Optical detection system and optical detection method for a flow cytometer
Patent Information
- Application Number
- CN202611076503.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2022-03-23
- Publication Date
- 2026-10-13
Smart Images
Figure CN122901054A_ABST
Abstract
Description
[0001] This application is a divisional application of National Application No. 202210288648.8, filed on March 23, 2022, entitled "Optical Detection System and Method for Flow Cytometer". Technical Field
[0002] This disclosure relates to a light detection system and a light detection method for use in flow cytometers. Background Technology
[0003] Optical detection technology has applications in many fields. For example, it can be used to detect the volume, shape, and properties of objects. For instance, it can be used in flow cytometry to detect the volume, shape, and properties of particles, for example. Summary of the Invention
[0004] A brief overview of this disclosure is given below to provide a basic understanding of certain aspects of it. However, it should be understood that this overview is not an exhaustive summary of this disclosure. It is not intended to identify key or essential parts of this disclosure, nor is it intended to limit the scope of this disclosure. Its purpose is merely to present certain concepts of this disclosure in a simplified form as a prelude to the more detailed description that follows.
[0005] According to one aspect of this disclosure, a light detection system for a flow cytometer is provided, comprising: a beam splitter configured to split a light beam to be processed by the flow cytometer into a plurality of first beams, the wavelength ranges of the plurality of first beams not overlapping or partially overlapping each other; and a plurality of wavelength division multiplexing (WDM) devices, each WDM device configured to receive a corresponding first beam from the plurality of first beams. The plurality of first beams are parallel to each other when received by the plurality of WDM devices. Each of the plurality of WDM devices includes a plurality of light detection devices configured to detect portions of the corresponding first beam.
[0006] According to another aspect of this disclosure, a method for optical detection in a flow cytometer is provided, comprising: separating a light beam to be processed by the flow cytometer into a plurality of first beams via a beam splitter, wherein the wavelength ranges of the plurality of first beams do not overlap or partially overlap; and performing wavelength division multiplexing and detection on the plurality of first beams respectively via a plurality of wavelength division multiplexing devices. Each wavelength division multiplexing device includes a plurality of optical detection devices for detecting a corresponding first beam among the plurality of first beams, and the plurality of first beams are parallel to each other when received by the wavelength division multiplexing device.
[0007] In accordance with other aspects of this disclosure, computer program code and computer program products for implementing the methods according to this disclosure are also provided, as well as a computer-readable storage medium having the computer program code for implementing the methods according to this disclosure recorded thereon.
[0008] Other aspects of embodiments of this disclosure are set forth in the following description section, wherein detailed description is provided to fully disclose preferred embodiments of this disclosure, without limiting them. Attached Figure Description
[0009] This disclosure can be better understood by referring to the detailed description given below in conjunction with the accompanying drawings, in which the same or similar reference numerals are used throughout the drawings to denote the same or similar parts. These drawings, together with the following detailed description, are incorporated in and form part of this specification, and are used to further illustrate preferred embodiments of the disclosure and explain the principles and advantages of the disclosure. Wherein:
[0010] Figure 1 This is a block diagram illustrating an example configuration of a light detection system for a flow cytometer according to an embodiment of the present disclosure;
[0011] Figure 2 This is a 3D layout diagram illustrating a specific implementation of a light detection system for a flow cytometer according to an embodiment of the present disclosure;
[0012] Figure 3 This is a top view illustrating a specific implementation of a light detection system for a flow cytometer according to an embodiment of the present disclosure;
[0013] Figure 4 This is a side view illustrating a specific implementation of a light detection system for a flow cytometer according to an embodiment of the present disclosure;
[0014] Figure 5A and Figure 5B This is a diagram illustrating a specific implementation of a beam splitting apparatus according to an embodiment of the present disclosure;
[0015] Figure 6A , Figure 6B and Figure 6C This is a diagram illustrating another specific implementation of the beam splitting apparatus according to an embodiment of the present disclosure;
[0016] Figure 7 This is a top view illustrating another specific implementation of a light detection system for a flow cytometer according to an embodiment of the present disclosure;
[0017] Figure 8 This is a schematic diagram illustrating a specific implementation of a wavelength division multiplexing unit according to an embodiment of the present disclosure;
[0018] Figure 9A An example of a support substrate included in a wavelength division multiplexing apparatus according to an embodiment of the present disclosure is shown;
[0019] Figure 9B This is a schematic diagram showing the arrangement of the wavelength division multiplexing unit on the support substrate according to an embodiment of the present disclosure;
[0020] Figure 10A This is another example illustrating a support substrate included in a wavelength division multiplexing apparatus according to an embodiment of the present disclosure;
[0021] Figures 10B to 10D This is a schematic diagram showing the arrangement of the wavelength division multiplexing unit on the support substrate according to an embodiment of the present disclosure;
[0022] Figure 11 This is a schematic diagram illustrating another specific implementation of the wavelength division multiplexing unit according to an embodiment of the present disclosure;
[0023] Figure 12 This is a schematic diagram illustrating yet another specific implementation of the wavelength division multiplexing unit according to an embodiment of the present disclosure;
[0024] Figure 13 This is a flowchart illustrating an example of a light detection method for a flow cytometer according to embodiments of the present disclosure; and
[0025] Figure 14 This is a block diagram illustrating an example structure of a personal computer that may be employed in embodiments of this disclosure. Detailed Implementation
[0026] Exemplary embodiments of the present disclosure will be described below with reference to the accompanying drawings. For clarity and brevity, not all features of actual implementations are described in the specification. However, it should be understood that many implementation-specific decisions must be made in the development of any such actual embodiment to achieve the developer’s specific goals, such as complying with constraints related to the system and business, and these constraints may vary from implementation to implementation. Furthermore, it should be understood that while development work can be very complex and time-consuming, such development work is merely a routine task for those skilled in the art who benefit from the present disclosure.
[0027] It should be understood that while the terms "first," "second," etc., may be used herein to describe various elements, these elements should not be limited by these terms. These terms are used only to distinguish one element from another. For example, without departing from the scope of this disclosure, a first element may be referred to as a second element, and similarly, a second element may be referred to as a first element.
[0028] Furthermore, in this specification and the accompanying drawings, multiple components with substantially the same functional configuration are distinguished by appending different letters after the same reference numerals. For example, wavelength division multiplexing unit 1046 may be distinguished as wavelength division multiplexing unit 1046a, wavelength division multiplexing unit 1046b, and wavelength division multiplexing unit 1046c as needed. However, when it is not necessary to specifically distinguish each of the multiple components with substantially the same functional configuration, only the same reference numerals are appended. For example, when it is not necessary to specifically distinguish wavelength division multiplexing unit 1046a, wavelength division multiplexing unit 1046b, and wavelength division multiplexing unit 1046c, wavelength division multiplexing unit 1046a, wavelength division multiplexing unit 1046b, and wavelength division multiplexing unit 1046c may be simply referred to as wavelength division multiplexing unit 1046.
[0029] It should also be noted that, in order to avoid obscuring this disclosure with unnecessary details, only the equipment structure and / or processing steps closely related to the solution according to this disclosure are shown in the accompanying drawings, while other details that are not closely related to this disclosure are omitted.
[0030] The embodiments according to this disclosure are described in detail below with reference to the accompanying drawings.
[0031] First, refer to Figures 1 to 7 An implementation example of a light detection system for a flow cytometer according to embodiments of the present disclosure is described. Figure 1 This is a block diagram illustrating a configuration example of a light detection system 100 for a flow cytometer according to an embodiment of the present disclosure. Figure 2 , Figure 3 and Figure 4 These are a 3D layout diagram, a top view, and a side view illustrating a specific implementation of a light detection system 100 for a flow cytometer according to an embodiment of the present disclosure. Figure 5A and Figure 5B This is a diagram illustrating a specific implementation of a beam splitting apparatus according to an embodiment of the present disclosure. Figure 6A , Figure 6B and Figure 6C This is a diagram illustrating another specific implementation of the beam splitting apparatus according to an embodiment of the present disclosure. Figure 7 This is a top view illustrating another specific implementation of a light detection system for a flow cytometer according to an embodiment of the present disclosure.
[0032] like Figures 1 to 4 As shown, the optical detection system 100 for a flow cytometer according to an embodiment of the present disclosure may include a beam splitter 102 and a plurality of wavelength division multiplexing devices 104.
[0033] The beam splitter (also referred to as a "spectral segmentation device") 102 can be configured to split a beam to be processed by a flow cytometer (hereinafter simply referred to as the "beam to be processed") into a plurality of first beams, the wavelength ranges of which do not overlap or partially overlap with each other. For example, the beam to be processed can be transmitted to the beam splitter 102 via an optical fiber. Alternatively, for example, the beam to be processed can be transmitted to the beam splitter 102 via free space using a coupling lens system.
[0034] As an example, the beam splitter 102 may include a long-pass filter or a short-pass filter. For example, such as Figure 2 and Figure 4 As shown, the beam splitter 102 can be placed at a 45° angle to the optical axis of the beam to be processed, splitting the beam into two first beams that propagate along the x-axis and z-axis directions, respectively. For example, when the wavelength range of the beam to be processed is 350 nm to 950 nm, the beam splitter 102 can separate the beam into two first beams with wavelength ranges of 350 nm to A and A to 950 nm, respectively, for example, 400 nm ≤ A ≤ 900 nm. Note that in Figure 4 In the diagram, the element enclosed in a dashed box can correspond to one of the two first beams.
[0035] As another example, the beam splitting device 102 may include a dichroic mirror. For example, such as Figure 5A and Figure 5B As shown, this dichroic beam splitter can reflect a portion of visible light (Vis) and infrared light (IR), while transmitting ultraviolet light (UV) and the remainder of visible light, thereby separating the beam to be processed into two first beams. Furthermore, by arranging two or more dichroic beam splitters with different optical properties, the beam to be processed can be separated into three or more first beams. For example, as... Figure 6A As shown, by arranging them respectively, they have the following characteristics: Figure 6B and Figure 6C The two dichroic beam splitters with different optical properties shown can separate the beam to be processed into three first beams.
[0036] exist Figure 2 In the image, gray lines represent beams of light. Additionally, in... Figure 5A and Figure 6A In the figures, arrows represent light beams. As those skilled in the art will understand, the actual shape of the light beams may differ from that shown in the figures. Furthermore, the relative positions of the first light beams are not limited by the figures.
[0037] Each wavelength division multiplexing device 104 can be configured to receive a corresponding first beam from the plurality of first beams. For example, each wavelength division multiplexing device 104 can correspond one-to-one with a first beam.
[0038] Each wavelength division multiplexing device 104 may include a plurality of optical detection devices 1042 configured to detect (e.g., intensity detection) a portion of a first light beam received by the wavelength division multiplexing device 104. Figure 2 (Not shown). For example, each wavelength division multiplexing device 104 can perform wavelength division multiplexing on the received first beam to obtain multiple second beams, and transmit the multiple second beams to multiple optical detection devices 1042 respectively. Note that although in Figures 2 to 4 The wavelength division multiplexing device 104 shown includes Figure 11 The wavelength division multiplexing unit 1046b shown performs wavelength division multiplexing on the received first beam, but those skilled in the art can use other wavelength division multiplexing units as needed, such as... Figure 8 The wavelength division multiplexing unit 1046a shown Figure 12 The wavelength division multiplexing unit 1046c shown is an example.
[0039] As an example, multiple second beams can be focused onto multiple light detection devices 1042 via multiple second aspherical lenses 122, such as... Figure 3 and Figure 4 As shown.
[0040] The light detection device 1042 can be, for example, an avalanche photodiode (APD).
[0041] Multiple first beams can be parallel to each other when received by multiple wavelength division multiplexing devices 104. In this case, for example, the various optical detection devices 1042 included in the multiple wavelength division multiplexing devices 104 can be arranged on the same plane, thereby facilitating, for example, temperature control of the optical detection devices 1042, and layout of signal processing circuits and wiring harnesses.
[0042] As an example, such as in Figure 4 In the illustrated optical detection system 100a, the transmission path of at least one of a plurality of first beams can be altered via a second reflector 118, so that the plurality of first beams are transmitted in parallel to a plurality of wavelength division multiplexing devices 104. For example, the second reflector 118 can alter the transmission path of a corresponding first beam by performing total internal reflection on the corresponding first beam. The second reflector 118 can be a plane mirror.
[0043] As another example, multiple first beams can be transmitted non-parallel to multiple wavelength division multiplexing devices 104. For example, as in Figure 7In the illustrated optical detection system 100b, two beams can be transmitted to the corresponding wavelength division multiplexing device 104 at a 90° angle to each other. Furthermore, in practical use, the angle between the beams can be changed as needed by configuring the beam separation device 102. For example, the angle between the two beams can be between 15° and 165°.
[0044] Currently, flow cytometry is evolving from traditional multicolor fluorescence channels to high-channel full-spectrum fluorescence. Full-spectrum fluorescence will acquire more fluorescence information, enabling the simultaneous analysis of more luciferin and other information (up to 40 channels or more), thus obtaining more sample expression information and achieving more sensitive and accurate test results. To achieve this goal, high-density channel acquisition of fluorescence spectra is required to reflect the spectral characteristics of fluorescence.
[0045] As described above, in the optical detection system 100 according to an embodiment of the present disclosure, the light beam to be processed (e.g., including a fluorescence signal) can be separated into multiple first beams via a beam splitter 102, and then wavelength division multiplexing and detection can be performed by a wavelength division multiplexing device 104. This allows for easy increases in the number of optical channels (e.g., 20 or more) while maintaining good optical performance. Furthermore, this arrangement of the optical detection system 100 reduces the optical path length and the number of traversed optical elements for the last one or more channels corresponding to the respective first beams compared to the case where wavelength division multiplexing and detection of the light beam to be processed are performed directly by the wavelength division multiplexing device 104. This improves the optical transmission efficiency of the last one or more channels and reduces the size of the wavelength division multiplexing device 104.
[0046] Additionally, the optical detection system 100 is compatible with a two-port wavelength division multiplexing (WDM) structure. Two excitation sources (examples of objects generating the beam to be processed) with a small number of channels (e.g., less than or equal to 10) can share a single two-port WDM structure, thereby saving costs.
[0047] As an example, multiple first beams can be arranged in a vertical direction (e.g., Figure 2 The layered transmission along the z-axis (as shown) allows for, for example, a further reduction in the size of the wavelength division multiplexing device 104. According to embodiments of this disclosure, the light beam to be processed by the flow cytometer may include side-scattered light signals and fluorescence signals. In this case, as... Figure 3 and Figure 4As shown, the optical detection system may further include multiple side-scattered light signal separation devices 106. Each side-scattered light signal separation device 106 may be configured to separate the side-scattered light signal from a corresponding first beam (i.e., the first beam transmitted to the side-scattered light signal separation device 106) among multiple first beams, and transmit the separated corresponding first beam to a corresponding wavelength division multiplexing device (e.g., a wavelength division multiplexing device for wavelength division multiplexing and detection of the separated corresponding first beam), thereby reducing the influence of the side-scattered light signal on the fluorescence signal and improving the signal-to-noise ratio of the fluorescence signal. For example, the side-scattered light signal separation device 106 may correspond one-to-one with the first beam.
[0048] As an example, the beam to be processed may include a beam collected from the particles, and the fluorescence signal may include a fluorescence signal emitted from the particles.
[0049] According to embodiments of this 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. Figure 2 (Not shown). The first side-scattered light signal separation unit 1062 can be configured to separate the side-scattered light signal from the corresponding first beam, and the second side-scattered light signal separation unit 1064 can be configured to further separate the side-scattered light signal from the corresponding first beam after separation by the first side-scattered light signal separation unit 1062, and transmit the separated corresponding first beam to the corresponding wavelength division multiplexing device 104 among the plurality of wavelength division multiplexing devices 104, so that, for example, the signal-to-noise ratio of the fluorescence signal can be further improved.
[0050] For example, the first side-scattered light signal separation unit 1062 may include a long-pass filter. For instance, the long-pass filter may be placed at an angle of 20 to 50° to the optical axis of the corresponding first beam to reflect the side-scattered light signal and allow the fluorescence signal to pass through.
[0051] As an example, the second side-scattered light signal separation unit 1064 can be composed of a long-pass filter. For example, the long-pass filter can be placed at 0±5° with 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.
[0052] For example, such as Figure 3 , Figure 4 and Figure 7As shown, the optical detection system 100 may further include a plurality of attenuation devices 108, a plurality of first bandpass filters 110, and a plurality of first optical detection units 112. Each attenuation device 108 may be configured to attenuate the side-scattered light signal separated by the first side-scattered light signal separation unit 1062 corresponding to the attenuation device 108, to prevent the side-scattered light signal from saturating the first optical detection unit 112. Each first bandpass filter 110 may be configured to bandpass filter the side-scattered light signal attenuated by the attenuation device 108 corresponding to the first bandpass filter 110. Each first optical detection unit 112 may be configured to detect (e.g., intensity detection) the side-scattered light signal filtered by the first bandpass filter 110 corresponding to the first optical detection unit 112, thereby enabling the detection of, for example, the size of particles screened by flow cytometry.
[0053] For example, the attenuation coefficients of the multiple attenuation devices 108 may be the same, or the attenuation coefficient of at least one of the multiple attenuation devices 108 may be different from that of the other attenuation devices 108. Similarly, the multiple first bandpass filters 110 may have the same optical characteristics, or the optical characteristics of at least one of the multiple first bandpass filters 110 may be different from those of the other first bandpass filters 110.
[0054] As an example, when the beam splitter 102 splits the beam to be processed into two first beams, the intensity of the side-scattered light signal included in one first beam (hereinafter referred to as "SSC1") and the intensity of the side-scattered light signal included in the other first beam (hereinafter referred to as "SSC2") can be b% and 1-b%, respectively, of the intensity of the side-scattered light signal included in the beam to be processed (hereinafter referred to as "SSC"), where 1≤b≤2. Side-scattered light signals generated by small particles (such as particles with sizes on the nanometer scale) are relatively weak, so it is desirable to maximize the collection efficiency of side-scattered light signals from small particles. On the other hand, side-scattered light signals generated by large particles (such as cells with sizes on the micrometer scale) are relatively strong and easily saturate the photodetector, so it is desirable to reduce the collection efficiency of side-scattered light signals from large particles. With SSC1 and SSC2 configured as described above, SSC1 can be used to detect cells with sizes ranging from 1 μm to 30 μm, and SSC2 can be used to detect small particles with sizes ranging from 80 nm to 1 μm, thereby further improving the detection accuracy of particles. For example, in Figure 4 In the illustrated optical detection system 100a, the elements outlined by the upper dashed frame and the elements outlined by the lower dashed frame can respectively correspond to the first beam including SSC1 and SSC2. Furthermore, for example, in... Figure 7In the optical detection system 100b shown, the element enclosed by the dashed box with reference numeral SSC1 can be used for the transmission and detection of SSC1, and the element enclosed by the dashed box with reference numeral SSC2 can be used for the transmission and detection of SSC2.
[0055] For example, such as Figures 2 to 4 and Figure 7 As shown, the optical detection system 100 may further include a plurality of first reflectors 114. Each first reflector 114 may be configured to change the transmission path of the side-scattered light signal separated by the first side-scattered light signal separation unit 1062 corresponding to the first reflector 114, so that the side-scattered light signal is transmitted to the corresponding attenuation device 108.
[0056] For example, such as Figure 3 As shown, the first reflector 114 can be placed at an angle of 20° to 50° to the optical axis of the side-scattered light signal from the first side-scattered light signal separation unit 1062 corresponding to the first reflector 114, so as to reflect the side-scattered light signal, thereby changing its transmission path, so that, for example, the side-scattered light signal is transmitted parallel to the first beam separated by the first side-scattered light signal separation unit 1062. In this case, for example, multiple first light detection units 112 can be arranged on the same plane as multiple light detection devices 1042, thereby facilitating, for example, temperature control of the multiple first light detection units 112 and multiple light detection devices 1042, and facilitating the layout of signal processing circuits and wiring harnesses. For example, refer to Figure 3 The virtual reference plane, represented by dashed lines, is used to arrange multiple first optical detection units 112 and multiple optical detection devices 1042, so as to more accurately arrange the multiple first optical detection units 112 and multiple optical detection devices 1042 on the same plane.
[0057] For example, such as Figure 3 , Figure 4 and Figure 7 As shown, the optical detection system 100 may further include a plurality of first aspherical lenses 120 disposed between a plurality of first bandpass filters 110 and a plurality of first optical detection units 112. Each first aspherical lens 120 may be configured to focus a side-scattered light signal filtered by the first bandpass filter 110 corresponding to the first aspherical lens 120 onto the corresponding first optical detection unit 112.
[0058] For example, such as Figure 3 , Figure 4 and Figure 7As shown, the optical detection system 100 may further include an achromatic lens 116 configured to collimate, for example, a beam of light to be processed collected via optical fiber coupling, and to transmit the collimated beam to the beam splitter 102. Furthermore, for example, the beam of light to be processed may be transmitted to the achromatic lens 116 via free space using a coupling lens system.
[0059] The following will refer to Figures 8 to 12 A wavelength division multiplexing unit according to an embodiment of the present disclosure will be described in detail.
[0060] Figure 8 This is a schematic diagram illustrating a specific implementation of a wavelength division multiplexing section according to an embodiment of the present disclosure. For example... Figure 8 As shown, 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 to the plurality of mirrors 1046-M. Each mirror 1046-M may be configured to reflect a light beam transmitted from a corresponding second bandpass filter 1046-L 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., recessed portion) C disposed between two planar portions P. Each concave portion C is configured to reflect a light beam transmitted from a corresponding second bandpass filter 1046-L of the plurality of second bandpass filters 1046-L to the concave portion C to collimate the light beam. In addition, each second bandpass filter 1046-L may be configured to transmit light in the wavelength range corresponding to the second bandpass filter 1046-L and reflect the remaining light. Therefore, the beam propagates in a zigzag pattern between the multiple reflectors 1046-M and the multiple second bandpass filters 1046-L to separate the corresponding beam (e.g., the first beam) into multiple second beams, such as... Figure 8 The diagram shows the 10 second beams corresponding to channels #1 to #10. Furthermore, by providing concave portions C between the planar portions P, the divergent light after transmission over a certain distance is re-collimated to achieve transmission over a longer distance, thereby facilitating the detection of more channels. Note that although in Figure 8 The diagram shows adjacent mirrors 1046-M spaced apart from each other, but multiple mirrors 1046-M can be configured to have no gaps between them as needed. Furthermore, for example, multiple mirrors 1046-M can be configured as a single mirror 1046-M having one or more planar portions P and concave portions C and extending the length of the wavelength division multiplexing section 1046a.
[0061] In some examples, the directivity of the beam can be adjusted by adjusting the relative position between the second bandpass filter 1046-L and the corresponding reflector 1046-M, for example, the distance between the centers of the second bandpass filter 1046-L and the corresponding reflector 1046-M in the direction in which the second bandpass filter 1046-L is arranged, to further ensure that the beam is transmitted in a zigzag pattern.
[0062] As an example, wavelength division multiplexing device 104 may also include, for example, Figure 9A The supporting substrate 1048a is shown. For example, as... Figure 9B As shown, multiple mirrors (also known as "mirror arrays") can be placed on the first surface SA1 of the substrate 1048a, and multiple second bandpass filters (also known as "second bandpass filter arrays") can be placed on the second surface SA2 of the substrate 1048a that is opposite to and parallel to the first surface SA1. This ensures the parallelism between the mirror array and the second bandpass filter array and improves the optical transmission efficiency.
[0063] For example, such as Figure 9B As shown, multiple reflectors 1046-M and multiple second bandpass filters 1046-L can be configured to extend beyond the first surface SA1 and the second surface SA2, respectively. This prevents the light beam from propagating through the interior of the substrate 1048a, thus avoiding interface light loss (Freile reflection loss) caused by light propagating through the interior of the substrate 1048a and further improving light transmission efficiency. Additionally, light scattering caused by crystallization or bubbles within the material of the substrate 1048a can be avoided, thereby reducing crosstalk between different channels.
[0064] As another example, wavelength division multiplexing device 104 may also include, for example, Figure 10A The shown is a support base 1048b with a hollow interior. For example, as... Figures 10B to 10D As shown, multiple reflectors 1046-M can be disposed on the first surface SB1 of the substrate 1048b, and multiple second bandpass filters 1046-L can be disposed on the second surface SB2 of the substrate 1048b, which is opposite to and parallel to the first surface SB1. This ensures the parallelism between the multiple reflectors 1046-M and the multiple second bandpass filters 1046-L, improving optical transmission efficiency. Furthermore, in this configuration, the light beam is transmitted through the hollow interior, thus avoiding interface light loss and further improving optical transmission efficiency. Additionally, light scattering caused by crystallization or bubbles within the material of the substrate 1048b can be avoided, thereby reducing crosstalk between different channels. Moreover, the hollow interior configuration allows for a significant reduction in the mass of the substrate 1048b while maintaining parallelism and structural strength.
[0065] For example, the hollow interior of the support substrate 1048b can be filled with the same material as the support substrate 1048b or any other material with a refractive index greater than 1, such as glass, polymer, fluid, etc.
[0066] For example, two wavelength division multiplexing devices 104 can share a single bearing substrate 1048. For example, in Figure 9B and Figure 10B In the example shown, the upper mirror array and the second bandpass filter array, as well as the lower mirror array and the second bandpass filter array, can each correspond to two different wavelength division multiplexing devices 104, thereby reducing, for example, the size and weight of the wavelength division multiplexing device 104.
[0067] For example, the wavelength division multiplexing unit 1046 included in multiple wavelength division multiplexing devices 104 can be configured as a single unit.
[0068] The support substrate 1048 can be made of transparent or opaque materials. For example, the support substrate 1048 can be formed of metal, glass and / or polymer.
[0069] Figure 11 This is a schematic diagram illustrating another specific implementation of the wavelength division multiplexing unit according to an embodiment of the present disclosure. Figure 11 The wavelength division multiplexing unit 1046b shown is... Figure 8 The difference in the wavelength division multiplexing unit 1046a shown is that the reflector 1046-M is replaced by a plurality of alternately arranged plane mirrors 1046-P and a plurality of concave mirrors 1046-C. Similar to the planar portion P of the reflector 1046-M, each plane mirror 1046-P can be configured to reflect the light beam transmitted from a corresponding second bandpass filter 1046-L among the plurality of second bandpass filters 1046-L to that plane mirror 1046-P. Similarly, similar to the concave portion C of the reflector 1046-M, each concave mirror 1046-C can be configured to reflect the light beam transmitted from the second bandpass filter 1046-L corresponding to that concave mirror among the plurality of second bandpass filters 1046-L to that concave mirror to collimate the light beam. Although a plane mirror 1046-P is shown between two concave mirrors 1046-C in section 11, two or more plane mirrors 1046-P can be arranged between the two concave mirrors 1046-C as needed. For example, as Figure 3 and Figure 7 As shown, two plane mirrors 1046-P are placed between the two concave mirrors 1046-C.
[0070] Figure 12This is a schematic diagram illustrating another specific implementation of a wavelength division multiplexing (WDM) unit according to an embodiment of the present disclosure. The WDM 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 within the wavelength range corresponding to that bandpass filter and reflects the remaining light. For example, as... Figure 12 As shown, a concave mirror can be inserted between some of the third bandpass filters 1046-J among a plurality of third bandpass filters 1046-J to collimate the beam transmitted from the corresponding fourth bandpass filter 1046-K among a plurality of fourth bandpass filters 1046-K to the concave mirror. Compared with wavelength division multiplexing units 1046a and 1046b, wavelength division multiplexing unit 1046c can further increase the number of fluorescence channels. Note that although in Figure 12 The diagram shows only one concave mirror, but two or more concave mirrors can be used as needed. For example, the third bandpass filter 1046-J and the concave mirror can be configured according to reference... Figure 11 The plane mirror 1046-P and the concave mirror 1046-C are arranged in an alternating manner as described. Furthermore, the specific position of the concave mirror is not limited by the accompanying drawings.
[0071] For example, wavelength division multiplexing units 1046b and 1046c can be used with reference to Figures 9A to 10D The wavelength division multiplexing unit 1046a described herein is arranged in a similar manner, and will not be repeated here.
[0072] Regarding the above reference Figures 8 to 12 The wavelength division multiplexing unit 1046 described can be improved by adding corresponding components (e.g., Figure 8 The number of reflectors 1046-M and the second bandpass filter 1046-L shown can be used to easily increase the number of fluorescence channels.
[0073] The above mainly describes the optical detection system used in flow cytometers; however, this optical detection system can also be applied to other devices, such as sorting devices.
[0074] According to embodiments of this disclosure, a light detection method for flow cytometers can be provided. Figure 13 This is a flowchart illustrating a process example of a light detection method 1000 for a flow cytometer according to an embodiment of the present disclosure.
[0075] like Figure 13 As shown, the optical detection method 1000 may include a beam separation step S1020 and a beam detection step S1040.
[0076] In beam separation step S1020, the beam to be processed by the flow cytometer (hereinafter, it may also be referred to as the "beam to be processed") can be separated into multiple first beams via a beam separation device. The wavelength ranges of the multiple first beams do not overlap or partially overlap with each other. For example, the beam to be processed can be separated into multiple first beams via the beam separation device 102 described above.
[0077] In beam detection step S1040, the plurality of first beams can be detected by the plurality of optical detection devices included in the plurality of wavelength division multiplexing devices (e.g., wavelength division multiplexing device 104). For example, the plurality of first beams are parallel to each other when received by the wavelength division multiplexing device.
[0078] In the optical detection method 1000 according to an embodiment of the present disclosure, the light beam to be processed can be separated into a plurality of first beams via a beam splitting device, and then wavelength division multiplexing and detection can be performed by a wavelength division multiplexing device, which makes it easy to increase the number of optical channels (e.g., more than 20) and maintain good optical performance.
[0079] As an example, the beam to be processed may include a side-scattered light signal and a fluorescence signal. In this case, the light detection method 1000 may further include: separating the side-scattered light signal from a plurality of first beams via a side-scattered light signal separation device, and transmitting the separated plurality of first beams to a wavelength division multiplexing device. For example, the scattered light signal may be separated from the plurality of first beams via the side-scattered light signal separation device 106 described above.
[0080] For example, the beam to be processed may include a beam collected from the particles, and the fluorescence signal may include a fluorescence signal emitted from the particles.
[0081] For example, the light detection method 1000 may further include: via an attenuation device (e.g., Figures 2 to 4 The attenuation device 108 shown attenuates the scattered light signal separated by the side-scattered light signal separation device; the signal is then attenuated by the first bandpass filter (e.g., Figures 2 to 4 The first bandpass filter 110 shown performs bandpass filtering on the side-scattered light signal attenuated by the attenuation device, and passes it through the first optical detection unit (e.g., Figure 3 and Figure 4 The first light detection unit 112 shown detects the side-scattered light signal filtered by the first bandpass filter, thereby detecting, for example, the size of the object that is the source of the side-scattered light signal.
[0082] As an example, the light detection method 1000 may further include: changing the transmission path of at least one of the plurality of first beams via a second reflector so that the plurality of first beams are transmitted in parallel.
[0083] It should be noted that although the functional configuration and operation of the optical detection system and optical detection method according to the embodiments of the present disclosure have been described above, these are merely examples and not limitations. Those skilled in the art can modify the above embodiments based on the principles of the present disclosure, such as adding, deleting or combining functional modules and operations in each embodiment, and all such modifications fall within the scope of the present disclosure.
[0084] Furthermore, it should be noted that the details described in the system embodiments can be applied to the method embodiments, and similarly, the details described in the method embodiments can be applied to the system embodiments.
[0085] Furthermore, this disclosure also provides a storage medium and a program product. It should be understood that the machine-executable instructions in the storage medium and program product according to embodiments of this disclosure can also be configured to perform the aforementioned light detection method; therefore, details not described in detail here can be referred to the descriptions in the preceding corresponding sections and will not be repeated here.
[0086] Accordingly, the storage medium used to carry the aforementioned program product including machine-executable instructions is also included in the disclosure of this invention. This storage medium includes, but is not limited to, floppy disks, optical disks, magneto-optical disks, memory cards, memory sticks, etc.
[0087] Furthermore, it should be noted that the aforementioned series of processes and devices can also be implemented via software and / or firmware. In the case of software and / or firmware implementation, data can be transferred from storage media or networks to computers with dedicated hardware architectures, such as… Figure 14 The general-purpose personal computer 1700 shown is equipped with the programs that constitute this software, and when various programs are installed, the computer is able to perform various functions, etc.
[0088] exist Figure 14 In this system, the central processing unit (CPU) 1701 performs various processes based on the program stored in the read-only memory (ROM) 1702 or the program loaded into the random access memory (RAM) 1703 from the storage section 1708. The RAM 1703 also stores, as needed, the data required when the CPU 1701 performs various processes.
[0089] CPU 1701, ROM 1702 and RAM 1703 are connected to each other via bus 1704. Input / output interface 1705 is also connected to bus 1704.
[0090] The following components are connected to the input / output interface 1705: input section 1706, including a keyboard, mouse, etc.; output section 1707, including a display, such as a cathode ray tube (CRT), liquid crystal display (LCD), etc., and speakers, etc.; storage section 1708, including a hard disk, etc.; and communication section 1709, including a network interface card, such as a LAN card, modem, etc. The communication section 1709 performs communication processing via a network, such as the Internet.
[0091] As needed, drive 1710 is also connected to input / output interface 1705. Removable media 1711, such as disks, optical disks, magneto-optical disks, semiconductor memories, etc., are installed on drive 1710 as needed, so that computer programs read from them can be installed into storage section 1708 as needed.
[0092] When the above series of processes are implemented through software, the program constituting the software is installed from a network such as the Internet or a storage medium such as removable media 1711.
[0093] Those skilled in the art will understand that such storage media are not limited to Figure 14 The illustration shows a removable medium 1711 containing a program, distributed separately from the device to provide the program to the user. Examples of removable media 1711 include magnetic disks (including floppy disks (registered trademark)), optical disks (including optical disc read-only memory (CD-ROM) and digital versatile disks (DVD)), magneto-optical disks (including mini-discs (MD) (registered trademark)), and semiconductor memory. Alternatively, the storage medium may be ROM 1702, a hard disk included in storage section 1708, etc., containing programs and distributed to the user along with the device containing them.
[0094] Preferred embodiments of the present disclosure have been described above with reference to the accompanying drawings, but the present disclosure is by no means limited to the examples described above. Various changes and modifications can be made by those skilled in the art within the scope of the appended claims, and it should be understood that such changes and modifications naturally fall within the technical scope of the present disclosure.
[0095] For example, the multiple functions included in one unit in the above embodiments can be implemented by separate devices. Alternatively, the multiple functions implemented by multiple units in the above embodiments can be implemented by separate devices respectively. In addition, one of the above functions can be implemented by multiple units. Needless to say, such a configuration is included within the scope of the present disclosure.
[0096] In this specification, the steps described in the flowchart include not only processes executed sequentially in the stated order, but also processes executed in parallel or individually, rather than necessarily sequentially. Furthermore, even within the steps of sequential processing, needless to say, the order can be appropriately altered.
[0097] For example, this disclosure may also include the following technical solutions:
[0098] 1. A light detection system for flow cytometers, comprising:
[0099] A beam splitter is configured to split a beam to be processed by the flow cytometer into a plurality of first beams, the wavelength ranges of which do not overlap or partially overlap; and
[0100] Multiple wavelength division multiplexing (WDM) devices are configured to receive a corresponding first beam from a plurality of first beams, the plurality of first beams being parallel to each other when received by the plurality of WDM devices.
[0101] Each of the plurality of wavelength division multiplexing devices includes a plurality of optical detection devices configured to detect portions of the corresponding first beam.
[0102] 2. The light detection system according to claim 1, wherein the light beam to be processed by the flow cytometer includes a side-scattered light signal and a fluorescence signal, and
[0103] The optical detection system further includes multiple side-scattered light signal separation devices, each of which is configured to separate the side-scattered light signal from a corresponding first beam among the multiple first beams, and to transmit the separated corresponding first beam to a corresponding wavelength division multiplexing device among the multiple wavelength division multiplexing devices.
[0104] 3. The optical detection system according to claim 2, wherein each side-scattered light signal separation device comprises:
[0105] The first side-scattered light signal separation unit is configured to separate the side-scattered light signal from the corresponding first beam;
[0106] The second side-scattered light signal separation unit is configured to further separate the side-scattered light signal from the separated corresponding first beam and transmit the separated corresponding first beam to the corresponding wavelength division multiplexing device among the plurality of wavelength division multiplexing devices.
[0107] 4. The optical detection system according to claim 3 further includes:
[0108] Multiple attenuation devices, each configured to attenuate the side-scattered light signal separated by the first side-scattered light signal separation unit included in the corresponding side-scattered light signal separation device;
[0109] A plurality of first bandpass filters, each configured to bandpass filter the side-scattered light signal attenuated by a corresponding attenuation device among the plurality of attenuation devices, and
[0110] Multiple first optical detection units, each configured to detect side-scattered light signals filtered by a corresponding first bandpass filter among the multiple first bandpass filters.
[0111] 5. The optical detection system according to claim 4 further includes:
[0112] A plurality of first reflectors, each first reflector being configured to alter the transmission path of the side-scattered light signal separated by the first side-scattered light signal separation unit included in the corresponding side-scattered light signal separation device, so that the side-scattered light signal is transmitted to a corresponding attenuation device.
[0113] 6. The light detection system according to claim 5, wherein separating the light beam to be processed by the flow cytometer into a plurality of first light beams includes separating the light beam to be processed by the flow cytometer into two first light beams, and
[0114] Wherein, the intensity of the side-scattered light signal included in one of the two first beams and the intensity of the side-scattered light signal included in the other of the two first beams are respectively b% and 1-b% of the intensity of the side-scattered light signal included in the beam to be processed by the flow cytometer, where 1≤b≤2.
[0115] 7. The optical detection system according to claim 4 further includes a plurality of first aspherical lenses disposed between the plurality of first bandpass filters and the plurality of first optical detection units, each first aspherical lens being configured to focus a side-scattered light signal filtered by a corresponding first bandpass filter among the plurality of first bandpass filters onto a corresponding first optical detection unit among the plurality of first optical detection units.
[0116] 8. The optical detection system according to claim 1 further includes:
[0117] An achromatic lens is configured to collimate the light beam to be processed by the flow cytometer and to transmit the collimated light beam to the beam splitter.
[0118] 9. The optical detection system according to claim 1 further includes:
[0119] The second reflector is configured to alter the transmission path of at least one of the plurality of first beams, such that the plurality of first beams are transmitted in parallel to the plurality of wavelength division multiplexing devices.
[0120] 10. The optical detection system according to claim 1 further includes a plurality of second aspherical lenses configured to focus a plurality of second beams of the respective first beam separated by a respective wavelength division multiplexing device onto the plurality of optical detection devices in the respective wavelength division multiplexing device.
[0121] 11. The optical detection system according to claim 1, wherein the beam separation device comprises a long-pass filter or a short-pass filter.
[0122] 12. The optical detection system according to claim 1, wherein each wavelength division multiplexing device includes a wavelength division multiplexing section, the wavelength division multiplexing section including a plurality of mirrors and a plurality of second bandpass filters disposed opposite to the plurality of mirrors.
[0123] Each mirror is configured to reflect the light beam transmitted to it from a corresponding second bandpass filter among the plurality of second bandpass filters.
[0124] 13. The light detection system according to claim 12, wherein each reflector includes a concave portion disposed between two planar portions,
[0125] Each concave portion is configured to reflect a beam of light transmitted from a corresponding second bandpass filter among the plurality of second bandpass filters to the concave portion in order to collimate the beam.
[0126] 14. The light detection system according to 13, wherein the plurality of reflectors are integrally arranged.
[0127] 15. The optical detection system according to claim 1, wherein each wavelength division multiplexing device includes a wavelength division multiplexing section, the wavelength division multiplexing section including a plurality of plane mirrors and a plurality of concave mirrors arranged alternately, and a plurality of second bandpass filters arranged opposite to the plurality of plane mirrors and the plurality of concave mirrors.
[0128] Each plane mirror is configured to reflect the light beam transmitted to the plane portion from a corresponding second bandpass filter among the plurality of second bandpass filters.
[0129] Each concave mirror is configured to reflect a light beam transmitted from a corresponding second bandpass filter of the plurality of second bandpass filters to the concave portion in order to collimate the light beam.
[0130] 16. The optical detection system according to 12, wherein each wavelength division multiplexing device further includes a support substrate, the plurality of mirrors are disposed on a first surface of the support substrate, and the plurality of second bandpass filters are disposed on a second surface of the support substrate that is opposite to and parallel to the first surface.
[0131] 17. The optical detection system according to 16, wherein the supporting substrate has a hollow interior.
[0132] 18. The optical detection system according to claim 1, wherein the plurality of optical detection devices are arranged on the same plane.
[0133] 19. The optical detection system according to claim 1, wherein each wavelength division multiplexing device includes a wavelength division multiplexing unit, the wavelength division multiplexing unit including a plurality of third bandpass filters and a plurality of fourth bandpass filters disposed opposite to the plurality of third bandpass filters.
[0134] 20. The optical detection system according to 19, wherein the wavelength division multiplexing device further includes a support substrate having a hollow interior, the plurality of third bandpass filters are disposed on a first surface of the support substrate, and the plurality of fourth bandpass filters are disposed on a second surface of the support substrate that is opposite to and parallel to the first surface.
[0135] 21. The light detection system according to claim 2, wherein the light beam to be processed by the flow cytometer comprises a light beam collected from microparticles, and
[0136] The fluorescence signal includes the fluorescence signal emitted from the particles.
[0137] 22. A light detection method for flow cytometry, comprising:
[0138] The light beam to be processed by the flow cytometer is separated into multiple first beams via a beam splitter, wherein the wavelength ranges of the multiple first beams do not overlap or partially overlap; and
[0139] The plurality of first beams are wavelength divided and detected by multiple wavelength division multiplexing devices, wherein each wavelength division multiplexing device includes multiple optical detection devices for detecting a corresponding first beam among the plurality of first beams, and
[0140] The plurality of first beams are parallel to each other when received by the plurality of wavelength division multiplexing devices.
[0141] 23. The light detection method according to 22, wherein the light beam to be processed by the flow cytometer includes a side-scattered light signal and a fluorescence signal.
[0142] The optical detection method further includes: separating the side-scattered light signal from the plurality of first beams via a side-scattered light signal separation device, and transmitting the separated plurality of first beams to the wavelength division multiplexing device.
[0143] 24. The optical detection method according to 23 further includes:
[0144] The side-scattered light signal separated by the side-scattered light signal separation device is attenuated by an attenuation device;
[0145] The side-scattered light signal attenuated by the attenuation device is band-pass filtered by the first band-pass filter, and
[0146] The side-scattered light signal filtered by the first bandpass filter is detected by the first optical detection unit.
[0147] 25. The optical detection method according to 22 further includes:
[0148] The transmission path of at least one of the plurality of first beams is altered via a second reflector so that the plurality of first beams are transmitted in parallel.
[0149] 26. The light detection method according to 23, wherein the light beam to be processed by the flow cytometer comprises a light beam collected from microparticles.
[0150] The fluorescence signal includes the fluorescence signal emitted from the particles.
Claims
1. A light detection system (100) for a flow cytometer, comprising: The beam splitting device (102) is configured to split the beam into a plurality of first beams having respective wavelength ranges that do not overlap or partially overlap with each other; as well as Multiple wavelength division multiplexing devices (104), each configured to receive a corresponding first beam from the plurality of first beams. Each of the plurality of wavelength division multiplexing devices (104) includes: Multiple optical detection devices (1042) are configured to detect portions of the first beam received by the wavelength division multiplexing device (104); and The wavelength division multiplexing unit (1046c) is configured to perform wavelength division multiplexing on the first beam received by the wavelength division multiplexing device (104). The wavelength division multiplexing unit (1046c) includes: Multiple first bandpass filters (1046-J); and A plurality of second bandpass filters (1046-K) are disposed opposite to the plurality of first bandpass filters (1046-J). Each of the first bandpass filter (1046-J) and the second bandpass filter (1046-K) is configured to transmit light having wavelengths within the wavelength range corresponding to the bandpass filter and reflect other light. The first bandpass filter (1046-J) and the second bandpass filter (1046-K) are arranged such that light reflected from one of the first bandpass filter (1046-J) and the second bandpass filter (1046-K) is incident on the other of the first bandpass filter (1046-J) and the second bandpass filter (1046-K), thereby causing the first beam to propagate between the first bandpass filter (1046-J) and the second bandpass filter (1046-K) and be separated into a plurality of second beams corresponding to the respective wavelength channels.
2. The optical detection system according to claim 1, wherein, The first beam propagates between the plurality of first bandpass filters (1046-J) and the plurality of second bandpass filters (1046-K) through repeated reflections.
3. The optical detection system according to claim 1 or 2, wherein, The wavelength division multiplexing unit (1046c) further includes at least one concave mirror disposed between adjacent first bandpass filters in the plurality of first bandpass filters (1046-J).
4. The optical detection system according to claim 3, wherein, The at least one concave mirror is configured to collimate the light beam transmitted from a corresponding second bandpass filter (1046-K) of the plurality of second bandpass filters (1046-K) to the concave mirror.
5. The optical detection system according to claim 3 or 4, wherein, Each of the at least one concave mirror includes a concave portion (C) disposed between a pair of planar portions (P), the planar portions (P) being configured to reflect light beams transmitted from a corresponding second bandpass filter among the plurality of second bandpass filters (1046-K), and the concave portion (C) being configured to reflect and collimate light beams transmitted from a corresponding second bandpass filter among the plurality of second bandpass filters (1046-K).
6. The optical detection system according to any one of claims 3 to 5, wherein, The at least one concave mirror is arranged to maintain the propagation of the first beam along a defined optical path between the plurality of first bandpass filters (1046-J) and the plurality of second bandpass filters (1046-K).
7. The optical detection system according to any one of claims 1 to 6, wherein, At least a portion of the light transmitted by the first bandpass filter (1046-J) and the second bandpass filter (1046-K) is received by the plurality of light detection devices (1042).
8. The optical detection system according to any one of claims 1 to 7, wherein, The plurality of first beams are parallel to each other when received by the plurality of wavelength division multiplexing devices (104).
9. The optical detection system according to any one of claims 1 to 8 further includes a second reflector configured to alter the transmission path of at least one of the plurality of first beams such that the plurality of first beams are parallel when received by the plurality of wavelength division multiplexing devices (104).
10. A light detection method for flow cytometry, comprising: The beam is split into multiple first beams having corresponding wavelength ranges that do not overlap or partially overlap with each other; Supplying a corresponding first beam from the plurality of first beams to a plurality of wavelength division multiplexing devices (104); and Each first beam is wavelength divided multiplexed using a wavelength division multiplexing unit (1046c). The wavelength division multiplexing unit (1046c) includes a plurality of first bandpass filters (1046-J) and a plurality of second bandpass filters (1046-K) disposed opposite to the plurality of first bandpass filters (1046-J). Each of the first bandpass filter (1046-J) and the second bandpass filter (1046-K) transmits light having wavelengths within the wavelength range corresponding to the bandpass filter and reflects other light. In this process, light reflected from one of the first bandpass filter (1046-J) and the second bandpass filter (1046-K) is incident on the other of the first bandpass filter (1046-J) and the second bandpass filter (1046-K), such that the first beam propagates between the bandpass filters and is split into a plurality of second beams corresponding to the respective wavelength channels.
11. The method according to claim 10, wherein, The first beam propagates between the bandpass filters through repeated reflections.
12. The method according to claim 10 or 11, further comprising using at least one concave mirror disposed between adjacent first bandpass filters in the plurality of first bandpass filters (1046-J) to collimate the first beam.
13. The method according to claim 12, wherein, The concave mirror collimates the light beam transmitted from the corresponding second bandpass filter (1046-K) to the concave mirror.
14. The method according to any one of claims 12 or 13, wherein, The concave mirror includes a concave portion (C) disposed between a pair of planar portions (P), the planar portions (P) reflecting light beams transmitted from a corresponding second bandpass filter (1046-K), and the concave portion (C) reflecting and collimating light beams transmitted from a corresponding second bandpass filter (1046-K).
15. The method according to any one of claims 10 to 14, further comprising using a plurality of optical detection devices (1042) to detect the plurality of second beams.