Microfluidic flow cytometry fluorescence detector

Abstract

The utility model relates to a micro-fluidic flow cytometry fluorescence detector which comprises an excitation light source optical fiber, a light beam shaping module, a focusing and fluorescence collecting module and a micro-fluidic chip which are sequentially arranged along the excitation light path direction of the excitation light source optical fiber, and a multi-channel silicon photomultiplier tube is arranged above the micro-fluidic chip. A plurality of groups of detection channels for receiving fluorescence signals of different wave bands are arranged in the multichannel silicon photomultiplier tube at intervals up and down, and a dichroscope, a band-pass filter and a lens group are sequentially arranged in each group of detection channel along a light path and are gathered on a receiving chip; the micro-fluidic flow cytometry fluorescence detector can efficiently and accurately complete multi-wavelength fluorescence excitation and signal collection, and is suitable for multi-channel and multi-wavelength fluorescence detection requirements in a micro-fluidic chip.

Description

TECHNICAL FIELD

[0001] The utility model relates to a micro -fluidic flow cytometer fluorescence detector. BACKGROUND

[0002] With the rapid development of microfluidic technology, the application of microfluidic chips in the fields of life science, medical diagnosis, environmental monitoring, etc. gradually increases. The advantage of microfluidic technology is that it can perform precise liquid control in a small space, enabling a large number of experiments to be performed in a very small volume, thereby achieving rapid and efficient analysis. In microfluidic flow cytometry, fluorescence detection technology has become an important means, which can realize high-sensitivity detection of cell or molecular markers through excitation and emission of fluorescent markers, and is widely used in cytology, immunology, genomics, etc. Research field.

[0003] In microfluidic flow cytometry, excitation and collection of fluorescence signals are crucial. In order to ensure accurate collection and effective transmission of fluorescence signals to detection equipment, precise optical systems are usually required. In traditional microfluidic fluorescence detection devices, excitation light sources are usually collimated and focused by optical elements such as lenses, mirrors, filters, etc. Finally, the sample in the flow channel is excited to produce fluorescence signals. Fluorescence signals are collected by optical fibers or other sensing devices and transmitted to subsequent detection modules for analysis.

[0004] However, the existing microfluidic fluorescence detection device often has a series of problems in application. First, in the existing technology, the transmission path of the signal is complex during excitation and reception, and the light loss is large, especially for multi-wavelength signal processing, multiple dichroic mirrors, filters, etc. Elements such as elements are often used in the system for wavelength separation and filtering. These elements not only increase the volume of the system, but also reduce the overall light collection efficiency, affecting the sensitivity and accuracy of the detection. Secondly, the existing objective lens design is mostly focused on single-wavelength excitation and reception. For complex detection tasks that require multi-wavelength excitation, traditional design often cannot efficiently process multi-wavelength signals, resulting in signal cross interference during multi-wavelength excitation and fluorescence signal reception, reducing the accuracy and efficiency of detection. In addition, many existing fluorescence detection systems rely on complex optical path design, and the arrangement and combination of multiple optical elements increase the overall volume of the system, and the adjustment and calibration are difficult. For space-limited application scenarios such as microfluidic chips, traditional systems are usually difficult to meet the needs of high integration and compact design.

[0005] In order to overcome these problems in the prior art, there is an urgent need for a new microfluidic flow cytometry fluorescence detection module that can simplify the optical path design, integrate the excitation light source and the fluorescence signal receiving system, and optimize the efficiency of multi-wavelength excitation and reception. This new module can realize simultaneous excitation and efficient collection of multi-wavelength fluorescence signals, while providing a compact and highly integrated design to meet the needs of modern microfluidic chip fluorescence detection. Utility model content

[0006] In view of the deficiencies of the prior art, the technical problem to be solved by the utility model is to provide a microfluidic flow cytometry fluorescence detector, which aims to solve the problems of many optical elements, large optical loss, and large system volume in traditional microfluidic fluorescence detection systems through integrated design. The module can efficiently and accurately complete multi-wavelength fluorescence excitation and signal collection, and is suitable for multi-channel, multi-wavelength fluorescence detection requirements in microfluidic chips.

[0007] In order to solve the above technical problems, the technical scheme of the utility model is: a microfluidic flow cytometry fluorescence detector, comprising an excitation light source optical fiber, and a beam shaping module, a focusing and fluorescence collection module, and a microfluidic chip are sequentially arranged along the excitation light source optical fiber excitation light path direction, a multi-channel silicon photomultiplier is arranged above the microfluidic chip, a plurality of groups of detection channels for receiving different waveband fluorescence signals are arranged in the multi-channel silicon photomultiplier in an upper and lower spaced manner, a dichroic mirror, a band-pass filter and a lens group are sequentially arranged along the light path in each group of detection channels, and are collected on a receiving chip.

[0008] Further, the beam shaping module is composed of a collimating achromatic lens group and a cylindrical mirror, the collimating achromatic lens group is opposite to the excitation light source optical fiber, and the cylindrical mirror is obliquely arranged to reflect the vertical excitation light to the horizontal into the focusing and fluorescence collection module.

[0009] Further, a flow channel for receiving excitation light is arranged on the microfluidic chip.

[0010] Further, the detection channels are provided with four groups, and the dichroic mirror in the detection channel at the top is replaced by a detection mirror.

[0011] Further, the dichroic mirror and the detection mirror are both obliquely arranged to guide the fluorescence emitted by the microfluidic chip to be horizontal.

[0012] Further, the light beam shaping module is composed of a collimating achromatic lens group and a cylindrical mirror, the collimating achromatic lens group is fixed at the front end of the light beam shaping module interface, and is used for collimating the multi-wavelength light beam, and the divergence angle of the collimated excitation light beam is within 1°.

[0013] Further, the focusing and fluorescence collection module is composed of a mirror, a lenticular lens and a double cemented lens, the lenticular lens and the double cemented lens form an achromatic lens group, and the mirror is inclined in the same direction as the cylindrical mirror to guide the excitation light downward and sequentially pass through the lenticular lens and the double cemented lens and focus on the microfluidic chip.

[0014] Further, the achromatic lens group in the focusing and fluorescence collection module is used for collecting the backward fluorescence signal and transmitting the backward fluorescence signal directly from the periphery of the mirror in the focusing and fluorescence collection module to the multi-channel silicon photomultiplier detection module.

[0015] Further, the multi-channel silicon photomultiplier detection module is composed of N detection channels, each channel has a silicon photomultiplier, and different wavelengths of fluorescence signals can be independently processed. The module is also provided with N-1 dichroic mirrors and a detection mirror. In addition, each channel is also provided with a band-pass filter for filtering out light of other wavelengths and accurately extracting fluorescence signals in the required wavelength range. In addition, a lens group is arranged in each channel for adjusting the size and shape of the collected fluorescence spot to ensure that the spot can be accurately focused on the target surface of the M-SIPM, maximizing the signal-to-noise ratio of the M-SIPM.

[0016] Further, the collimating achromatic lens group of the light beam shaping module has an element aperture D in the range of 5mm to 15mm and a focal length F in the range of 10mm to 30mm.

[0017] Further, the achromatic lens group of the focusing and fluorescence collection module has an element aperture D in the range of 10mm to 25mm and a focal length F in the range of 25mm to 45mm.

[0018] Compared with the prior art, the utility model has the following beneficial effects:

[0019] Firstly, the collimating achromatic lens group and the cylindrical mirror are adopted to optimize the light path of the excitation light and the focusing efficiency.

[0020] Secondly, the excitation light source and the signal receiving system are integrated, the number of optical elements is reduced, the system structure is simplified, the optical loss is reduced, and the overall performance is improved.

[0021] Third, the system can efficiently realize multi-wavelength fluorescence detection, adapt to multi-wavelength excitation and fluorescence signal separation and reception, and is particularly suitable for complex biological analysis and cell research.

[0022] Finally, the compact design makes the module suitable for high-integration applications such as microfluidic chips, with excellent detection sensitivity and high-throughput analysis capability. The utility model not only can satisfy the demand of high sensitivity, multi-wavelength, fluorescence detection in modern biochemical analysis, also has wide application prospect, can promote the application development of microfluidic technology in the field of life science, medical diagnosis, environmental monitoring, etc. The module has great potential in the field of cell analysis, gene detection, drug screening, etc. and can provide reliable technical support for related field research and clinical application.

[0023] The utility model will be further described in detail below in combination with the drawings and specific embodiments. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 It is the structural schematic diagram of the embodiment of the utility model;

[0025] Figure 2 It is the structural composition schematic diagram of the light beam shaping module in the embodiment of the utility model;

[0026] Figure 3 It is the optical path schematic diagram of the light beam shaping module in the embodiment of the utility model;

[0027] Figure 4 It is the structural composition schematic diagram of the focusing and fluorescence collection module in the embodiment of the utility model;

[0028] Figure 5 It is the optical path schematic diagram of the focusing and fluorescence collection module in the embodiment of the utility model;

[0029] Figure 6 It is the structural composition schematic diagram of the multi-channel silicon photomultiplier in the embodiment of the utility model;

[0030] Figure 7 It is the optical path schematic diagram of the multi-channel silicon photomultiplier in the embodiment of the utility model;

[0031] Figure 8 It is the different wavelength light spot form diagram of microfluidic chip flow channel in the embodiment of the utility model.

[0032] In the figure: 101-excitation light source fiber; 102-beam shaping module; 103-focusing and fluorescence collection module; 104-microfluidic chip; 105-multichannel silicon photomultiplier; 201-collimating achromatic lens group; 202-cylindrical mirror; 401-mirror; 402-biconvex lens; 403-doublet lens; first dichroic mirror; 702-second dichroic mirror; 703-third dichroic mirror; 704-detection mirror; 705-first band-pass filter; 706-second band-pass filter; 707-third band-pass filter; 708-fourth band-pass filter; 709-first lens group; 710-second lens group; 711-third lens group; 712-fourth lens group; 713-multichannel silicon photomultiplier receiving chip. DETAILED DESCRIPTION

[0033] In order to make the above features and advantages of the utility model more obvious and easy to understand, the following specific examples are taken, and the detailed description is as follows in combination with the drawings.

[0034] As shown in the figure, a microfluidic flow cytometry fluorescence detector comprises an excitation light source fiber 101, a beam shaping module 102, a focusing and fluorescence collection module 103, a microfluidic chip 104 and a multichannel silicon photomultiplier 105. Figures 1-8

[0035] The excitation light source fiber 101 is used for inputting excitation light of different wavelengths, and its core function is to effectively guide the light source signal into the whole system. In order to ensure the transmission quality of light, a multimode optical fiber with a core diameter of 200 to 600 μm is selected, which can effectively reduce light loss and realize stable transmission of the light source signal.

[0036] In the embodiment of the utility model, the excitation light source fiber 101 is connected with the beam shaping module 102 through an adjustable flange interface, and the interface type is FC / SMA905. The design of the interface enables the user to accurately adjust the distance between the optical fiber end face and the beam shaping module 102 according to the need, so as to optimize the transmission efficiency of the light beam and the stability of the optical path.

[0037] In the embodiment of the utility model, the focusing and fluorescence collection module 103 is connected with the beam shaping module 102, which is used for focusing the excitation light in the flow channel of the microfluidic chip 104 and effectively collecting the fluorescence emission after excitation. The design of this module not only ensures that the excitation light can be accurately focused on the flow channel, but also efficiently collects and guides the generated fluorescence signal to the detection module in the next stage, that is, the multichannel silicon photomultiplier. The multichannel silicon photomultiplier 105 is used for screening the received fluorescence signal and transmitting it to different waveband signal analysis channels, wherein high-sensitivity, high-dynamic response range and low-noise silicon-based chips are arranged in different waveband channels.​

[0038] In the embodiment of the utility model, refer to the attached drawings of the specification Figure 2 The light beam shaping module provided in the embodiment comprises the collimating achromatic lens group 201 and the cylindrical mirror 202. The collimating achromatic lens group element aperture D is in the range of 5mm to 15mm, and the focal length F is in the range of 10mm to 30mm. The collimating achromatic lens group 201 is fixed to the front end of the interface.

[0039] As shown in the attached drawings of the specification Figure 3 The excitation light source fiber 101 introduces the excitation light source into the light beam shaping module 102. After collimation by the collimating achromatic lens group 201 (the divergence angle of different wavelengths is less than 1°), the light is reflected by the cylindrical mirror 202 and then transmitted to the focusing and fluorescence collection module 103.

[0040] In the embodiment of the utility model, refer to the attached drawings of the specification Figure 4 The focusing and fluorescence collection module provided in the embodiment comprises the mirror 401, the lenticular lens 402, and the double cemented lens 403. The lenticular lens 402 and the double cemented lens 403 have a lens group element aperture D in the range of 10mm to 25mm and a focal length F in the range of 25mm to 45mm, so that the light beam can be accurately focused in the flow channel of the microfluidic chip 104 at a working distance of 5mm to 20mm.

[0041] In the embodiment of the utility model, as shown in the attached drawings of the specification Figure 5 The light beam transmitted from the light beam shaping module 102 to the focusing and fluorescence collection module 103 is first reflected by the mirror 401, then incident at the center of the optical axis of the lenticular lens 402, and finally focused in the flow channel of the microfluidic chip 104 by the double cemented lens 403.

[0042] In the embodiment of the utility model, refer to the attached drawings of the specification Figure 8 The drawings show the light spot patterns at different wavelengths in the flow channel. From top to bottom, they correspond to 450nm, 490nm, and 650nm respectively. In the 450nm-650nm wavelength range, the light spot size at the microfluidic chip is 120μm-220μm in the long axis direction and 15μm-25μm in the short axis direction. Such a size design ensures that the excitation light can accurately cover the flow channel area of the microfluidic chip and uniformly excite the sample.

[0043] In the embodiment of the utility model, refer to the attached drawings of the specification Figure 6The multi-channel silicon photomultiplier provided by the embodiment comprises the first dichroic mirror 701, the second dichroic mirror 702, the third dichroic mirror 703, the detection mirror 704, the first band-pass filter 705, the second band-pass filter 706, the third band-pass filter 707, the fourth band-pass filter 708, the first lens group 709, the second lens group 710, the third lens group 711, the fourth lens group 712 and the multi-channel silicon photomultiplier receiving chip 713 arranged in sequence.

[0044] In the embodiment of the utility model, as shown in the description Figure 7 As shown in the description After the sample to be excited in the microfluidic chip 104 is excited, fluorescence is emitted, and the fluorescence is collected by the focusing and fluorescence collection module 103 first, then the fluorescence signals of different wavelengths are transmitted into the multi-channel photomultiplier 105, after the first, second and third dichroic mirrors 701, 702 and 703 and the detection mirror 704, the fluorescence signals of different wave bands are screened into different fluorescence signal detection channels, the first, second, third and fourth band-pass filters 705, 706, 707 and 708 are arranged in different channels respectively, and are used for filtering excitation light and inhibiting other wave band stray light, selectively passing through the wave band of the fluorescence signal, and the out-of-band cutoff capability is greater than or equal to 6 OD. After the filter in each channel is screened, the first, second, third and fourth lens groups are arranged in different channels, and are used for adjusting the size of the light beam incident into the multi-channel silicon photomultiplier receiving chip 713, so that the maximum receiving efficiency of the chip is met, and the signal-to-noise ratio is maximized.

[0045] The utility model is not limited to the above best implementation, and anyone can derive other various forms of microfluidic flow cytometry fluorescence detector under the enlightenment of the utility model. Any equivalent changes and modifications made in the patent application range of the utility model should belong to the coverage range of the utility model.

Claims

1. A microfluidic flow cytometric fluorescence detector characterized by: The excitation light source fiber is provided with a light beam shaping module, a focusing and fluorescence collection module and a microfluidic chip in sequence along the direction of the excitation light path of the excitation light source fiber, a multi-channel silicon photomultiplier is arranged above the microfluidic chip, a plurality of groups of detection channels for receiving different waveband fluorescence signals are arranged in the multi-channel silicon photomultiplier in an up-down interval, each group of detection channels is provided with a dichroic mirror, a band-pass filter and a lens group in sequence along the light path, and is collected on a receiving chip.

2. The microfluidic flow cytometry fluorescence detector according to claim 1, wherein: The light beam shaping module is composed of a collimating achromatic lens group and a cylindrical mirror, the collimating achromatic lens group is opposite to the excitation light source fiber, and the cylindrical mirror is obliquely arranged to reflect the vertical excitation light to the horizontal into the focusing and fluorescence collection module.

3. The microfluidic flow cytometer fluorescence detector according to claim 2, wherein: The focusing and fluorescence collection module is composed of a mirror, a lenticular lens and a double cemented lens, the mirror is inclined in the same direction as the cylindrical mirror to guide the excitation light downward and sequentially pass through the lenticular lens and the double cemented lens and focus on the microfluidic chip.

4. The microfluidic flow cytometer fluorescence detector according to claim 1, wherein: The microfluidic chip is provided with a flow channel for receiving excitation light.

5. The microfluidic flow cytometer fluorescence detector according to claim 1, wherein: The detection channel is provided with four groups, and the dichroic mirror in the top detection channel is replaced by a detection mirror.

6. The microfluidic flow cytometer fluorescence detector according to claim 5, wherein: The dichroic mirror and the detection mirror are both obliquely arranged to guide the fluorescence emitted by the microfluidic chip to be horizontal.

7. The microfluidic flow cytometer fluorescence detector of claim 1, wherein: The dichroic mirror is sequentially a first dichroic mirror, a second dichroic mirror and a third dichroic mirror from bottom to top; the band-pass filter is sequentially a first band-pass filter, a second band-pass filter, a third band-pass filter and a fourth band-pass filter from bottom to top; and the lens group is sequentially a first lens group, a second lens group, a third lens group and a fourth lens group from bottom to top.

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