Fluorescence detection device

Abstract

The invention discloses a fluorescence detection device which is used for collecting fluorescence signals on a sample film, the fluorescence detection device comprises an excitation light source, a detection element and a camera obscura, the excitation light source and the detection element are both arranged in the camera obscura, the excitation light source acts on the sample film to excite the sample film to generate the fluorescence signals, the detection element is located below the sample film, and the detection element is located below the sample film. The detector is used for detecting a fluorescence signal and converting the fluorescence signal into an electric signal; the fluorescence detection device further comprises a first filtering element, and the first filtering element is arranged above the sample film, is positioned below the excitation light source and is used for filtering interference light emitted by the excitation light source; and / or the fluorescence detection device further comprises a second filter element, the second filter element is arranged below the sample membrane, and the second filter element is located above the detection element and used for filtering interference light reaching the detection element; wherein the second filter element is a micro optical fiber plate.

Description

Technical Field

[0001] The present invention relates to a fluorescence detection device. Background Art

[0002] Fluorescent labeling is widely used in the detection of markers in biological samples. By collecting the fluorescence signals generated by the samples, information such as the distribution or content of relevant labeled substances can be detected. It irradiates the fluorescently labeled biological samples with excitation light to generate fluorescence signals, which are then detected by a detection element and transmitted to a computer, displayed in the form of a color image and quantitatively analyzed.

[0003] The detection of traditional fluorescently labeled sample signals mainly uses the optical imaging method of a digital camera and an optical lens for fluorescence signal collection and analysis, mainly consisting of main components such as a digital camera, an optical lens, an excitation light module, an excitation light source filter, a reflected light filter, etc. In the traditional optical imaging method, due to the large distance between the digital camera and the lens and the biological sample, the digital camera can only collect a very small part of the fluorescence signals emitted by the sample, so there are certain limitations in the detection of weak light signals. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to overcome the defect that in the prior art, the digital camera can only collect a very small part of the fluorescence signals emitted by the sample, resulting in certain limitations in the detection of weak light signals, and to provide a fluorescence detection device.

[0005] The present invention solves the above technical problem through the following technical solutions:

[0006] The present invention discloses a fluorescence detection device for collecting fluorescence signals on a sample film. The fluorescence detection device includes an excitation light source, a detection element, and a dark box. The excitation light source and the detection element are both arranged in the dark box. The excitation light source acts on the sample film to excite fluorescence signals on the sample film. The detection element is located below the sample film and is used to detect the fluorescence signals and convert the fluorescence signals into electrical signals;

[0007] The fluorescence detection device further includes a first filtering element, which is arranged above the sample film and below the excitation light source, and is used to filter the interfering light emitted by the excitation light source;

[0008] And / or, the fluorescence detection device further includes a second filtering element, which is arranged below the sample film and above the detection element, and is used to filter the interfering light reaching the detection element; wherein the second filtering element is a microfiber plate.

[0009] In this solution, adopting the above structural form, by setting up a light-tight box, it provides a light-shielding environment for the excitation light source to excite the sample film to generate a silver light signal and for the detection element to detect the influence signal, effectively avoiding interference from external light and ensuring the accuracy of the detection result. The excitation light source is located above the sample film, so that the fluorescence substance on the sample film can be excited by the excitation light source to generate a signal. The detection element is located below the sample film and is set at a close distance, enabling the detection element to capture the fluorescence signal emitted by the sample film, reducing the loss of the signal during transmission, and improving the detection efficiency and accuracy. The first filtering element is arranged below the laser element and above the sample film, and can filter the interfering light emitted by the excitation light source to avoid the adverse effects of stray light on the fluorescence signal. The second filtering element is a microfiber plate, which is located above the detection element and below the sample film, and performs secondary filtering before the light is transmitted to the detection element to further eliminate the remaining interfering light. With the special optical conduction and filtering characteristics of the microfiber plate, it can not only screen out the fluorescence band that meets the detection requirements, but also enhance the conduction stability of the light signal, making the fluorescence signal finally received by the detection element pure and stable, thus greatly improving the detection accuracy and reliability of the entire fluorescence detection device.

[0010] Preferably, the thickness of the microfiber plate is 1 mm - 5 mm;

[0011] And / or, the fiber diameter of the microfiber plate is 3 microns - 20 microns.

[0012] In this solution, adopting the above structural form enables the microfiber plate to effectively control the scattering and loss of light during transmission while maintaining its own structural stability, and improves the intensity of the fluorescence signal transmitted to the detection element;

[0013] The light diameter of the microfiber plate is 3 microns - 20 microns, which can improve the spatial resolution, enabling the detection element to more accurately detect the change of the fluorescence signal in a tiny area on the sample film.

[0014] Preferably, the detection element includes a CMOS detector.

[0015] Preferably, the size of the CMOS detector is 3 cm × 3 cm - 15 cm × 15 cm;

[0016] And / or, the pixel size of the CMOS detector is 20 microns × 20 microns - 100 microns × 100 microns.

[0017] Preferably, the fluorescence detection device further includes a beam expander, which is arranged between the sample film and the first filtering element and is used to diffuse the light filtered by the first filtering element into a surface light source.

[0018] In this solution, adopting the above structural form, the beam expander is located between the sample film and the first filtering element, and can diffuse the light filtered by the first filtering element to convert it into a uniform surface light source, thereby effectively solving the problem of uneven light spots, ensuring that each area on the sample film can receive the excitation light evenly, and thus improving the excitation efficiency and uniformity of the fluorescence signal. The uniform surface light source helps to reduce signal fluctuations and errors during the detection process, enabling the detection element to capture the fluorescence signal more stably and accurately, and enhancing the reliability and repeatability of the entire detection device. In addition, the application of the beam expander can also expand the coverage range of the excitation light, ensuring that when performing fluorescence detection on a sample film with a large area, the signal intensity of each detection point is still consistent, which can significantly improve the detection efficiency and the accuracy of the data. At the same time, this uniform diffusion treatment of light also helps to optimize the working conditions of the detection element, avoiding problems such as saturation or weak signal of the detection element caused by excessive or insufficient local light intensity, and further enhancing the detection sensitivity and dynamic range.

[0019] Preferably, the diffusion angle of the light after passing through the beam expander is 5° - 30°.

[0020] Preferably, the outer edge of the beam expander coincides with the outer edge of the light-sensitive area of the detection element;

[0021] Or, the outer edge of the beam expander is located outside the outer edge of the light-sensitive area of the detection element.

[0022] In this solution, adopting the above structural form, when the outer edge of the beam expander coincides with the outer edge of the light-sensitive area of the detection element, the expanded surface light source can accurately match the light-sensitive area of the detection element, ensuring that the entire light-sensitive area evenly receives the filtered and diffused excitation light, avoiding light overflow or deficiency, enabling the detection element to efficiently capture the fluorescence signal generated on the sample film, and improving the detection accuracy and stability. When the outer edge of the beam expander is located outside the outer edge of the light-sensitive area of the detection element, it can further expand the coverage range of the excitation light, ensuring that even in the edge area of the light-sensitive area of the detection element, sufficient intensity and uniform excitation light can be received, which is particularly beneficial for the case where the distribution of fluorescent substances on the sample film is uneven or the light-sensitive area of the detection element is slightly smaller than the beam expander, and can effectively reduce the detection error caused by uneven light intensity, and enhance the detection uniformity and reliability.

[0023] Preferably, the fluorescence detection device further includes a condenser, and the condenser is arranged between the excitation light source and the first filtering element for converging the excitation light source into a light beam.

[0024] In this solution, adopting the above structural form enables the light emitted by the excitation light source to better pass through the first filtering element, so that the interference light can be filtered by the first filtering element, reducing the influence of the interference light on the sample film.

[0025] Preferably, the second filtering element is one or more of a single-channel filter, a single-channel band-pass filter, and a multi-position filter lens.

[0026] Preferably, the excitation light source is one or more of an LED lamp, a deuterium lamp, a halogen lamp, a solid-state laser emitter, or a gaseous laser emitter.

[0027] The positive and progressive effects of the present invention are as follows:

[0028] By providing a dark box, it provides a light-shielding environment for the excitation light source to excite the sample film to generate a silver light signal and for the detection element to detect the influence signal, effectively avoiding interference from external light and ensuring the accuracy of the detection result. The excitation light source is located above the sample film, so that the fluorescence substance on the sample film can be excited by the excitation light source to generate a signal. The detection element is located below the sample film and is set at a close distance, so that the detection element can capture the fluorescence signal emitted by the sample film, reducing the loss of the signal during transmission and improving the detection efficiency and accuracy. The first filtering element is arranged below the laser element and above the sample film, and can filter the interference light emitted by the excitation light source, avoiding the adverse influence of stray light on the fluorescence signal. The second filtering element is a microfiber plate, which is located above the detection element and below the sample film, and performs secondary filtering before the light is transmitted to the detection element, further eliminating the residual interference light. With the special optical conduction and filtering characteristics of the microfiber plate, it can not only screen out the fluorescence band that meets the detection requirements, but also enhance the conduction stability of the light signal, making the fluorescence signal finally received by the detection element pure and stable, thus greatly improving the detection accuracy and reliability of the entire fluorescence detection device. Description of the Drawings

[0029] Figure 1 It is a schematic structural diagram of the fluorescence detection device according to an embodiment of the present invention.

[0030] Figure 2 It is a schematic cross-sectional structural diagram of the fluorescence detection device according to an embodiment of the present invention.

[0031] Figure 3 It is a partial cross-sectional structural diagram of the fluorescence detection device according to an embodiment of the present invention.

[0032] Figure 4 It is a partial cross-sectional schematic diagram of the fluorescence detection device according to an embodiment of the present invention.

[0033] Description of the Reference Numerals:

[0034] Fluorescence detection device 100

[0035] Laser power supply 1

[0036] Detection element 2

[0037] Dark box 3

[0038] First filtering element 4

[0039] Second filtering element 5

[0040] Beam expander element 6

[0041] Condensing element 7

[0042] Sample stage 8

[0043] Detection area 81 Detailed implementation manner

[0044] The present invention will be further described below by way of embodiments, but the present invention is not limited to the scope of the described embodiments thereby.

[0045] As Figures 1 to 4 shown, this embodiment provides a fluorescence detection device for collecting fluorescence signals on a sample film. The fluorescence detection device includes an excitation light source, a detection element, and a dark box. The excitation light source and the detection element are both disposed inside the dark box. The excitation light source acts on the sample film to excite fluorescence signals on the sample film. The detection element is located below the sample film and is used to detect the fluorescence signals and convert the fluorescence signals into electrical signals. The silver fluorescence detection device further includes a first filtering element and a second filtering element. For the first filtering element and the second filtering element, there can be the following several implementation manners. The first implementation manner is that the first filtering element is disposed above the sample film and is located below the excitation light source for filtering the interfering light emitted by the excitation light source. The second implementation manner is that the second filtering element is disposed below the sample film and is located above the detection element for filtering the interfering light reaching the detection element. Among them, the second filtering element is a micro optical fiber plate. The third implementation manner is that the first filtering element is disposed above the sample film and is located below the excitation light source for filtering the interfering light emitted by the excitation light source, and the second filtering element is disposed below the sample film and is located above the detection element for filtering the interfering light reaching the detection element. Among them, the second filtering element is a micro optical fiber plate.

[0046] Preferably, the first filtering element is disposed above the sample film and below the excitation light source for filtering the interfering light emitted by the excitation light source, and the second filtering element is disposed below the sample film and above the detection element for filtering the interfering light reaching the detection element; wherein the second filtering element is a micro optical fiber plate. With the above structural form, by setting up a light-tight box, a light-shielding environment is provided for the excitation light source to excite the silver light signal of the sample film and the detection element to detect the influence signal, effectively avoiding the interference of external light and ensuring the accuracy of the detection result. The excitation light source is located above the sample film, so that the fluorescent substance on the sample film can be excited by the excitation light source to generate a signal. The detection element is located below the sample film and is set at a close distance, so that the detection element can capture the fluorescent signal emitted by the sample film, reducing the loss of the signal during transmission and improving the detection efficiency and accuracy. The first filtering element is disposed below the laser element and above the sample film, and can filter the interfering light emitted by the excitation light source to avoid the adverse effect of stray light on the fluorescent signal. The second filtering element is a micro optical fiber plate, located above the detection element and below the sample film, and performs secondary filtering before the light is transmitted to the detection element, further eliminating the residual interfering light. With the special optical conduction and filtering characteristics of the micro optical fiber plate, it can not only screen out the fluorescent band that meets the detection requirements, but also enhance the conduction stability of the optical signal, making the fluorescent signal finally received by the detection element pure and stable, thus greatly improving the detection accuracy and reliability of the entire fluorescence detection device.

[0047] Specifically, the above-mentioned "above" refers to the direction opposite to the illumination direction of the excitation light source; the above-mentioned "below" refers to the illumination direction of the excitation light source.

[0048] In specific use, the micro optical fiber plate is a specific wavelength filter after multi-layer sputtering coating treatment; in other embodiments, the micro optical fiber plate can also be a commercially available finished product.

[0049] In addition, as Figure 2 and Figure 4 shown, the fluorescence detection device further includes a sample stage, and the biological sample film is placed in the detection area of the sample stage. After the light passes through the diffusion element, it can at least cover the detection area.

[0050] In the detection of biological membranes, a light-tight box refers to a specially designed enclosed light-shielding device or experimental environment, whose core function is to ensure the stability of photosensitive reactions or optical signals during the detection process by completely isolating external light interference. The light-tight box is usually composed of light-impermeable materials (such as black metal, light-shielding coated plastic or airtight light-shielding cloth).

[0051] For the micro - optical fiber plate, its thickness can be 1 mm - 5 mm; adopting the above - mentioned structural form enables the micro - optical fiber plate to effectively control the scattering and loss of light during transmission while maintaining its own structural stability, and improves the intensity of the fluorescence signal transmitted to the detection element. In other embodiments, the thickness of the micro - optical fiber plate can be adjusted according to actual needs and is not limited herein.

[0052] The fiber diameter of the micro - optical fiber plate is 3 microns - 20 microns, which can improve the spatial resolution, enabling the detection element to more precisely detect the fluorescence signal changes in the tiny areas on the sample film. In other embodiments, the fiber diameter of the micro - optical fiber plate can be adjusted according to actual needs and is not limited herein.

[0053] In this embodiment, the detection element includes a CMOS detector. In other embodiments, the detection element can also adopt other forms and is not limited herein.

[0054] For the CMOS detector, its size can be 3 cm × 3 cm - 15 cm × 15 cm, and the pixel size is 20 microns × 20 microns - 100 microns × 100 microns.

[0055] As Figure 2 and Figure 3 shown, the fluorescence detection device further includes a beam - expanding element. The beam - expanding element is arranged between the sample film and the first filtering element and is used to diffuse the light filtered by the first filtering element into a surface light source. Adopting the above - mentioned structural form, the beam - expanding element is located between the sample film and the first filtering element, and can diffuse the light filtered by the first filtering element, converting it into a uniform surface light source. Thus, it can effectively solve the problem of uneven light spots, ensure that each area on the sample film can receive the excitation light evenly, thereby improving the excitation efficiency and uniformity of the fluorescence signal. The uniform surface light source helps to reduce the signal fluctuations and errors during the detection process, enabling the detection element to capture the fluorescence signal more stably and accurately, and enhancing the reliability and repeatability of the entire detection device. In addition, the application of the beam - expanding element can also expand the coverage range of the excitation light, ensuring that the signal intensity of each detection point is consistent when performing fluorescence detection on a sample film with a large area, which can significantly improve the detection efficiency and the accuracy of the data. At the same time, this uniform diffusion process of light also helps to optimize the working conditions of the detection element, avoiding problems such as saturation or weak signal of the detection element caused by excessive or insufficient local light intensity, and further enhancing the sensitivity and dynamic range of the detection.

[0056] In this embodiment, the diffusion angle of the light after passing through the beam - expanding element is 5° - 30°. In other embodiments, the diffusion angle of the light can be adjusted according to actual needs and is not limited herein.

[0057] The outer edge of the beam expander element coincides with the outer edge of the light-sensitive area of the detection element; or, the outer edge of the beam expander element is located outside the outer edge of the light-sensitive area of the detection element. With the above structural form, when the outer edge of the beam expander element coincides with the outer edge of the light-sensitive area of the detection element, the expanded surface light source can accurately match the light-sensitive area of the detection element, ensuring that the entire light-sensitive area evenly receives the filtered and diffused excitation light, avoiding light overflow or deficiency, enabling the detection element to efficiently capture the fluorescence signal generated on the sample film, and improving the accuracy and stability of the detection. When the outer edge of the beam expander element is located outside the outer edge of the light-sensitive area of the detection element, it can further expand the coverage range of the excitation light, ensuring that even in the edge area of the light-sensitive area of the detection element, sufficient intensity and uniform excitation light can be received, which is particularly beneficial for the case where the fluorescence substances on the sample film are unevenly distributed or the light-sensitive area of the detection element is slightly smaller than the beam expander element, effectively reducing the detection error caused by uneven light intensity and enhancing the uniformity and reliability of the detection.

[0058] As Figure 2 and Figure 3 shown, the fluorescence detection device further includes a condenser element, which is arranged between the excitation light source and the first filtering element and is used to converge the excitation light source into a light beam. With the above structural form, the light emitted by the excitation light source can better pass through the first filtering element, so that the interference light can be filtered by the first filtering element, reducing the influence of the interference light on the sample film.

[0059] In this embodiment, the second filtering element is one or more of a single-channel filter, a single-channel band-pass filter, and a multi-bit filter lens.

[0060] The excitation light source is one or more of an LED lamp, a deuterium lamp, a halogen lamp, a solid-state laser emitter, or a gaseous laser emitter.

[0061] Although the specific embodiments of the present invention have been described above, those skilled in the art should understand that this is only an example, and the protection scope of the present invention is defined by the appended claims. Without departing from the principles and essence of the present invention, those skilled in the art can make various changes or modifications to these embodiments, but these changes and modifications all fall within the protection scope of the present invention.

Claims

1. A fluorescence detection device for collecting fluorescence signals on a sample membrane, characterized in that: The fluorescence detection device comprises an excitation light source, a detection element and a dark box, wherein the excitation light source and the detection element are both arranged in the dark box, the excitation light source acts on the sample film to excite the sample film to generate a fluorescence signal, and the detection element is located below the sample film and is used to detect the fluorescence signal and convert the fluorescence signal into an electrical signal; The fluorescence detection device further includes a first filter element, which is disposed above the sample membrane and below the excitation light source, and is used to filter interference light emitted by the excitation light source; And / or, the fluorescence detection device also includes a second filter element, which is arranged below the sample membrane and above the detection element, and is used to filter interference light reaching the detection element; wherein the second filter element is a micro fiber optic plate.

2. The fluorescence detection device according to claim 1, characterized in that: The thickness of the micro optical fiber plate is 1mm-5mm; And / or, the diameter of the optical fiber of the micro optical fiber plate is 3 microns to 20 microns.

3. The fluorescence detection device according to claim 1, characterized in that: The detection element includes a CMOS detector.

4. The fluorescence detection device according to claim 3, characterized in that: The size of the CMOS detector is 3cm×3cm-15cm×15cm; And / or, the pixel size of the CMOS detector is 20 microns×20 microns-100 microns×100 microns.

5. The fluorescence detection device according to claim 1, characterized in that: The fluorescence detection device further comprises a beam expansion element, which is disposed between the sample membrane and the first filter element and is used to diffuse the light filtered by the first filter element into a surface light source.

6. The fluorescence detection device according to claim 5, characterized in that: The diffusion angle of the light after passing through the beam expander is 5°-30°.

7. The fluorescence detection device according to claim 5, characterized in that: The outer edge of the beam expansion element coincides with the outer edge of the light-sensing area of ​​the detection element; Alternatively, the outer edge of the beam expanding element is located outside the outer edge of the photosensitive area of ​​the detection element.

8. The fluorescence detection device according to claim 1, characterized in that: The fluorescence detection device further comprises a focusing element, which is disposed between the excitation light source and the first filter element and is used to focus the excitation light source into a light beam.

9. The fluorescence detection device according to claim 1, characterized in that: The second filter element is one or more of a single-channel filter, a single-channel bandpass filter, and a multi-position filter.

10. The fluorescence detection device according to any one of claims 1 to 9, characterized in that: The excitation light source is one or more of an LED lamp, a deuterium lamp, a halogen lamp, a solid-state laser emitter or a gaseous laser emitter.

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