[0034] For example, the first side-scattered light signal separation unit 1062 may include a long band-pass filter, for example, the long band-pass filter may be positioned at 20 degrees to 50 degrees with respect to the optical axis of the corresponding first beam, and may reflect the side-scattered light signal and transmit the fluorescent light signal.

[0035] As an example, the second side-scattered light signal separation unit 1064 may include a long band-pass filter, for example, the long band-pass filter may be positioned at -5 degrees or 5 degrees relative to the optical axis of the corresponding first beam, to deeply isolate the residual side-scattered light signals in the multiple first beams from the first side-scattered light signal separation unit 1062.

[0036] For example, as shown in FIG. 3, FIG. 4, and FIG. 7, the light detection system 100 may further include a plurality of attenuation devices 108, a plurality of first bandpass filters 110, and a plurality of first light detection units 112. Each of the plurality of attenuation devices 108 may be configured to attenuate a side scattered light signal separated by a first side scattered light signal separation unit 1062 corresponding to the attenuation device 108, and prevent the side scattered light signal from saturating the first light detection unit 112. Each of the plurality of first bandpass filters 110 may be configured to perform bandpass filtering on the side scattered light signal attenuated by the attenuation device 108 corresponding to the first bandpass filter 110. Each of the plurality of first light detection units 112 may be configured to detect (e.g., detect the intensity of) a side scattered light signal filtered by the first bandpass filter 110 corresponding to the first light detection unit 112, for example, so that the size of a microparticle to be screened by a flow cytometry method is detected.

[0037] For example, the multiple attenuation devices 108 may have the same attenuation coefficient, or at least one of the multiple attenuation devices 108 has an attenuation coefficient that is different from that of the other attenuation devices 108. Similarly, the multiple first bandpass filters 110 may have the same optical properties, or at least one of the multiple first bandpass filters 110 has optical properties that are different from that of the other first bandpass filters 110.

[0038] As an example, when the beam to be processed is split into two first beams by the beam splitting device 102, the intensity of the side-scattered light signal contained in one of the two first beams (hereinafter referred to as "SSC1") and the intensity of the side-scattered light signal contained in the other of the two first beams (hereinafter referred to as "SSC2") are b% and 1-b%, respectively, of the intensity of the side-scattered light signal contained in the beam to be processed (hereinafter referred to as "SSC"), where 1≦b≦2. On the one hand, the side-scattered light signal generated by small particles (such as particles with nanometer size) is so weak that it is desired to improve the efficiency of collecting side-scattered light signals from small particles as much as possible. On the other hand, the side-scattered light signal generated by large particles (such as cells with micron size) is so strong that it is desired to reduce the efficiency of collecting side-scattered light signals from large particles, which leads to the light detection device being easily saturated. In the case where SSC1 and SSC2 are configured as described above, for example, SSC1 may be used to detect cells with sizes ranging from 1 μm to 30 μm, and SSC2 may be used to detect small particles with sizes ranging from 80 nm to 1 μm, so that the accuracy of detecting particles is further improved. For example, in the optical detection system 100a shown in FIG. 4, the elements surrounded by the upper dotted frame and the elements surrounded by the lower dotted frame may correspond to the first beam including SSC1 and the first beam including SSC2, respectively. In addition, for example, in the optical detection system 100b shown in FIG. 7, the elements surrounded by the dotted frame indicated by the reference number SSC1 may be used for the transmission and detection of SSC1, and the elements surrounded by the dotted frame indicated by the reference number SSC2 may be used for the transmission and detection of SSC2.

[0039] 2-4 and 7, the light detection system 100 may further include a plurality of first mirrors 114. Each of the plurality of first mirrors 114 is configured to change a transmission path of a side-scattered light signal separated by a first side-scattered light signal separation unit 1062 corresponding to the first mirror 114, and transmit the side-scattered light signal to a respective one of the plurality of attenuation devices 108.

[0040] For example, as shown in FIG. 3, the first mirror 114 may be arranged at 20 degrees to 50 degrees with respect to the optical axis of the side scattered light signal from the first side scattered light signal separation unit 1062 corresponding to the first mirror 114, so as to change the transmission path of the side scattered light signal so that the side scattered light signal and the first beam separated by the first side scattered light signal separation unit 1062 are transmitted in parallel, and may reflect the side scattered light signal. In this case, for example, the multiple first light detection units 112 and the multiple light detection devices 1042 may be arranged on the same plane, for example, to facilitate temperature control for the multiple first light detection units 112 and the multiple light detection devices 1042 and to facilitate the layout of the signal processing circuitry and wires. For example, the multiple first light detection units 112 and the multiple light detection devices 1042 may be arranged with reference to a virtual reference plane represented by a dotted line in FIG. 3 so that the multiple first light detection units 112 and the multiple light detection devices 1042 are accurately arranged on the same plane.

[0041] 3, 4, and 7, the light detection system 100 may further include a plurality of first aspheric lenses 120 disposed between the plurality of first bandpass filters 110 and the plurality of first light detection units 112. Each of the plurality of first aspheric lenses 120 is configured to focus a side-scattered light signal, which is filtered by the first bandpass filter 110 corresponding to the first aspheric lens 120, onto a respective one of the plurality of first light detection units 112.

[0042] For example, as shown in Figures 3, 4, and 7, the optical detection system 100 may further include an achromatic lens 116 configured to collimate the beam to be processed and transmit the collimated beam to the beam separation device 102. For example, the beam to be processed is collected through optical fiber coupling. In addition, for example, the beam to be processed may be transmitted to the achromatic lens 116 through free space using a coupled lens system.

[0043] A wavelength division multiplexing unit according to an embodiment of the present disclosure is described in detail below with reference to FIGS.

[0044] FIG. 8 is a schematic diagram of an implementation of a wavelength division multiplexing unit according to an embodiment of the present disclosure. As shown in FIG. 8, the wavelength division multiplexing unit 1046a may include a plurality of mirrors 1046-M and a plurality of second bandpass filters 1046-L disposed opposite the plurality of mirrors 1046-M. Each mirror 1046-M may be configured to reflect a beam transmitted from a respective one of the plurality of second bandpass filters 1046-L to the mirror 1046-M. For example, each mirror 1046-M may include a concave portion (e.g., a fovea portion) C disposed between a pair of planar portions P. Each concave portion C is configured to reflect a beam transmitted to the concave portion C from a corresponding one of the plurality of second bandpass filters 1046-L to collimate the beam. In addition, each second bandpass filter 1046-L may be configured to transmit light having a wavelength within a wavelength range corresponding to the second bandpass filter 1046-L and reflect other light. Thus, the beam propagates in a zigzag manner between the plurality of mirrors 1046-M and the plurality of second bandpass filters 1046-L to separate a corresponding light beam (e.g., a first beam) into a plurality of second beams, e.g., ten second beams corresponding to channel #1-channel #10 as shown in FIG. 8. In addition, by disposing the concave portion C between the flat portion P, the divergent light transmitted over a distance is collimated to achieve long-distance transmission, which leads to detecting more channels. Note that although FIG. 8 shows that adjacent mirrors 1046-M are spaced apart, the plurality of mirrors 1046-M may be disposed without a space between them, if desired. Additionally, for example, multiple mirrors 1046-M may be arranged as a single mirror 1046-M with one or more planar portions P and concave portions C that extend the length of wavelength division multiplexing unit 1046a.

[0045] In some embodiments, the directivity of the beam may be adjusted by adjusting the relative position between the second bandpass filter 1046-L and the corresponding mirror 1046-M, e.g., the distance between the center of the second bandpass filter 1046-L and the center of the corresponding mirror 1046-M, in the direction along which the second bandpass filter 1046-L is arranged to further ensure that the beam propagates in a zigzag manner.

[0046] As an example, the wavelength division multiplexing device 104 may further include a support base 1048a, as shown in Fig. 9A. For example, as shown in Fig. 9B, a plurality of mirrors (also referred to as a "mirror array") may be disposed on a first surface SA1 of the support base 1048a, and a plurality of second bandpass filters (also referred to as a "second bandpass filter array") may be disposed on a second surface SA2 of the support base 1048a, opposite and parallel to the first surface SA1, to ensure parallelism between the mirror array and the second bandpass filter array and improve light transmission efficiency.

[0047] For example, as shown in Fig. 9B, a plurality of mirrors 1046-M and a plurality of second bandpass filters 1046-L may be disposed over the first surface SA1 and the second surface SA2, respectively, so that the beam is transmitted, rather than passing, through the interior of the support base 1048a, to avoid the interface light loss (Ferrer reflection loss) caused by the light transmission through the interior of the support base 1048a, thereby further improving the light transmission efficiency. In addition, the light scattering caused by crystallization or bubbles in the material of the support base 1048a is avoided, so that the optical signal crosstalk between different channels is reduced.

[0048] As another example, the wavelength division multiplexing device 104 may further include a support base 1048b with a hollow interior, as shown in FIG. 10A. For example, as shown in FIG. 10B-FIG. 10D, the mirrors 1046-M may be disposed on a first surface SB1 of the support base 1048b, and the second bandpass filters 1046-L may be disposed on a second surface SB2 of the support base 1048b, which is opposite and parallel to the first surface SB1, to ensure parallelism between the mirrors 1046-M and the second bandpass filters 1046-L and improve light transmission efficiency. In addition, in this case, the beam is transmitted through the hollow interior, which avoids interface light loss, thereby further improving light transmission efficiency. Furthermore, light scattering caused by crystallization or bubbles inside the material of the support base 1048b is avoided, so that optical signal crosstalk between different channels is reduced. Additionally, the hollow interior significantly reduces the bulk of the support base 1048b while ensuring parallelism and structural strength.

[0049] For example, the same material as the support base 1048b or any other material with a refractive index greater than 1, such as glass, polymers, and fluids, may be filled within the hollow interior of the support base 1048b.

[0050] For example, the two wavelength division multiplexing devices 104 may share one support base 1048. For example, in the embodiment shown in Figures 9B and 10B, the upper mirror array and the second bandpass filter array correspond to one of the two wavelength division multiplexing devices 104, and the lower mirror array and the second bandpass filter array correspond to the other of the two wavelength division multiplexing devices 104, such that, for example, the size and weight of the wavelength division multiplexing devices 104 may be reduced.

[0051] For example, wavelength division multiplexing units 1046 included in multiple wavelength division multiplexing devices may be arranged as one body.

[0052] The support base 1068 may be made from a transparent or non-transparent material. For example, the support base 1068 may be made from metal, glass, and / or polymer.

[0053] 11 is a schematic diagram of another implementation of a wavelength division multiplexing unit according to an embodiment of the present disclosure. The wavelength division multiplexing unit 1046b shown in FIG. 11 differs from the wavelength division multiplexing unit 1046a shown in FIG. 8 in that the mirror 1046-M is replaced with a plurality of plane mirrors 1046-P and a plurality of concave mirrors 1046-C arranged alternately. Similar to the planar portion P of the mirror 1046-M, each plane mirror 1046-P may be configured to reflect a beam transmitted through the plane mirror 1046-P from a corresponding second bandpass filter 1046-L of the plurality of second bandpass filters 1046-L. In addition, similar to the concave portion C of the mirror 1046-M, each concave mirror 1046-C may be configured to reflect the beam transmitted to the concave mirror from the second bandpass filter 1046-L corresponding to the concave mirror of the plurality of second bandpass filters 1046-L to collimate the beam. Although FIG. 11 shows one flat mirror 1046-P being disposed between two concave mirrors 1046-C, two or more flat mirrors 1046-P may be disposed between two concave mirrors 1046-C as required. For example, as shown in FIG. 3 and FIG. 7, two flat mirrors 1046-P are disposed between two concave mirrors 1046-C.

[0054] 12 is a schematic diagram of yet another implementation of a wavelength division multiplexing unit according to an embodiment of the present disclosure. The wavelength division multiplexing unit 1046c may include a plurality of third bandpass filters 1046-J and a plurality of fourth bandpass filters 1046-K disposed opposite to the plurality of third bandpass filters 1046-J. Each of the third bandpass filters 1046-J and the fourth bandpass filters 1046-K transmits light having a wavelength within a wavelength range corresponding to the bandpass filter and reflects other light. For example, as shown in FIG. 12, a concave mirror may be disposed between some of the plurality of third bandpass filters 1046-J to collimate the beam transmitted to the concave mirror from the corresponding fourth bandpass filter 1046-K of the plurality of fourth bandpass filters 1046-K. Compared with the wavelength division multiplexing units 1046a and 1046b, the wavelength division multiplexing unit 1046c further increases the number of fluorescence channels. It should be noted that although only one concave mirror is arranged as shown in FIG. 12, two or more concave mirrors may be arranged according to actual needs. For example, the third band pass filter 1046-J and the concave mirror may be arranged alternately in the same manner as that of the plane mirror 1046-P and the concave mirror 1046-C as described above with reference to FIG. 11. In addition, the position of the concave mirror is not limited by the drawing.

[0055] For example, wavelength division multiplexing units 1046b and 1046c may be arranged in a manner similar to the arrangement of wavelength division multiplexing unit 1046a, described with reference to Figures 9A-10D, which are not repeated here.

[0056] In the wavelength division multiplexing unit 1046 described above with reference to Figures 8-12, the number of fluorescence channels can be easily increased by increasing the number of corresponding elements (e.g., mirror 1046-M and second bandpass filter 1046-L shown in Figure 8).

[0057] The above mainly describes an optical detection system for flow cytometry, however the optical detection system may also be applied to other devices such as a sorting device.

[0058] According to an embodiment of the present disclosure, an optical detection method for flow cytometry is provided. Figure 13 is a flow chart of an optical detection method 1000 for flow cytometry according to an embodiment of the present disclosure.

[0059] As shown in FIG. 13, the optical detection method 1000 includes a beam separation step S1020 and a beam detection step S1040.

[0060] In a beam separation step S1020, a beam to be processed by flow cytometry (hereinafter also referred to as a "beam to be processed") is separated by a beam separation device into a plurality of first beams having individual wavelength ranges that are either non-overlapping or partially overlapping with each other. For example, the beam to be processed may be separated into a plurality of first beams by the beam separation device 102 described above.

[0061] In a beam detection step S1040, the first beams are detected by a plurality of optical detection devices included within a plurality of wavelength division multiplexing devices, such as wavelength division multiplexing device 104. For example, the first beams are parallel to one another when received by the wavelength division multiplexing device.

[0062] Using the optical detection method 1000 according to an embodiment of the present disclosure, a beam to be processed is separated into multiple first beams by a beam separating device, and then the multiple first beams are multiplexed and detected by a wavelength division multiplexing device so that the number of optical channels can be easily increased (e.g., increased to 20 or more) and good optical performance is maintained.

[0063] As an example, the beam to be processed may include a side-scattered light signal and a fluorescent light signal. In this case, the light detection method 1000 may further include separating the side-scattered light signal from the plurality of first beams by a side-scattered light signal separation device and transmitting the separated plurality of first beams to a wavelength division multiplexing device. For example, the side-scattered light signal may be separated from the plurality of first beams by the side-scattered light signal separation device 106, described above.

[0064] For example, the beam to be processed may include a beam collected from a particle, and the fluorescent signal may include a fluorescent signal emitted from the particle.

[0065] For example, the light detection method 1000 may further include attenuating the side scattered light signal obtained through separation by the side scattered light signal separation device by an attenuation device (e.g., attenuation device 108 as shown in Figures 2-4), performing bandpass filtering on the side scattered light signal attenuated by the attenuation device by a first bandpass filter (e.g., first bandpass filter 110 as shown in Figures 2-4), and detecting the side scattered light signal filtered by the first bandpass filter by a first light detection unit (e.g., first light detection unit 112 as shown in Figures 3 and 4), for example to detect the size of an object that is the source of the side scattered light signal.

[0066] As an example, the optical detection method 1000 may further include changing a transmission path of at least one of the multiple first beams by a second mirror such that the multiple first beams are transmitted in parallel.

[0067] Although the functional configuration of the light detection system and the light detection method according to the embodiments of the present disclosure have been described above, it should be noted that the above description is merely illustrative and not restrictive. Those skilled in the art may modify the above embodiments based on the principles of the present disclosure. For example, those skilled in the art may add, delete, or combine functional modules and operations in the above embodiments. Such modifications fall within the scope of the present disclosure.

[0068] Furthermore, it should be noted that details described in the system embodiments may also be applied to the method embodiments, and similarly details described in the method embodiments may also be applied to the system embodiments.

[0069] In addition, a storage medium and a program product are also provided according to the present disclosure. It should be understood that the machine executable instructions in the storage medium and the program product according to the embodiments of the present disclosure may be further configured to implement the above-mentioned light detection method. Therefore, details not described herein may refer to the corresponding parts in the above and will not be repeated here.

[0070] Thus, a storage medium for carrying a program product containing machine-executable instructions is also included within the present disclosure, including, but not limited to, a floppy disk, an optical disk, a magneto-optical disk, a memory card, a memory stick, and the like.

[0071] In addition, it should be noted that the above-mentioned series of processes and devices may also be implemented by software and / or firmware. In the case where the above-mentioned series of processes and devices are implemented by software and / or firmware, the program constituting the software is installed from a storage medium or a network into a computer with a dedicated hardware structure, for example, a general-purpose personal computer 1700 shown in FIG. 14. When various programs are installed in the computer, the computer can perform various functions.

[0072] 14, a central processing unit (CPU) 1701 executes various processes according to a program stored in a read-only memory (ROM) 1702 or a program loaded from a storage portion 1708 into a random access memory (RAM) 1703. Data required when the CPU 1701 executes various processes is also stored in the RAM 1703 as necessary.

[0073] The CPU 1701, the ROM 1702, and the RAM 1703 are connected to one another via a bus 1704. An input / output interface 1705 is also connected to the bus 1704.

[0074] The following parts are connected to the input / output interface 1705: an input part 1706 including a keyboard, a mouse, and the like, an output part 1707 including a display such as a cathode ray tube (CRT) and a liquid crystal display (LCD), a loudspeaker, and the like, a storage part 1708 including a hard disk and the like, and a communication part 1709 including a network interface card such as a local area network (LAN) card, a modem, and the like. The communication part 1709 performs communication operations over a network such as the Internet.

[0075] A driver 1710 may also be connected to the input / output interface 1705, if necessary. A removable medium 1711, such as a magnetic disk, an optical disk, a magneto-optical disk, and a semiconductor memory, is loaded onto the driver 1710, if necessary, so that a computer program that is read from the removable medium 1711 is installed in the storage portion 1708, if necessary.

[0076] When the above series of processes is implemented by software, the programs constituting the software are installed from a network such as the Internet or a storage medium such as the removable medium 1711 .

[0077] Those skilled in the art should understand that the storage medium is not limited to the removable medium 1711 shown in FIG. 14 having the program stored therein, and is distributed separately from the device to provide the program to the user. Examples of the removable medium 1711 include magnetic disks (including floppy disks), optical disks (including compact disk read only memory (CD-ROM) and digital versatile disk (DVD)), magneto-optical disks (including mini disks (MD)), and semiconductor memories. Alternatively, the storage medium may be the ROM 1702, a hard disk included in the storage portion 1708, or the like. The storage medium is distributed to the user together with the device having the program stored therein and in which the storage medium is included.

[0078] Preferred embodiments of the present disclosure are described above with reference to the drawings. However, the present disclosure is not limited to the above embodiments. Those skilled in the art may obtain various modifications and changes within the scope of the appended claims. It should be understood that these modifications and changes necessarily fall within the technical scope of the present disclosure.

[0079] For example, in the above embodiment, multiple functions implemented by one unit may be implemented by separate devices. Alternatively, multiple functions implemented by multiple units in the above embodiment may each be implemented by a separate device. In addition, one of the above functions may be implemented by multiple units. Of course, such a configuration is included in the technical scope of the present disclosure.

[0080] In this specification, the steps described in the flowcharts include processes that are performed not only in chronological order in the order described, but also in parallel or separately, not necessarily in chronological order. Furthermore, steps performed in chronological order may be performed in another order, if appropriate.

Claims

1. 1. An optical detection system for a flow cytometer, comprising: a beam separating device configured to separate the beam to be processed by the flow cytometer into a plurality of first beams having distinct wavelength ranges that either do not overlap with one another or partially overlap with one another; a plurality of wavelength division multiplexing devices, each of the plurality of wavelength division multiplexing devices configured to receive a respective one of the plurality of first beams, the plurality of first beams being parallel to one another when received by the plurality of wavelength division multiplexing devices; Equipped with an optical detection system, wherein each of the plurality of wavelength division multiplexing devices includes a plurality of optical detection devices configured to detect a portion of the respective first beam;

2. the beam to be processed by the flow cytometer includes a side scatter signal and a fluorescence signal; 2. The optical detection system of claim 1, further comprising a plurality of side-scattered optical signal separation devices, each configured to separate a side-scattered optical signal from a respective one of the plurality of first beams and transmit the separated respective first beam through a respective one of the plurality of wavelength division multiplexing devices.

3. Each side scatter light signal separation device is a first side-scattered light signal separation unit configured to separate the side-scattered light signal from the individual first beam; a second side-scattered light signal separation unit configured to further separate the side-scattered light signals from the separated individual first beams and transmit the separated individual first beams through respective ones of the plurality of wavelength division multiplexing devices; The optical detection system of claim 2 , comprising:

4. a plurality of attenuation devices, each configured to attenuate the side-scattered light signal separated by the first side-scattered light signal separation unit included in a respective one of the plurality of side-scattered light signal separation devices; a plurality of first bandpass filters, each configured to perform bandpass filtering on the side-scattered light signal attenuated by a respective one of the plurality of attenuation devices; a plurality of first light detection units, each configured to detect the side-scattered light signal filtered by a respective one of the plurality of first band-pass filters; The optical detection system of claim 3 further comprising:

5. 5. The optical detection system of claim 4, further comprising a plurality of first mirrors, each configured to change the transmission path of the side-scattered light signal separated by the first side-scattered light signal separation unit included in a respective one of the plurality of side-scattered light signal separation devices, and to transmit the side-scattered light signal to a respective one of the plurality of attenuation devices.

6. splitting the beam to be processed by the flow cytometer into a plurality of first beams includes splitting the beam to be processed by the flow cytometer into two first beams; 6. The optical detection system of claim 5, wherein the intensity of the side scattered light signal contained in one of the two first beams and the intensity of the side scattered light signal contained in the other of the two first beams are b% and 1-b%, respectively, of the intensity of the side scattered light signal contained in the beam to be processed by the flow cytometer, where 1≦b≦2.

7. 5. The optical detection system of claim 4, further comprising a plurality of first aspherical lenses disposed between the plurality of first bandpass filters and the plurality of first optical detection units, each configured to focus the side-scattered light signal filtered by a respective one of the plurality of first bandpass filters onto a respective one of the plurality of first optical detection units.

8. (i) a chromatic aberration correction lens configured to collimate the beam to be processed by the flow cytometer and transmit the collimated beam to the beam separation device; (ii) a second mirror configured to change the transmission path of at least one of the plurality of first beams such that the plurality of first beams is transmitted parallel to the plurality of wavelength division multiplexing devices; or (iii) a plurality of second aspheric lenses configured to focus a plurality of second beams obtained by separating the respective first beams by respective ones of the plurality of wavelength division multiplexing devices onto the plurality of photodetector devices within the respective wavelength division multiplexing devices. The optical detection system of claim 1 further comprising:

9. The optical detection system of claim 1 , wherein the beam separation device comprises a long-pass filter or a short-pass filter.

10. Each wavelength division multiplexing device comprises a wavelength division multiplexing unit including a plurality of mirrors and a plurality of second bandpass filters disposed opposite the plurality of mirrors; The optical detection system of claim 1 , wherein each mirror is configured to reflect a beam transmitted therethrough from a respective one of the plurality of second bandpass filters.

11. (i) each mirror comprises a concave portion disposed between a pair of planar portions, each concave portion configured to reflect a beam transmitted through said concave portion from a corresponding one of said plurality of second bandpass filters so as to collimate said beam, and said plurality of mirrors are arranged as a single body; or (ii) each wavelength division multiplexing device further comprises a support base, the plurality of mirrors being disposed on a first surface of the support base, the plurality of second bandpass filters being disposed on a second surface of the support base that is opposite and parallel to the first surface, and the support base having a hollow interior.

12. Each wavelength division multiplexing device comprises a wavelength division multiplexing unit including a plurality of alternately arranged flat mirrors and a plurality of concave mirrors, and a plurality of second band pass filters arranged opposite the plurality of flat mirrors and the plurality of concave mirrors; each plane mirror configured to reflect a beam transmitted through the plane mirror from a corresponding one of the plurality of second band-pass filters; 2. The optical detection system of claim 1, wherein each concave mirror is configured to reflect a beam transmitted therethrough from a corresponding one of the plurality of second bandpass filters so as to collimate the beam.

13. The optical detection system of claim 1 , wherein the plurality of optical detection devices are arranged on the same plane.

14. each wavelength division multiplexing device comprises a wavelength division multiplexing unit including a plurality of third band pass filters and a plurality of fourth band pass filters disposed opposite to the plurality of third band pass filters; the wavelength division multiplexing device further comprising a support base with a hollow interior; 2. The optical detection system of claim 1, wherein the plurality of third bandpass filters are disposed on a first surface of the support base, and the plurality of fourth bandpass filters are disposed on a second surface of the support base that is opposite and parallel to the first surface.

15. the beam to be processed by the flow cytometer comprises a beam collected from a microparticle; The optical detection system of claim 2 , wherein the fluorescent signal comprises a fluorescent signal emitted from the microparticle.

16. 1. An optical detection method for a flow cytometer, comprising: a beam separation device separating the beam to be processed by the flow cytometer into a plurality of first beams having distinct wavelength ranges that either do not overlap with one another or partially overlap with one another; a plurality of wavelength division multiplexing devices multiplexing and detecting the plurality of first beams, respectively; Including, each wavelength division multiplexing device comprises a plurality of photodetector devices configured to detect respective ones of the plurality of first beams, the plurality of first beams being parallel to one another when received by the plurality of wavelength division multiplexing devices.

17. the beam to be processed by the flow cytometer comprises a side scatter signal and a fluorescence signal; The optical detection method further includes a side-scattered optical signal separation device separating a side-scattered optical signal from the plurality of first beams; and transmitting the separated plurality of first beams through the wavelength division multiplexing device; (i) the optical detection method further comprises: an attenuation device attenuating the side-scattered light signals separated by the side-scattered light signal separation device; a first bandpass filter performing bandpass filtering on the side-scattered light signal attenuated by the attenuation device; a first light detection unit detecting the side-scattered light signal filtered by the first band-pass filter; Contains, or 17. The optical detection method of claim 16, wherein (ii) the beam to be processed by the flow cytometry method comprises a beam collected from a microparticle, and the fluorescent signal comprises a fluorescent signal emitted from the microparticle.

18. 17. The optical detection method of claim 16, further comprising: a second mirror changing a transmission path of at least one of the plurality of first beams such that the plurality of first beams are transmitted in parallel.

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