Multicolor fluorescence detection device, system and method based on spatial light
By using a multi-color fluorescence detection device and method based on spatial light, and utilizing a snapshot hyperspectral camera and optical elements, the problems of spectral overlap crosstalk and filter replacement in multi-color qPCR were solved, achieving efficient and low-cost multi-color qPCR detection.
Patent Information
- Application Number
- CN202410320366.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-03-20
- Publication Date
- 2025-09-23
AI Technical Summary
Existing multi-color qPCR technology has difficulty in detecting more than seven DNA targets simultaneously, and when changing fluorescent dyes, the filter needs to be replaced, resulting in increased costs and spectral overlap and crosstalk problems.
A multi-color fluorescence detection device based on spatial light is used, with multiple reaction tubes, lighting modules, snapshot hyperspectral cameras and optical elements. By reflecting and converging fluorescence signals, the multi-spectral channels of the snapshot hyperspectral camera are used to draw the mixed fluorescence spectrum curve, and the inversion algorithm is used to demix it into monochromatic fluorescence signal intensity.
Multi-color qPCR detection with 7 or more colors is achieved with a miniaturized and low-cost device, avoiding spectral overlap and crosstalk and reducing the need for filter replacement.
Smart Images
Figure CN120682919A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the field of fluorescence detection technology, and in particular to a multi-color fluorescence detection device, system, and method based on spatial light. Background Art
[0002] Real-time fluorescence quantitative PCR (PCR) is an important method for quantitative analysis of DNA molecules, widely used in medicine and biology. The two most common methods involve adding a fluorescent dye or a fluorescent probe to a reaction tube containing the DNA to be tested. Fluorescent dyes, such as SYBR Green (a nucleic acid dye), fluoresce weakly when free but greatly enhance their fluorescence upon binding to double-stranded DNA. Fluorescent probes utilize the phenomenon of fluorescence resonance energy transfer (FRET). The probes contain a pair of groups that can undergo FRET. During enzyme cleavage and hybridization processes in the PCR reaction, the distance between the two groups changes, causing a change in fluorescence intensity. Each fluorescent dye and probe binds only to specific DNA targets, resulting in a proportional relationship between the fluorescence intensity in the tube and the amount of double-stranded DNA containing the target. Therefore, the type and amount of double-stranded DNA present in the tube can be calculated by measuring the fluorescence intensity.
[0003] The parallel amplification and detection of multiple DNA targets in a single reaction tube is called multicolor qPCR (quantitative PCR). The demand for multicolor qPCR in scientific research and clinical practice is increasing. For example, HPV testing requires quantitative analysis of 14 typing targets. Currently, multicolor qPCR technology can only detect 4-6 targets in a single tube. Because more than 7 targets require more than 7 fluorescent signals, the inevitable spectral overlap will cause serious crosstalk problems, which are difficult to eliminate using existing technology. At the same time, existing multicolor qPCR uses a separate filter for each fluorescent signal. When changing fluorescent dyes, the corresponding filter needs to be replaced at the same time, resulting in inconsistencies in usage and increased costs. Summary of the Invention
[0004] In view of this, the purpose of this application is to propose a multi-color fluorescence detection device based on spatial light that can realize multi-color qPCR detection of 7 colors or more, is miniaturized, and has low cost.
[0005] To achieve the above objectives, an embodiment of the present application discloses a multi-color fluorescence detection device based on spatial light, which includes:
[0006] a plurality of reaction tubes for containing fluorescent mixtures;
[0007] an illumination module, emitting excitation light of a corresponding wavelength to the fluorescent mixture in each reaction tube, so as to excite the fluorescent mixture to generate a corresponding fluorescent signal;
[0008] an acquisition module, comprising a second optical element, a third optical element, and a snapshot hyperspectral camera;
[0009] The second optical element reflects the fluorescent signals generated by the fluorescent mixture in the multiple reaction tubes to the third optical element, and the third optical element converges the fluorescent signals generated by the fluorescent mixture in the multiple reaction tubes to the snapshot hyperspectral camera. The snapshot hyperspectral camera converts the fluorescent signals generated by the fluorescent mixture in the multiple reaction tubes into image signals.
[0010] In one embodiment, the lighting module comprises:
[0011] A light source, a filter element and a convergence component, wherein the filter element filters the light emitted by the light source to obtain excitation light of a corresponding wavelength, and the convergence component converges the excitation light of the corresponding wavelength into each of the reaction tubes.
[0012] In one embodiment, the filter element is a bandpass filter or a beam splitter prism.
[0013] In one embodiment, the lighting module further comprises:
[0014] A first optical element is provided between the light source and the filter element. The first optical element collimates the light emitted by the light source and then emits the collimated light to the first filter element.
[0015] In one embodiment,
[0016] The converging component includes: multiple fourth optical elements and multiple fifth optical elements. The fourth optical elements, fifth optical elements and reaction tubes are arranged in a one-to-one correspondence. Each of the fourth optical elements converges the light passing through the filter element to the corresponding fifth optical element, and the fifth optical element converges the light to the corresponding reaction tube.
[0017] In one embodiment, the second optical element includes a dichroic mirror, which is arranged between the fourth optical element and the fifth optical element. The excitation light emitted from the fourth optical element passes through the first surface of the dichroic mirror and then transmits through the second surface of the dichroic mirror. The fluorescent signal generated by the fluorescent mixture in the reaction tube is reflected from the second surface of the dichroic mirror to the third optical element.
[0018] In one embodiment, the converging component includes: a sixth optical element, a plurality of illumination optical fibers, and a plurality of seventh optical elements. The illumination optical fibers, the seventh optical elements, and the reaction tubes are arranged in a one-to-one correspondence. The sixth optical element couples the light passing through the filter element into the corresponding illumination optical fiber, and each of the seventh optical elements converges the light emitted from the corresponding illumination optical fiber into the corresponding reaction tube.
[0019] In one embodiment, the second optical element includes a dichroic mirror, and the fluorescent signal generated by the fluorescent mixture in the reaction tube is reflected by the dichroic mirror to the third optical element.
[0020] In one embodiment, the light source is an LED, a mercury lamp or a laser.
[0021] In one embodiment, the snapshot hyperspectral camera includes a mosaic filter, a microlens array, and an image sensor.
[0022] Based on the same inventive concept, another embodiment of the present application discloses a multi-color fluorescence detection system based on spatial light, which includes a computer and the multi-color fluorescence detection device described in any one of the above items, wherein the computer is connected to a snapshot hyperspectral camera in the multi-color fluorescence detection device, and the computer receives and records the image signal output by the snapshot hyperspectral camera.
[0023] Based on the same inventive concept, another embodiment of the present application discloses a multi-color fluorescence detection method based on spatial light, which is applied to any of the multi-color fluorescence detection devices described above. The method includes:
[0024] Drawing a multi-color mixed fluorescence spectrum curve according to the image signal output by the snapshot hyperspectral camera;
[0025] The mixed fluorescence spectrum curve is unmixed into a single-color fluorescence spectrum curve using an inversion algorithm to obtain the relative intensity of each single-color fluorescence signal.
[0026] In one embodiment, the step of drawing a multi-color mixed fluorescence spectrum curve based on the image signal output by the snapshot hyperspectral camera includes:
[0027] Obtaining the image signal intensity output by each spectral channel of the snapshot hyperspectral camera;
[0028] The image signal intensity output by the first spectral channel and the image signal intensity output by the last spectral channel are connected, and the multi-color mixed fluorescence spectrum curve is obtained by fitting.
[0029] In one embodiment, unmixing the mixed fluorescence spectrum curve into a monochromatic fluorescence spectrum curve using an inversion algorithm to obtain the relative intensity of each monochromatic fluorescence signal includes:
[0030] Construct the following formula:
[0031]
[0032] Wherein, N is the total number of spectral channels in the snapshot hyperspectral camera, D1, D2...D N is the intensity of each monochromatic fluorescence signal obtained for each spectral channel; J is the number of fluorescent dyes; the fluorescence signal intensity emitted by each fluorescent dye is I1, I2...I J Wherein, when the fluorescence emitted by the first fluorescent dye is imaged by the snapshot hyperspectral camera alone, the normalized intensities of the N spectral channels are F 11 , F 21 ,……,F N1 ; F 11 and F N1 The monochromatic fluorescence spectrum curve of the first fluorescent dye is obtained by connecting and fitting. When the fluorescence emitted by the Jth dye is imaged by the snapshot hyperspectral camera alone, the normalized intensities of the N spectral channels are F 1J , F 2J ,……,F NJ ;
[0033] Solve I1 to I J , and obtain the relative intensity of each monochromatic fluorescence signal.
[0034] The spatial light-based multicolor fluorescence detection device disclosed in this application uses a second optical element to reflect the fluorescence signal generated by the fluorescent mixture in a reaction tube to a third optical element. The third optical element then converges the fluorescence signals generated by the fluorescent mixtures in multiple reaction tubes and couples them into a snapshot hyperspectral camera. Using the 20 to thousands of spectral channels within the snapshot hyperspectral camera, a spectral curve of the detected multicolor mixed fluorescence signal can be plotted, thereby determining the relative intensities of each monochromatic fluorescence signal. This device can achieve multicolor qPCR detection with seven or more colors, offering the advantages of miniaturization and low cost. BRIEF DESCRIPTION OF THE DRAWINGS
[0035] Figure 1 A schematic structural diagram of a multi-color fluorescence detection device based on spatial light provided in one embodiment of the present application;
[0036] Figure 2 A schematic structural diagram of a multi-color fluorescence detection device based on spatial light provided in another embodiment of the present application;
[0037] Figure 3A schematic structural diagram of a snapshot hyperspectral camera provided in one embodiment of the present application;
[0038] Figure 4 A schematic structural diagram of a snapshot hyperspectral camera is provided for another embodiment of the present application;
[0039] Figure 5 A schematic structural diagram of a mosaic filter provided in one embodiment of the present application;
[0040] Figure 6 A flowchart of a multi-color fluorescence detection method based on spatial light provided in one embodiment of the present application;
[0041] Figure 7 This is a flowchart of a multi-color fluorescence detection method based on spatial light provided in another embodiment of the present application.
[0042] Marking Description:
[0043] 10. Reaction tube; 21. Light source; 22. Filter element; 23. First optical element; 24. Fourth optical element; 25. Fifth optical element; 26. Sixth optical element; 27. Illumination fiber; 28. Seventh optical element; 31. Second optical element; 32. Third optical element; 33. Snapshot hyperspectral camera; 341. Mosaic filter; 342. Microlens array; 343. Image sensor; 344. First convex lens; 345. Second convex lens. DETAILED DESCRIPTION
[0044] In order to make the objectives, technical solutions and advantages of this application more clear, this application is further described in detail below in combination with specific implementation methods and with reference to the accompanying drawings.
[0045] It should be noted that, unless otherwise defined, the technical terms or scientific terms used in the embodiments of the present application should have the usual meanings understood by people with ordinary skills in the field to which the present application belongs. The "first", "second" and similar words used in the embodiments of the present application do not indicate any order, quantity or importance, but are only used to distinguish different components. "Include" or "comprise" and similar words mean that the elements or objects appearing before the word include the elements or objects listed after the word and their equivalents, without excluding other elements or objects. "Connect" or "connected" and similar words are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.
[0046] like Figure 1 and 2As shown, the embodiment of the present application provides a multi-color fluorescence detection device based on spatial light, which includes:
[0047] A plurality of reaction tubes 10 for containing a fluorescent mixture;
[0048] An illumination module emits excitation light of a corresponding wavelength to the fluorescent mixture in each reaction tube 10 to excite the fluorescent mixture to generate a corresponding fluorescent signal;
[0049] An acquisition module, comprising a second optical element 31, a third optical element 32 and a snapshot hyperspectral camera 33;
[0050] Among them, the second optical element 31 reflects the fluorescent signal generated by the fluorescent mixture in the multiple reaction tubes to the third optical element 32, the third optical element 32 converges the fluorescent signal generated by the fluorescent mixture in the multiple reaction tubes 10 and couples it into the snapshot hyperspectral camera 33, and the snapshot hyperspectral camera 33 converts the fluorescent signal generated by the fluorescent mixture in the multiple reaction tubes 10 into an image signal.
[0051] The number of reaction tubes 10 can be 20, 48, 96, etc., without any specific limitation.
[0052] The spatial light-based multicolor fluorescence detection device provided in this embodiment utilizes a second optical element 31 to reflect the fluorescence signal generated by the fluorescent mixture within a reaction tube 10 to a third optical element 32. The third optical element 32 then converges the fluorescence signals generated by the fluorescent mixtures within multiple reaction tubes 10 and couples them into a snapshot hyperspectral camera 33. Using the 20 to thousands of spectral channels within the snapshot hyperspectral camera 33, a spectral curve of the detected multicolor mixed fluorescence signal can be plotted, thereby determining the relative intensities of each monochromatic fluorescence signal. This device can achieve multicolor qPCR detection with seven or more colors, offering the advantages of miniaturization and low cost.
[0053] Furthermore, the multi-color fluorescence detection device based on spatial light provided in this embodiment also includes a thermal cycle heating and cooling module, which is composed of a temperature controller, a temperature sensor, a heating and cooling module, a radiator, etc.
[0054] In one embodiment, the illumination module includes a light source 21, a filter element 22, and a focusing assembly. The filter element 22 filters the light emitted by the light source 21 to obtain excitation light of a corresponding wavelength. The focusing assembly focuses the excitation light of the corresponding wavelength into each reaction tube 10. The filter element 22 is used to filter out light other than the excitation light. Optionally, the filter element 22 is a bandpass filter or a beam splitter prism; the light source 21 is an LED, a mercury lamp, or a laser.
[0055] Reference Figure 1As shown, in one embodiment, the illumination module further includes a first optical element 23, which is disposed between the light source 21 and the filter element 22. The first optical element 23 collimates the light emitted by the light source 21 and transmits it to the filter element 22. The first optical element 23 ensures that the excitation light energy irradiated to different reaction tubes 10 is equal, that is, the excitation conditions of different reaction tubes 10 are the same. Optionally, the first optical element 23 is a convex lens or a Fresnel lens.
[0056] Please continue to refer to Figure 1 As shown, in one embodiment, the converging assembly includes: a plurality of fourth optical elements 24 and a plurality of fifth optical elements 25. The fourth optical elements 24, the fifth optical elements 25, and the reaction tubes 10 are arranged in a one-to-one correspondence. Each fourth optical element 24 converges the light passing through the filter element 22 to the corresponding fifth optical element 25, and the fifth optical element 25 converges the light to the corresponding reaction tube 10. Optionally, the fourth optical element 24 is a convex lens or a Fresnel lens, and the fifth optical element 25 is a convex lens or a Fresnel lens.
[0057] Furthermore, the second optical element 31 includes a dichroic mirror, which is disposed between the fourth optical element 24 and the fifth optical element 25. The excitation light emitted from the fourth optical element 24 passes through the first surface of the dichroic mirror and then transmits through the second surface of the dichroic mirror. The fluorescent signal generated by the fluorescent mixture in the reaction tube 10 is reflected by the second surface of the dichroic mirror to the third optical element 32. In other words, the illumination light path and the collection light path share the dichroic mirror, which helps reduce the use of optical components.
[0058] Reference Figure 2 As shown, in another embodiment, the converging assembly includes a sixth optical element 26, a plurality of illumination fibers 27, and a plurality of seventh optical elements 28. The illumination fibers 27, the seventh optical elements 28, and the reaction tubes 10 are disposed in a one-to-one correspondence. The sixth optical element 26 couples the light passing through the optical filter element 22 into the corresponding illumination fiber, and each seventh optical element 28 converges the light emitted from the corresponding illumination fiber 27 into the corresponding reaction tube 10. Optionally, the sixth optical element 26 is a convex lens or a Fresnel lens, and the seventh optical element 28 is a convex lens or a Fresnel lens.
[0059] Furthermore, the second optical element 31 includes a dichroic mirror, and the fluorescent signal generated by the fluorescent mixture in the reaction tube 10 is reflected to the third optical element 32 through the dichroic mirror.
[0060] Reference Figure 3-5 As shown, the snapshot hyperspectral camera 33 includes a mosaic filter 341, a microlens array 342 and an image sensor 343. Figure 3As shown in one embodiment, the snapshot hyperspectral camera 33 includes two microlens arrays 342. The fluorescence signal passes through the mosaic filter 341 in sequence and reaches the first microlens array 342 and the second microlens array 342 and converges to the image sensor 343. Figure 4 As shown, in another embodiment, the snapshot hyperspectral camera 33 includes a telescope optical system, which includes a first convex lens 344 and a second convex lens 345. The fluorescence signal is collimated by the first convex lens 344 and the second convex lens 345 in sequence, and then passes through the mosaic filter 341 before being converged by the microlens array 342 to the image sensor 343.
[0061] The snapshot hyperspectral camera 33 provides 20 to thousands of spectral channels, plotting the spectral curve of the detected multicolor mixed fluorescence signal. Based on the known spectral curves of each single-color fluorescence signal, an inversion algorithm accurately restores the relative intensities of each single-color fluorescence signal. This solves the problem in existing multicolor qPCR detection equipment where each filter can only sample the peak intensity region of a single-color fluorescence signal, and where overlapping peak regions of two fluorescence signals can cause severe crosstalk, enabling multicolor qPCR detection of seven or more colors.
[0062] The snapshot hyperspectral camera 33 can simultaneously image the spectral region of interest, such as the visible light band where most of the fluorescence signals are located, so when the fluorescent dye used is changed, there is no need to change the filter.
[0063] Among them, the mosaic filter 341 has multiple implementation methods (absorption filter, interference filter, induced transmission filter (ITF), linear gradient filter, surface plasmon filter, metasurface structure filter). Their common feature is that they transmit spectral information of different bands at different positions in space, and then use array-type photoelectric sensors to analyze the signals at different positions in space, thereby obtaining the spectral information of the object in each spectral band.
[0064] In one embodiment, the snapshot hyperspectral camera 33 includes a 60-channel mosaic filter, which operates on the principle of FP cavity interferometry. Due to its low optical density (OD), some excitation light will pass through and cause crosstalk with the fluorescence. Therefore, an additional bandpass filter is required to filter out the excitation light and allow only the fluorescence signal to pass.
[0065] Another embodiment of the present application provides a multi-color fluorescence detection system based on spatial light, which includes a computer and the multi-color fluorescence detection device in the above embodiment. The computer is connected to a snapshot hyperspectral camera in the multi-color fluorescence detection device, and the computer receives and records the image signal output by the snapshot hyperspectral camera.
[0066] Reference Figure 6As shown, another embodiment of the present application provides a multi-color fluorescence detection method based on spatial light, which is applied to the multi-color fluorescence detection device in the above embodiment. The method includes the following steps:
[0067] Step S10, drawing a multi-color mixed fluorescence spectrum curve based on the image signal output by the snapshot hyperspectral camera;
[0068] Step S20: using an inversion algorithm to unmix the mixed fluorescence spectrum curve into a single-color fluorescence spectrum curve, and obtaining the relative intensity of each single-color fluorescence signal.
[0069] Reference Figure 7 As shown, in one embodiment, step S10, drawing a multi-color mixed fluorescence spectrum curve based on the image signal output by the snapshot hyperspectral camera 34, includes:
[0070] Step S11, obtaining the image signal intensity output by each spectral channel of the snapshot hyperspectral camera;
[0071] Step S12: Connect the image signal intensity output by the first spectral channel and the image signal intensity output by the last spectral channel, and fit them to obtain a multi-color mixed fluorescence spectrum curve.
[0072] In one embodiment, step S20, using an inversion algorithm to unmix the mixed fluorescence spectrum curve into a single-color fluorescence spectrum curve to obtain the relative intensity of each single-color fluorescence signal, includes:
[0073] Construct the following formula:
[0074]
[0075] Where N is the total number of spectral channels in the snapshot hyperspectral camera 33, D1, D2...D N is the intensity of each monochromatic fluorescence signal obtained for each spectral channel; J is the number of fluorescent dyes; the fluorescence signal intensity emitted by each fluorescent dye is I1, I2...I J When the fluorescence emitted by the first fluorescent dye is imaged by the snapshot hyperspectral camera 33 alone, the normalized intensities of the N spectral channels are F 11 , F 21 ,……,F N1 ; F 11 and F N1 The monochromatic fluorescence spectrum curve of the first fluorescent dye is obtained by connecting and fitting. When the fluorescence emitted by the Jth dye is imaged by the snapshot hyperspectral camera alone, the normalized intensities of the N spectral channels are F 1J , F 2J ,……,F NJ ;
[0076] Solve I1 to I J , and obtain the relative intensity of each monochromatic fluorescence signal.
[0077] This multicolor fluorescence detection method based on spatial light solves the problem in existing multicolor qPCR detection equipment where each filter can only sample the peak intensity region of a single-color fluorescence signal, and severe crosstalk occurs when two fluorescence signal peak regions overlap. This method enables multicolor qPCR detection with seven or more colors.
[0078] It should be noted that "a certain body" or "a certain part" can be a part of the corresponding "component", that is, the "a certain body" or "a certain part" can be integrally formed with the "other parts of the component"; or it can be an independent component that is separable from the "other parts of the component", that is, the "a certain body" or "a certain part" can be manufactured independently and then combined with the "other parts of the component" into a whole. The expression of the above-mentioned "a certain body" or "a certain part" in this application is only one of the implementation methods, for the convenience of reading, and not to limit the scope of protection of this application. As long as it contains the above-mentioned features and has the same function, it should be understood as an equivalent technical solution of this application.
[0079] It should be noted that the above description only describes some embodiments of the present application. Other embodiments are within the scope of the appended claims. In some cases, the actions or steps recited in the claims can be performed in a different order than described in the above embodiments and still achieve the desired results. Furthermore, the processes depicted in the accompanying drawings do not necessarily require the specific order or sequential order shown to achieve the desired results.
[0080] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present application (including the claims) is limited to these examples. Within the scope of the present application, the above embodiments or technical features in different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of different aspects of the embodiments of the present application as described above, which are not provided in detail for the sake of simplicity.
[0081] The embodiments of the present application are intended to encompass all such substitutions, modifications, and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the embodiments of the present application should be included in the scope of protection of this application.
Claims
1. A multi-color fluorescence detection device based on spatial light, characterized in that: include: a plurality of reaction tubes for containing fluorescent mixtures; an illumination module, emitting excitation light of a corresponding wavelength to the fluorescent mixture in each reaction tube, so as to excite the fluorescent mixture to generate a corresponding fluorescent signal; an acquisition module, comprising a second optical element, a third optical element, and a snapshot hyperspectral camera; The second optical element reflects the fluorescent signals generated by the fluorescent mixture in the multiple reaction tubes to the third optical element, and the third optical element converges the fluorescent signals generated by the fluorescent mixture in the multiple reaction tubes to the snapshot hyperspectral camera. The snapshot hyperspectral camera converts the fluorescent signals generated by the fluorescent mixture in the multiple reaction tubes into image signals.
2. The multi-color fluorescence detection device based on spatial light according to claim 1, characterized in that: The lighting module comprises: A light source, a filter element and a convergence component, wherein the filter element filters the light emitted by the light source to obtain excitation light of a corresponding wavelength, and the convergence component converges the excitation light of the corresponding wavelength into each of the reaction tubes.
3. The multi-color fluorescence detection device based on spatial light according to claim 2, characterized in that: The filter element is a bandpass filter or a spectroscopic prism.
4. The multi-color fluorescence detection device based on spatial light according to claim 2, characterized in that: The lighting module further comprises: A first optical element is provided between the light source and the filter element, and the first optical element collimates the light emitted by the light source and then emits the collimated light to the filter element.
5. The multi-color fluorescence detection device based on spatial light according to claim 2, characterized in that: The convergence component includes: A plurality of fourth optical elements and a plurality of fifth optical elements are provided, wherein the fourth optical elements, the fifth optical elements and the reaction tubes are arranged in a one-to-one correspondence, each of the fourth optical elements converges the light passing through the filter element to the corresponding fifth optical element, and the fifth optical element converges the light to the corresponding reaction tube.
6. The multi-color fluorescence detection device based on spatial light according to claim 5, characterized in that: The second optical element includes a dichroic mirror, which is arranged between the fourth optical element and the fifth optical element. The excitation light emitted from the fourth optical element passes through the first surface of the dichroic mirror and then transmits through the second surface of the dichroic mirror. The fluorescent signal generated by the fluorescent mixture in the reaction tube is reflected from the second surface of the dichroic mirror to the third optical element.
7. The multi-color fluorescence detection device based on spatial light according to claim 2, characterized in that: The convergence component includes: A sixth optical element, a plurality of illumination optical fibers, and a plurality of seventh optical elements, wherein the illumination optical fibers, the seventh optical elements, and the reaction tubes are arranged in a one-to-one correspondence, the sixth optical element couples the light passing through the filter element into the corresponding illumination optical fiber, and each of the seventh optical elements converges the light emitted from the corresponding illumination optical fiber into the corresponding reaction tube.
8. The multi-color fluorescence detection device based on spatial light according to claim 7, characterized in that: The second optical element includes a dichroic mirror, and the fluorescent signal generated by the fluorescent mixture in the reaction tube is reflected by the dichroic mirror to the third optical element.
9. The multi-color fluorescence detection device based on spatial light according to claim 2, characterized in that: The light source is an LED, a mercury lamp or a laser.
10. The multi-color fluorescence detection device based on spatial light according to claim 1, characterized in that: The snapshot hyperspectral camera includes a mosaic filter, a microlens array and an image sensor.
11. A multi-color fluorescence detection system based on spatial light, characterized in that: The method comprises a computer and the multi-color fluorescence detection device according to any one of claims 1 to 10, wherein the computer is connected to a snapshot hyperspectral camera in the multi-color fluorescence detection device, and the computer receives and records image signals output by the snapshot hyperspectral camera.
12. A multi-color fluorescence detection method based on spatial light, applied to the multi-color fluorescence detection device according to any one of claims 1 to 10, characterized in that: The method comprises: Drawing a multi-color mixed fluorescence spectrum curve according to the image signal output by the snapshot hyperspectral camera; The mixed fluorescence spectrum curve is unmixed into a single-color fluorescence spectrum curve using an inversion algorithm to obtain the relative intensity of each single-color fluorescence signal.
13. The multi-color fluorescence detection method based on spatial light according to claim 12, characterized in that: Drawing a multi-color mixed fluorescence spectrum curve based on the image signal output by the snapshot hyperspectral camera includes: Obtaining the image signal intensity output by each spectral channel of the snapshot hyperspectral camera; The image signal intensity output by the first spectral channel and the image signal intensity output by the last spectral channel are connected, and the multi-color mixed fluorescence spectrum curve is obtained by fitting.
14. The multi-color fluorescence detection method based on spatial light according to claim 12, characterized in that: The method of unmixing the mixed fluorescence spectrum curve into a monochromatic fluorescence spectrum curve by using an inversion algorithm to obtain the relative intensity of each monochromatic fluorescence signal includes: Construct the following formula: Wherein, N is the total number of spectral channels in the snapshot hyperspectral camera, D1, D2...D N is the intensity of each monochromatic fluorescence signal obtained for each spectral channel; J is the number of fluorescent dyes; the fluorescence signal intensity emitted by each fluorescent dye is I1, I2...I J Wherein, when the fluorescence emitted by the first fluorescent dye is imaged by the snapshot hyperspectral camera alone, the normalized intensities of the N spectral channels are F 11 , F 21 ,……,F N1 ; F 11 and F N1 The monochromatic fluorescence spectrum curve of the first fluorescent dye is obtained by connecting and fitting. When the fluorescence emitted by the Jth dye is imaged by the snapshot hyperspectral camera alone, the normalized intensities of the N spectral channels are F 1J , F 2J ,……,F NJ ; Solve I1 to I J , and obtain the relative intensity of each monochromatic fluorescence signal.