A reflective fluorescence collection device for a flow cytometer

Abstract

The present invention discloses a reflective fluorescence collection device for a flow cytometer, which consists of an objective lens group and a spectroscopic component composed of a plano-convex reflective lens and a meniscus aspherical lens, and a forward collection component composed of an aspherical lens. The fluorescence collected by the objective lens group and the spectroscopic component is detected by a spectroscopic module composed of a spectroscope, a focusing lens, and a detector. The present invention uses a reflective fluorescence receiving structure, and only two lenses are used to achieve a numerical aperture of NA 1.2-1.3, and at the same time solves the problem of spatial separation of multiple lasers used in the flow cytometer.

Description

Technical Field

[0001] The present invention relates to the technical field of optical instrument analysis, and particularly to a reflective fluorescence collection device for a flow cytometer. Background Art

[0002] Currently, a flow cytometer is an instrument for analyzing and sorting single-file cells or particles. The cell diversion focusing module can align the cells in a row and keep them advancing in the center of the axis through hydrodynamic or acoustic focusing technology. A flow cytometer usually uses a laser as the light source, and the excitation light source irradiates the cells passing through the diversion cavity in the vertical direction. When the cells pass through, forward scatter and side scatter are generated when the light beam passes through the cells.

[0003] Currently, due to the influence of the wall thickness of the flow cell, the numerical aperture of the microscope objective used for fluorescence collection in a flow cytometer is difficult to reach above 1.0, and the collected fluorescence signal is limited. Since a flow cytometer usually requires multiple excitation light sources to detect multiple fluorescences through spatial separation of multiple lasers, the fluorescence signals emitted by different light sources in the existing optical collection device cannot be dispersed, resulting in interference between different light sources. Summary of the Invention

[0004] The purpose of the present invention is to provide a reflective fluorescence collection device for a flow cytometer, aiming to solve the technical problem that in the existing technology, the fluorescence signals emitted by different light sources in the existing optical collection device cannot be dispersed, resulting in interference between different light sources.

[0005] To achieve the above purpose, a reflective fluorescence collection device for a flow cytometer adopted by the present invention includes a flow cell for the test particle diversion focusing module for fluorescence labeling, an objective lens group for collecting the side scatter and fluorescence excited by the laser irradiating the central particle in the flow cell, a beam splitting component, and a forward collection component;

[0006] The objective lens group is fixedly glued to the outer wall of the flow cell. The numerical aperture of the objective lens group is 1.2 - 1.3, the field of view range is 500μm × 500μm, the wavelength range is 400 - 800nm, and the magnification is 7 - 14. The forward collection component is used to collect forward scatter.

[0007] The beam splitting component includes a fluorescence collimating lens, a first beam splitter, a second beam splitter, a third beam splitter, a fourth beam splitter, and a fifth beam splitter. The fluorescence collimating lens is used to collimate the fluorescence collected by the objective lens group, and is sequentially split by the first beam splitter, the second beam splitter, the third beam splitter, the fourth beam splitter, and the fifth beam splitter, and then the light passing focusing lens focuses the split light onto the forward collection device.

[0008] Among them, the first beam splitter, the second beam splitter, the third beam splitter, the fourth beam splitter, and the fifth beam splitter have different wavelengths.

[0009] Among them, the objective lens assembly includes a first lens and a second lens. The first lens and the second lens are arranged coaxially in sequence. The plane of the first lens is fixed by optical glue to the wide side of the flow cell, and the concave surface of the second lens faces the plane of the flow cell.

[0010] Among them, the first lens is a spherical mirror with a positive optical power, and the material is crown glass with a low refractive index; the second lens is a meniscus aspherical lens with a combination of a positive optical power aspherical surface and a negative optical power, and the material is crown glass with a high refractive index.

[0011] Among them, the forward collection component includes a focusing lens and a detector. The focusing lens is used to collect forward scattered light and transmit it to the detector for detection.

[0012] Among them, the first beam splitter, the second beam splitter, the third beam splitter, the fourth beam splitter, and the fifth beam splitter have a certain angle with the collimated light of the collimating lens, and the angle range is between 10° and 20°.

[0013] Among them, the reflective fluorescence collection device for a flow cytometer further includes an aspherical lens and a photodiode. The flow cell emits scattered light, and the aspherical lens is used to focus the scattered light generated by the flow cell onto the photodiode.

[0014] A reflective fluorescence collection device for a flow cytometer according to the present invention includes the objective lens group composed of a plano-convex reflective lens and a meniscus aspherical lens and the beam splitting component, and the forward collection component composed of an aspherical lens. The fluorescence collected by the objective lens group and the beam splitting component is detected by a beam splitting module composed of a beam splitter, a focusing lens, and a detector. The present invention uses a reflective fluorescence receiving structure, and only two lenses are used to achieve a numerical aperture of NA 1.2 - 1.3, and at the same time solves the problem of spatial separation of multiple lasers used in a flow cytometer. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0016] Figure 1 It is the optical path structure diagram of the reflective fluorescence collection device for a flow cytometer of the present invention.

[0017] Figure 2 This is the first field of view effect diagram of the present invention.

[0018] Figure 3 This is the second field of view effect diagram of the present invention.

[0019] Figure 4 This is the third field of view effect diagram of the present invention.

[0020] In the figure: 1 - first lens, 2 - flow cell, 3 - second lens, 4 - aspherical lens, 5 - photodiode, 6 - fluorescence collimating lens, 7 - first beam splitter, 8 - second beam splitter, 9 - third beam splitter, 10 - fourth beam splitter, 11 - fifth beam splitter, 12 - detector, 13 - light passing focusing lens. Detailed implementation manners

[0021] The embodiments of the present invention will be described in detail below. The examples of the embodiments are shown in the accompanying drawings, where the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below by referring to the drawings are exemplary and are intended to explain the present invention, and should not be construed as limiting the present invention.

[0022] In the description of the present invention, it should be understood that the orientation or positional relationship indicated by the terms "length", "width", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. is based on the orientation or positional relationship shown in the accompanying drawings, and is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention. In addition, in the description of the present invention, the meaning of "plurality" is two or more unless otherwise specifically defined.

[0023] Please refer to Figure 1 , the present invention provides a reflective fluorescence collection device for a flow cytometer, including a test particle diversion focusing module flow cell 2 for fluorescence labeling, an objective lens group for collecting the side scattered light and fluorescence excited by the laser irradiated on the central particle of the flow cell 2, a beam splitting component, and a forward collection component;

[0024] The objective lens group is fixedly glued to the outer wall of the flow cell 2. The numerical aperture of the objective lens group is 1.2 - 1.3, the field of view range is 500μm × 500μm, the wavelength range is 400 - 800nm, the magnification is 7 - 14, and the forward collection component is used to collect forward scattered light;

[0025] The beam splitting component includes a fluorescence collimating lens 6, a first beam splitter 7, a second beam splitter 8, a third beam splitter 9, a fourth beam splitter 10, and a fifth beam splitter 11. The fluorescence collimating lens 6 is used to collimate the fluorescence collected by the objective lens group, and the light is sequentially split by the first beam splitter 7, the second beam splitter 8, the third beam splitter 9, the fourth beam splitter 10, and the fifth beam splitter 11, and then the light passing focusing lens 13 focuses the split light onto the forward collection device.

[0026] Further, the wavelengths of the first beam splitter 7, the second beam splitter 8, the third beam splitter 9, the fourth beam splitter 10, and the fifth beam splitter 11 are different.

[0027] Further, please refer to Figure 1 , the objective lens assembly includes a first lens 1 and a second lens 3. The first lens 1 and the second lens 3 are arranged coaxially in sequence, and the plane of the first lens 1 is fixedly bonded to the wide side of the flow cell 2 through optical glue, and the concave surface of the second lens 3 faces the plane of the flow cell 2.

[0028] Further, the first lens 1 is a spherical mirror with a positive optical power, and the material is crown glass with a low refractive index; the second lens 3 is a meniscus aspherical lens with a combination of a positive optical power aspherical surface and a negative optical power, and the material is crown glass with a high refractive index.

[0029] Further, please refer to Figure 1 , the forward collection assembly includes a focusing lens and a detector 12. The focusing lens is used to collect forward scattered light and transmit it to the detector 12 for detection.

[0030] Further, the first beam splitter 7, the second beam splitter 8, the third beam splitter 9, the fourth beam splitter 10, and the fifth beam splitter 11 have a certain angle with the collimated light of the collimating lens, and the angle range is between 10° and 20°.

[0031] Further, please refer to Figure 1 , the reflection type fluorescence collection device for a flow cytometer further includes an aspherical lens 4 and a photodiode 5. The flow cell 2 emits scattered light, and the aspherical lens 4 is used to focus the scattered light generated by the flow cell 2 onto the photodiode 5.

[0032] In this embodiment, the first lens 1, the second lens 3, and the flow cell 2 of the objective lens assembly are all made of fused quartz glass. The aspect ratio of the inner flow channel is greater than 2.4. The distance from the center of the flow channel to the surface of the first lens 1 is 1.5 - 2 mm, and the distance from the other side to the surface of the second lens 3 is 1.5 - 2 mm. The first lens 1 is a spherical mirror with positive optical power, and the material is crown glass with a low refractive index. The thickness of the first lens 1 is 3 mm. The plane of the first lens 1 is glued and fixed to the flow cell 2. The second lens 3 is a meniscus aspherical lens with a combination of positive optical power aspherical and negative optical power, and the material is crown glass with a high refractive index. The concave surface of the second lens 3 faces the plane of the flow cell 2, and the thickness of the second lens 3 is 3 mm. The numerical aperture of the microscopic objective lens group is 1.2 - 1.3, the field of view range is 500 μm × 500 μm, the wavelength range is 400 - 800 nm, and the magnification is 7 - 14, as Figures 2 to 4 shown; the fluorescence collimating lens 6 collimates the fluorescence collected by the objective lens group, and is sequentially split by the first beam splitter 7, the second beam splitter 8, the third beam splitter 9, the fourth beam splitter 10, and the fifth beam splitter 11, and then focused onto the detector 12 by the focusing lens. Thus, the objective lens group composed of a plano-convex reflecting lens and a meniscus aspherical lens, and the beam splitting assembly, and the forward collection assembly composed of an aspherical lens. The fluorescence collected by the objective lens group and the beam splitting assembly is detected by the beam splitting module composed of a beam splitter, a focusing lens, and the detector 12. The present invention uses a reflective fluorescence receiving structure, and only two lenses are used to achieve a numerical aperture of NA1.2 - 1.3, and at the same time solves the problem of spatial separation of multiple lasers used in flow cytometers.

[0033] The above-disclosed is only a preferred embodiment of the present invention, and of course, it cannot be used to limit the scope of the rights of the present invention. Those of ordinary skill in the art can understand the entire or part of the process of implementing the above embodiment, and the equivalent changes made according to the claims of the present invention still fall within the scope covered by the invention.

Claims

1. A reflective fluorescence collection device for a flow cytometer, characterized in that, It includes a flow cell of a test particle diversion focusing module for fluorescence labeling, an objective lens group for collecting the side-scattered light and fluorescence excited by the laser irradiating the particles at the center of the flow cell, a spectroscopic component, and a forward collection component; The objective lens group is fixedly glued to the outer wall of the flow cell. The numerical aperture of the objective lens group is 1.2 - 1.3, the field of view range is 500μm × 500μm, the wavelength range is 400 - 800nm, and the magnification is 7 - 14. The forward collection component is used to collect the forward scattered light; The spectroscopic component includes a fluorescence collimating lens, a first beam splitter, a second beam splitter, a third beam splitter, a fourth beam splitter, and a fifth beam splitter. The fluorescence collimating lens is used to collimate the fluorescence collected by the objective lens group, and is spectroscopically analyzed successively through the first beam splitter, the second beam splitter, the third beam splitter, the fourth beam splitter, and the fifth beam splitter, and then the spectroscopically analyzed light is focused by a focusing lens onto the forward collection device; The objective lens assembly is composed of a first lens and a second lens. The first lens and the second lens are arranged coaxially in sequence. The first lens and the second lens are respectively arranged on both sides of the flow cell, and the plane of the first lens is fixedly glued to the wide side of the flow cell through optical glue, and the concave surface of the second lens faces the plane of the flow cell; The first lens is a spherical mirror with positive optical power, and the material is crown glass with low refractive index; the second lens is a meniscus aspherical lens with a combination of positive aspherical optical power and negative optical power, and the material is crown glass with high refractive index.

2. The reflective fluorescence collection device for a flow cytometer according to claim 1, wherein, The first beam splitter, the second beam splitter, the third beam splitter, the fourth beam splitter, and the fifth beam splitter have different wavelengths.

3. The reflective fluorescence collection device for a flow cytometer according to claim 1, wherein, The forward collection component includes a focusing lens and a detector. The focusing lens is used to collect the forward scattered light and transmit it to the detector for detection.

4. The reflective fluorescence collection device for a flow cytometer according to claim 1, wherein, The first beam splitter, the second beam splitter, the third beam splitter, the fourth beam splitter, and the fifth beam splitter have a certain angle with the collimated light of the collimating lens, and the angle range is between 10° and 20°.

5. The reflective fluorescence collection device for a flow cytometer according to claim 1, wherein, The reflective fluorescence collection device for a flow cytometer further includes an aspherical lens and a photodiode. The flow cell emits scattered light, and the aspherical lens is used to focus the scattered light generated by the flow cell onto the photodiode.

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