Fluorescent quantitative detection device and fluorescent quantitative detection system
By using a fluorescence quantitative detection device that combines excitation fiber and fluorescence collection fiber, simultaneous detection of multiple samples is achieved, solving the problems of low quantitative accuracy and efficiency, and improving detection precision and speed.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- SHANGHAI SAILU LIFE SCIENCES CO LTD
- Filing Date
- 2025-07-18
- Publication Date
- 2026-06-26
Smart Images
Figure CN224411768U_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of biological detection technology, and in particular to a fluorescence quantitative detection device and a fluorescence quantitative detection system. Background Technology
[0002] Before second-generation sequencing, DNA needs to be extracted and libraries constructed. At the end of the library construction process, the sample concentration needs to be quantified. Then, samples of different concentrations are diluted to the same concentration according to the quantification results, and pooling (a technique for mixing multiple samples or libraries for unified sequencing) is performed before sequencing. Therefore, the accuracy of quantification has a crucial impact on the final sequencing data volume of different samples.
[0003] In the process of developing this application, the inventors discovered that the prior art has at least the following problems:
[0004] In traditional quantitative methods, excitation light passes through the wall of the quantitative quantification tube to excite the sample inside. The sample fluorescence then passes through the tube wall again before being received by the detector. Since both excitation and fluorescence emission require passage through the quantitative quantification tube, its optical parameters, such as transmittance and scattering rate, affect the accuracy of fluorescence quantification. Because quantitative quantification tubes are consumables, the optical parameters of different tubes cannot be strictly kept consistent, leading to fluctuations in the quantitative results and impacting accuracy.
[0005] Secondly, in terms of quantitative throughput, traditional methods mostly rely on single-strip quantification. When dealing with large sample sizes, such as 96 samples, the quantification cycle is lengthy and inefficient. While current technologies have introduced eight-strip quantification schemes, improving efficiency, they still require twelve manual operations for 96 samples, making the process cumbersome and prone to errors. Summary of the Invention
[0006] Based on this, this application provides a fluorescence quantitative detection device and a fluorescence quantitative detection system to improve the problems of low quantitative accuracy and low quantitative detection efficiency in the prior art.
[0007] To achieve the above objectives, the technical solution of this application embodiment is implemented as follows:
[0008] On one hand, embodiments of this application provide a fluorescence quantitative detection device, including a laser, an excitation fiber, an excitation microlens array, a fluorescence collection fiber, a signal collection microlens array, and an imaging system;
[0009] The excitation fiber includes multiple excitation light output ends. The excitation fiber is used to receive the excitation light emitted by the laser and output the excitation light from the excitation light output ends to the excitation microlens array.
[0010] The excitation microlens array includes multiple excitation microlens units, and the excitation light microlens array is used to focus the excitation light onto the solutions in each sample quantitative tube containing the solution to be tested;
[0011] The fluorescence collecting optical fiber includes multiple fluorescence input ends and multiple fluorescence output ends. One fluorescence input end and a corresponding excitation output end are arranged side by side and are arranged side by side at the front focal point of a corresponding excitation microlens unit. The fluorescence collecting optical fiber is used to collect the fluorescence signal of the test solution and output the fluorescence signal from the fluorescence output end to the collecting microlens array.
[0012] The signal-collecting microlens array is used to image different regions of the imaging system.
[0013] In one embodiment, the excitation fiber includes an excitation light input segment and an excitation light output segment arranged sequentially. The excitation light input segment is located near the excitation light input end and is a fiber bundle structure; the excitation light output segment is located near the excitation light output end and is a single fiber structure.
[0014] In one embodiment, the fluorescence collecting optical fiber is provided with a fluorescence collecting section, a fluorescence transmission section and a fluorescence output section in sequence. The fluorescence transmission section is an optical fiber bundle structure. The fluorescence collecting section is located near the fluorescence input end, and the fluorescence output section is located near the fluorescence output end. The fluorescence collecting section and the fluorescence output section are both single optical fiber structures.
[0015] In one embodiment, the excitation microlens array includes a plurality of excitation microlens units arranged in an array, and the signal collection microlens array includes a plurality of signal collection microlens units arranged in an array.
[0016] The number of excitation light output terminals, the number of fluorescence input terminals, the number of fluorescence output terminals, the number of excitation microlens units, and the number of signal collection microlens units are all the same as the number of sample quantitative tubes.
[0017] In one embodiment, the front focal point of the excitation microlens unit is located above the sample quantification tube, so that the excitation light enters the test solution directly from above the opening of the sample quantification tube via the excitation microlens unit.
[0018] In one embodiment, the back focus of the excitation microlens unit is located inside the test solution within the sample quantification tube.
[0019] In one embodiment, the imaging system includes an imaging device and a fluorescence filter, the fluorescence filter being disposed between the fluorescence output end and the imaging device, the imaging device being used to receive the image array output by the signal collection microlens array, and to output the image array at once.
[0020] In one embodiment, the imaging device includes an area array camera or a PMT array photodetector.
[0021] In one embodiment, the focal length of the excitation microlens unit is in the range of 20-30 mm.
[0022] On the other hand, embodiments of this application provide a fluorescence quantitative detection system, including the fluorescence quantitative detection device as described above.
[0023] This application has at least the following beneficial effects: The fluorescence quantitative detection device and system provided in this application include multiple excitation light output ends, multiple fluorescence input ends, and multiple fluorescence output ends, enabling simultaneous detection of multiple samples and greatly improving the efficiency of quantitative detection. The fluorescence quantitative detection device provided in this application transmits excitation light and fluorescence signals through excitation optical fibers and fluorescence collection optical fibers, unaffected by the optical parameters of the quantitative tube, effectively improving the accuracy of quantitative detection. Placing the fluorescence input ends and excitation light output ends side-by-side at the front focal point of the excitation microlens unit effectively improves the input efficiency of the excitation light and the collection efficiency of the fluorescence signal. The fluorescence signal is ultimately output to different areas of the imaging system through the collection microlens array, achieving synchronous imaging of all fluorescence signals and greatly improving the efficiency of quantitative detection. Attached Figure Description
[0024] Figure 1 This is a schematic diagram of the structure of the fluorescence quantitative detection device according to an embodiment of this application.
[0025] The meanings of the labels in the attached diagram are as follows:
[0026] 1. Excitation light emission system; 11. Laser; 12. Excitation coupling lens;
[0027] 2. Excitation fiber; 21. Excitation light input section; 22. Excitation light output section;
[0028] 3. Excite the microlens unit;
[0029] 4. Fluorescence collection fiber; 41. Fluorescence collection section; 42. Fluorescence transmission section; 43. Fluorescence output section;
[0030] 5. Signal collection microlens unit;
[0031] 6. Imaging system; 61. Imaging equipment; 62. Fluorescent filter;
[0032] 7. Sample quantification tube. Detailed Implementation
[0033] The technical solution of this application will be further described in detail below with reference to the accompanying drawings and specific embodiments.
[0034] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for the purpose of describing particular embodiments only and is not intended to limit the ways in which this application may be implemented. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.
[0035] In the description of this application, it should be understood that the terms "center," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," and "outer," etc., indicating the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, are used only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on this application. In the description of this application, unless otherwise stated, "a plurality of" means two or more.
[0036] In the description of this application, it should be noted that, unless otherwise expressly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal communication between two components. Those skilled in the art can understand the specific meaning of the above terms in this application based on the specific circumstances.
[0037] Please see Figure 1 The fluorescence quantitative detection device of this application includes a laser, an excitation fiber 2, an excitation microlens array, a fluorescence collection fiber 4, a signal collection microlens array, and an imaging system 6.
[0038] The excitation fiber 2 includes multiple excitation light output ends. The excitation fiber 2 is used to receive excitation light and output the excitation light emitted by the laser from the excitation light output ends to the excitation microlens array.
[0039] The excitation microlens array comprises multiple excitation microlens units 3 arranged in an array. The excitation microlens array is used to focus the excitation light onto the solutions in each sample quantitative tube 7 containing the solution to be tested. The focal range of the excitation microlens unit 3 is 20-30 mm.
[0040] The fluorescence collecting fiber 4 includes multiple fluorescence input ends and multiple fluorescence output ends. One fluorescence input end and one corresponding excitation output end are arranged in parallel and are arranged in parallel at the front focal point of the corresponding excitation microlens unit 3. The fluorescence collecting fiber 4 is used to collect the fluorescence signal of the solution to be tested and output the fluorescence signal from the fluorescence output end to the collecting microlens array.
[0041] The signal collecting microlens array includes multiple signal collecting microlens units 5 arranged in an array, which are used to image different regions of the imaging system 6 respectively.
[0042] The fluorescence quantitative detection device of this embodiment includes an excitation light emission system 1, which outputs excitation light to the excitation light output end. Specifically, the excitation light emission system 1 may include, for example, the laser 11 and the excitation coupling lens 12 described above. The laser 11 emits a parallel beam, which is focused by the excitation coupling lens 12 to the input end of the excitation fiber 2. The wavelengths of the excitation light emitted by the laser 11 include 460 nm and 635 nm, and the two wavelengths of excitation light are used to excite DNA and RNA, respectively.
[0043] The excitation fiber 2 in this embodiment includes an excitation light input section 21 and an excitation light output section 22 arranged sequentially. The excitation light input section 21 is located near the excitation light input end and is a fiber bundle structure. The excitation light output section 22 is located near the excitation light output end and is a single fiber structure. That is, the excitation fiber 2 in this embodiment is roughly divided into two sections from the middle. The section near the laser 11 is the excitation light input section 21, in which multiple single fibers are bundled together; the section near the excitation microlens array is the excitation light output section 22, in which multiple single fibers are scattered single fiber structures. The end face of the excitation light input end is located at the rear focal point of the excitation coupling lens 12, and the end face of each excitation light output end is located at the front focal point of the corresponding excitation microlens unit 3.
[0044] In this embodiment, the test solutions are placed in sample quantitative tubes 7, and the test solutions in each sample quantitative tube 7 are located at the back focal point of a corresponding excitation microlens unit 3.
[0045] In this embodiment, the fluorescence collecting fiber 4 is sequentially provided with a fluorescence collecting section 41, a fluorescence transmission section 42, and a fluorescence output section 43. The fluorescence transmission section 42 is a fiber bundle structure. The fluorescence collecting section 41 is located near the fluorescence input end, and the fluorescence output section 43 is located near the fluorescence output end. Both the fluorescence collecting section 41 and the fluorescence output section 43 are single-fiber structures. That is, the fluorescence collecting fiber 4 is divided into three sections. The fluorescence transmission section 42 is located in the middle and is a fiber bundle formed by multiple single fibers. The section near the excitation microlens array is the fluorescence collecting section 41, and the section near the signal collecting microlens array is the fluorescence output section 43. Both of these sections are single-fiber structures with multiple single fibers in a scattered state. The end face of one fluorescence input end and the end face of one excitation light input end are arranged side by side at the front focal point of a corresponding excitation microlens unit 3. The end face of each fluorescence output end is located at the front focal point of a corresponding signal collecting microlens unit 5.
[0046] The front focal point of the excitation microlens unit 3 is located above the sample quantitative tube 7, allowing the excitation light to directly enter the test solution from above the opening of the sample quantitative tube 7 via the excitation microlens unit 3. The rear focal point of the excitation microlens unit 3 is located inside the test solution within the sample quantitative tube 7. The excitation light output end of the excitation fiber 2 and the fluorescence input end of the fluorescence collection fiber 4 are respectively located above the sample quantitative tube 7, ensuring that the transmission of excitation light and fluorescence signals does not need to pass through the tube wall of the sample quantitative tube 7. Therefore, the optical parameters of the sample quantitative tube 7 will not affect the final detection result, which is beneficial to improving the accuracy of quantitative detection. Simultaneously, placing the fluorescence input end and the excitation light output end side-by-side at the front focal point of the excitation microlens unit 3 can effectively improve the input efficiency of the excitation light and the collection efficiency of the fluorescence signal. The imaging system 6 of this embodiment includes an imaging device 61 and a fluorescence filter 62. The fluorescence filter 62 is disposed between the fluorescence output end and the imaging device 61. The imaging device 61 is used to receive the image array output by the signal collection microlens array and to output the image array in a single operation. The imaging device 61 includes an area array camera or a PMT array photodetector. The receiving end of the imaging device 61 is located on the plane formed by the back focal point of the multiple signal collecting microlens units 5 of the collecting microlens array.
[0047] In this embodiment, the number of excitation light output terminals, fluorescence input terminals, fluorescence output terminals, excitation microlens units 3, and signal collection microlens units 5 are all the same as the number of sample quantitative tubes 7. For example, in some embodiments, the number of samples of the test solution is 96 (i.e., the number of sample quantitative tubes 7 is 96), then the corresponding excitation fiber 2 includes 96 single optical fibers, the fluorescence collection fiber 4 also includes 96 single optical fibers, and the number of excitation microlens units 3 and signal collection microlens units 5 are also 96 each, all consistent with the number of sample quantitative tubes 7. This facilitates the quantitative detection of all test solution samples at once, thereby improving the efficiency of quantitative detection.
[0048] The working process of the fluorescence quantitative detection device in this embodiment is as follows: The laser 11 emits parallel excitation light, which is converged to the input end of the excitation fiber by the excitation coupling lens 12. The excitation fiber bundle consists of 96 single fibers. Then, the excitation light reaches the excitation microlens array above the 96 sample quantitative tubes 7 through the 96 single fibers, and is focused into the corresponding test solution after passing through the 96 excitation microlens units 3.
[0049] The fluorescence signals are transmitted to the signal detection end (fluorescence output end) through the fluorescence input ends of 96 fluorescence collecting optical fibers 4. Each of the 96 single optical fibers at the fluorescence output end corresponds to a signal collecting microlens unit 5. After passing through the 96 signal collecting microlens units 5, the fluorescence signals are converged to different positions of the imaging device 61 for one-time imaging.
[0050] This application also provides a fluorescence quantitative detection system, including the fluorescence quantitative detection device described in the above embodiments.
[0051] To avoid the influence of the optical parameters of the sample quantitative tube material on the quantitative results, the fluorescence quantitative detection device in this application adopts an epi-emission excitation and signal acquisition scheme, which avoids the influence of the sample quantitative tube in the optical path. Furthermore, for system compactness, both the excitation and collection optical paths are made of optical fibers.
[0052] In terms of high-throughput real-time detection, this application employs a combination of excitation optical fibers and fluorescence collection optical fibers. When detecting 96 samples of the solution to be tested, the total number of optical fibers in the excitation fiber is 96, with the excitation light output ends placed at the front focal point of the excitation microlens array. The output light from each fiber passes through its corresponding excitation microlens unit and is then focused into the sample quantification tube below.
[0053] Near the focal point of the excitation microlens array, a fluorescence collecting fiber is also placed, with one fiber corresponding to each sample quantification tube. These fibers are bundled in the middle section and split into 96 individual fibers at the front and rear sections. The fluorescence output from each fiber passes through the corresponding microlens, then through a fluorescence filter, and finally converges onto the imaging device. Therefore, this scheme avoids the influence of the sample quantification tube on the quantification accuracy while enabling the quantification of 96 samples at once, achieving high quantification accuracy and fast quantification speed, effectively improving quantification efficiency.
[0054] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.
[0055] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.
Claims
1. A fluorescence quantitative detection device, characterized in that, It includes a laser, an excitation fiber, an excitation microlens array, a fluorescence collection fiber, a signal collection microlens array, and an imaging system; The excitation fiber includes multiple excitation light output ends. The excitation fiber is used to receive the excitation light emitted by the laser and output the excitation light from the excitation light output ends to the excitation microlens array. The excitation microlens array includes multiple excitation microlens units, and the excitation light microlens array is used to focus the excitation light onto the solutions in each sample quantitative tube containing the solution to be tested; The fluorescence collecting fiber includes multiple fluorescence input ends and multiple fluorescence output ends. One fluorescence input end and a corresponding excitation light output end are arranged side by side and are arranged side by side at the front focal point of a corresponding excitation microlens unit. The fluorescence collecting optical fiber is used to collect the fluorescence signal of the solution to be tested, and outputs the fluorescence signal from the fluorescence output end to the collecting microlens array; The signal-collecting microlens array is used to image different regions of the imaging system.
2. The fluorescence quantitative detection device as described in claim 1, characterized in that, The excitation fiber includes an excitation light input section and an excitation light output section arranged sequentially. The excitation light input section is located near the excitation light input end and is a fiber bundle structure; the excitation light output section is located near the excitation light output end and is a single fiber structure.
3. The fluorescence quantitative detection device as described in claim 1, characterized in that, The fluorescence collecting optical fiber is provided with a fluorescence collecting section, a fluorescence transmission section and a fluorescence output section in sequence. The fluorescence transmission section is an optical fiber bundle structure. The fluorescence collecting section is located near the fluorescence input end, and the fluorescence output section is located near the fluorescence output end. The fluorescence collecting section and the fluorescence output section are both single optical fiber structures.
4. The fluorescence quantitative detection device as described in claim 1, characterized in that, The excitation microlens array includes multiple excitation microlens units arranged in an array, and the signal collection microlens array includes multiple signal collection microlens units arranged in an array. The number of excitation light output terminals, the number of fluorescence input terminals, the number of fluorescence output terminals, the number of excitation microlens units, and the number of signal collection microlens units are all the same as the number of sample quantitative tubes.
5. The fluorescence quantitative detection device as described in claim 1, characterized in that, The front focal point of the excitation microlens unit is located above the sample quantification tube, so that the excitation light enters the test solution directly from above the opening of the sample quantification tube via the excitation microlens unit.
6. The fluorescence quantitative detection device as described in claim 5, characterized in that, The back focal point of the excitation microlens unit is located inside the test solution within the sample quantification tube.
7. The fluorescence quantitative detection device as described in claim 1, characterized in that, The imaging system includes an imaging device and a fluorescence filter. The fluorescence filter is disposed between the fluorescence output end and the imaging device. The imaging device is used to receive the image array output by the signal collection microlens array and to output the image array at one time.
8. The fluorescence quantitative detection device as described in claim 7, characterized in that, The imaging device includes an area array camera or a PMT array photodetector.
9. The fluorescence quantitative detection device as described in claim 1, characterized in that, The focal length range of the excitation microlens unit is 20-30mm.
10. A fluorescence quantitative detection system, characterized in that, Includes the fluorescence quantitative detection device as described in any one of claims 1 to 9.