Device for removing background fluorescence of tissue sample

Abstract

The invention relates to a device for removing background fluorescence of a tissue sample, the device comprises a first shell, a light source module, a sample module, a cooling module and a control module, the light source module and the cooling module are both mounted in an inner cavity of the first shell; the sample module can mix a tissue sample with a reducing agent and then fix the mixture in an inner cavity of the first shell, and the reducing agent comprises a metal hydride reducing agent; the light source module can emit a light source for irradiating the sample module, and the light source and the reducing agent act on the tissue sample to remove background fluorescence of the tissue sample; the cooling module is used for cooling the inner cavity of the first shell; the control module is connected with the cooling module and the light source module. According to the scheme, background fluorescence of a tissue sample can be obviously removed, the background is cleaner after the treated tissue is subjected to antibody staining, and the signal fidelity is improved; the removal effect is thorough, background fluorescence cannot be recovered, and the stability is high; the method basically has no influence on tissue morphology, is basically suitable for all tissues of animals and plants, is not limited by types, and is high in universality.

Description

A device for removing background fluorescence from tissue samples Technical Field

[0001] This invention relates to the technical field of immunoassay devices, and more specifically, to a device for removing background fluorescence from tissue samples. Background Technology

[0002] In fluorescence microscopy of plant and animal tissues, background fluorescence is ubiquitous and has a significant impact. This background fluorescence primarily originates from plant chlorophyll, as well as endogenous substances in animal tissues such as vascular structures, erythrocytes, elastic fibers, collagen fibers, and lipofuscin. Furthermore, cross-linking products formed after tissue fixation also produce characteristic fluorescence. During tissue imaging with fluorescently labeled targets, this background fluorescence significantly interferes with the accurate identification, localization, and quantitative analysis of the target signal, severely limiting the application of high-resolution fluorescence imaging technology in histopathological diagnosis and molecular mechanism research. Currently, strategies for removing tissue background fluorescence are mainly divided into three categories: physical methods, chemical methods, and algorithmic analysis. Physical methods, represented by photobleaching, use high-intensity energy light sources to photodegrade fluorescent substances, thereby reducing the background signal. Algorithmic analysis relies on the spectral splitting technology of the imaging system to separate background fluorescence from the target fluorescence signal. Chemical methods use specific chemical reagents to destroy the molecular structure or optical properties of fluorescent substances, achieving the purpose of removing background fluorescence.

[0003] However, the above methods still face many insurmountable technical bottlenecks in practical applications, exhibiting problems such as poor universality, insufficient stability, and low signal fidelity. Poor universality stems from the need for specific background fluorescence designs tailored to particular tissue types or sources, making them unsuitable for diverse animal and plant tissue samples. Insufficient stability manifests in the tendency for the removed background fluorescence to reappear during secondary antibody labeling or subsequent incubation after photobleaching, leading to background signal rebound. Low signal fidelity arises from the fact that while background light is reduced in samples treated with background light removal reagents, the intensity of the positive signal also decreases, and some reagents severely affect the integrity of tissue morphology. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of existing technologies in removing tissue background fluorescence, such as poor universality, insufficient stability, and low signal fidelity. This invention provides a device for removing background fluorescence from tissue samples. The device in this solution can solve the problems of poor universality, insufficient stability, and low signal fidelity in the current technology for removing tissue background fluorescence.

[0005] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows: A device for removing background fluorescence from tissue samples is provided, comprising a first housing, a light source module, a sample module, a cooling module, and a control module. The light source module and the cooling module are both installed within the cavity of the first housing. The sample module can fix the tissue sample, which is mixed with a reducing agent, within the cavity of the first housing. The reducing agent includes a metal hydride reducing agent. The light source module emits light that irradiates the sample module. After the light source and the reducing agent act on the tissue sample, they remove the background fluorescence. The cooling module is used to cool the cavity of the first housing. The control module is connected to the cooling module and the light source module to control their operation.

[0006] The control module can be wired to the light source module and the cooling module via wires.

[0007] In operation, the device of this invention involves fixing a pre-treated tissue sample and a reducing agent into the inner cavity of the first housing via a sample module. Then, a control module controls the light source module to operate, with the control module controlling the type of tissue sample to direct the light source module to emit light of a corresponding color onto the sample module. Under the combined action of the reducing agent and light, the tissue sample undergoes background fluorescence removal. The tissue sample can be an animal or plant sample, selected according to actual needs. While the light source module is illuminating the sample module, the control module simultaneously controls the cooling module to operate, cooling the inner cavity of the first housing to maintain a suitable temperature.

[0008] The reducing agent is a metal hydride reducing agent, which can change the conjugated structure of the fluorophore, reduce the double bond to a single bond, and reduce the excitation efficiency of the fluorophore. When the light emitted by the light source module shines on the tissue sample, it can excite the target fluorophore, put it in a high-energy state, and accelerate the reduction process.

[0009] This invention provides a device for removing background fluorescence from tissue samples. It can significantly remove background fluorescence from tissue samples, resulting in cleaner backgrounds after antibody staining. The intensity and expression level of antibody-labeled signals are unaffected, and the specific fluorescent labeling signal has a high signal-to-noise ratio, improving signal fidelity. The removal of background fluorescence is thorough, and the background fluorescence will not recover, demonstrating high stability. It has virtually no impact on tissue morphology and is applicable to almost all tissues of plants and animals, regardless of species, exhibiting strong universality.

[0010] Furthermore, the sample module includes a glass slide for holding tissue samples and a reagent chamber for holding reducing agent. The glass slide can be fixed inside the reagent chamber, and the reagent chamber can be fixed inside the first housing. Both the glass slide and the reagent chamber are colorless and transparent to allow light emitted by the light source module to pass through. The glass slide can hold tissue samples, which are then fixed in the reagent chamber after dewaxing and repair on the glass slide. The reagent chamber contains a reducing agent solution to soak the tissue samples on the glass slide.

[0011] In this protocol, tissue samples are sliced, adhered to glass slides, and baked. After baking, the slides are removed, and the tissue samples are dewaxed (slices are sequentially immersed in environmentally friendly dewaxing solution I for 5 min, environmentally friendly dewaxing solution II for 5 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, 85% ethanol aqueous solution (v / v) for 5 min, 75% ethanol aqueous solution (v / v) for 5 min, and then washed with pure water). After dewaxing, the slides undergo retrieval treatment. The specific process is as follows: the dewaxed slides are placed in EDTA retrieval solution at pH 8.0, and then the Seville antigen retrieval instrument is set to a heating temperature of 90℃ for 30 min for retrieval treatment. After cooling to room temperature, the slides are washed three times with PBS for 5 min each time to obtain retrieval treatment slides. The retrieval treatment slides are then immersed in a reagent chamber containing a reducing agent solution.

[0012] While soaking, the sample module was placed inside the first housing cavity and irradiated by the light source module. After irradiation for a period of time, the slide was removed and the tissue sample was washed three times in PBS, 5 minutes each time. After washing, the sample was stained and mounted. The specific procedure was as follows: the nuclei were stained with the nuclear fluorescent dye DAPI, and then mounted with an anti-fluorescence quenching mounting medium. After mounting, the sample was imaged and observed.

[0013] Furthermore, the light source module can emit a first color light and a second color light, where the first color light is white light and the second color light has a wavelength of 475nm to 485nm. The light source module can emit multiple colors of light, further meeting the requirements for background fluorescence removal. Analysis of experimental data shows that when the light emitted by the light source module is white or has a wavelength between 475nm and 485nm, it can basically meet the requirements for background fluorescence removal of all tissue samples. Therefore, the light source module is set to emit white light and light with a wavelength between 475nm and 485nm. The specific color of light used and the specific light intensity value during background fluorescence removal are determined by the control module. The light source module can be a light-emitting panel with a light source capable of emitting both the first and second colors of light, or a light-emitting panel integrating both light sources.

[0014] Furthermore, the cooling module includes a water tank, which is fixedly installed at the top of the inner cavity of the first housing. The water tank's inlet is connected to a cold water source, and its outlet is connected to a water tank. The light source module is attached to the bottom surface of the water tank. The cold water source can be a cold water tap, which is connected to the water tank's inlet via an inlet pipe. A solenoid valve can be installed on the inlet pipe to control its opening and closing. The water tank's outlet can be connected to the water tank via an outlet pipe, which can also be equipped with a solenoid valve to control its opening and closing. This solution uses a water tank to cool the light source module, preventing the heat generated during operation from accumulating on the module and affecting its lifespan. Solenoid valves are installed at both the water tank's inlet and outlet, and these valves are connected to a control module, which controls the valves to flow cold water within the tank. This solution uses water cooling to dissipate heat from the light source module, which not only has high heat dissipation efficiency but also low equipment noise. It eliminates the need to power unnecessary components and reduces the overall power consumption of the device.

[0015] Furthermore, it also includes a temperature sensor for monitoring the temperature of the inner cavity of the first housing. The temperature sensor is fixedly installed in the first housing and connected to the control module. The temperature sensing area can detect the temperature of the inner cavity of the first housing, allowing the tissue sample to undergo background fluorescence removal under suitable temperature conditions. Simultaneously, the temperature sensor can also detect the temperature of the light source module. When the temperature of the light source module reaches a preset high value, it transmits the temperature signal to the control module, which then stops the light source module from operating, preventing it from operating at high temperatures for extended periods and affecting its lifespan.

[0016] Furthermore, the cooling module also includes a fan installed inside the first housing cavity. A vent is provided on the side wall of the first housing. The fan is connected to the control module and is used to accelerate airflow within the first housing cavity. When the temperature sensor detects that the temperature inside the first housing cavity is too high, the control module controls the fan to start operating. The fan's operation accelerates the airflow within the first housing cavity, further speeding up the cooling process.

[0017] Furthermore, the first housing is provided with a pull-out section, which is slidably mounted on the first housing, and the sample module can be fixedly mounted on the pull-out section. The pull-out section is a drawer, which makes it easier to send the sample module into the first housing and facilitates operation by the experimenter. Multiple sample modules can be fixed on the pull-out section at the same time, thereby completing the background fluorescence removal of multiple tissue samples simultaneously and improving efficiency.

[0018] Furthermore, it also includes a second housing, with the control module located within the cavity of the second housing. The second housing separates the control module from the first housing, forming two independent modules for the main body containing the light source module and the control module. The control module can control the module containing the light source module via a wired connection. The arrangement of two independent modules simplifies the structure within the first housing and facilitates the replacement and maintenance of the light source module after prolonged use.

[0019] Furthermore, the control module includes a power supply, a main control signal processing unit, a touch screen, and buttons. The power supply and the main control signal processing unit are fixedly installed in the inner cavity of the second housing. The touch screen is fixedly installed on the surface of the second housing, and the buttons are installed on the surface of the second housing. The main control signal processing unit and the touch screen are both connected to the power supply, and the buttons are connected to the main control signal processing unit. The main control signal processing unit can process data and send commands; the power supply can provide power to the main control signal processing unit, the display screen, the light source module, and the cooling module; the buttons can control the device's on / off state; the experimenter can control the device's operation by operating the touch screen, which can also display relevant experimental data such as temperature. Communication interfaces are provided on the outer surfaces of both the first and second housings. The two communication interfaces are connected by wires to enable the control unit to control the light source module and the cooling module. The light source module and the cooling module inside the first housing are both connected to the communication interface on the first housing; the main control signal processing unit inside the second housing is connected to the communication interface on the second housing.

[0020] Furthermore, a cooling fan is fixedly installed inside the second housing, and the cooling fan is connected to the power supply and the main control signal processing unit. The cooling fan can cool the inside of the second housing.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: The device for removing background fluorescence of tissue samples in this solution can significantly remove the background fluorescence of tissue samples. After antibody staining, the background of the treated tissue is cleaner, the intensity and expression level of antibody labeling signal are not affected, the signal-to-noise ratio of the specific fluorescent labeling signal is high, and the signal fidelity is improved. The removal effect of background fluorescence is thorough, the background fluorescence will not recover, and the stability is high. It has virtually no impact on tissue morphology, is basically applicable to all tissues of animals and plants, is not limited to any species, and has strong universality. Attached Figure Description

[0022] Figure 1 is a schematic diagram of the internal structure of the first housing of a device for removing background fluorescence from tissue samples; Figure 2 is a schematic diagram of the structure of a device for removing background fluorescence from tissue samples; Figure 3 is a schematic diagram of the internal structure of the second housing of a device for removing background fluorescence from tissue samples; Figure 4 shows the imaging results (100×) of different channels of wheat grains after background fluorescence was removed by different methods in Example 4.

[0023] In the attached diagram: 1. First housing; 2. Light source module; 3. Sample module; 4. Temperature sensor; 5. Water tank; 6. Fan; 7. Pull-out section; 8. Second housing; 9. Power supply; 10. Touch screen; 11. Button; 12. Cooling fan; 13. Main control signal processing unit. Detailed Implementation

[0024] The present invention will be further described below with reference to specific embodiments. The accompanying drawings are for illustrative purposes only, representing schematic diagrams rather than actual physical objects, and should not be construed as limiting the scope of this patent. To better illustrate the embodiments of the present invention, some components in the drawings may be omitted, enlarged, or reduced, and do not represent the actual dimensions of the product. It is understandable to those skilled in the art that some well-known structures and their descriptions may be omitted in the drawings.

[0025] In the accompanying drawings of the embodiments of the present invention, the same or similar reference numerals correspond to the same or similar components. In the description of the present invention, it should be understood that if terms such as "upper," "lower," "left," "right," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, they are only for the convenience of describing the present invention 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. Therefore, the terms used to describe positional relationships in the drawings are only for illustrative purposes and should not be construed as limiting the present patent. For those skilled in the art, the specific meaning of the above terms can be understood according to the specific circumstances.

[0026] This embodiment is a first embodiment of a device for removing background fluorescence from tissue samples, as shown in Figures 1-3. It includes a first housing 1, a light source module 2, a sample module 3, a cooling module, and a control module. Both the light source module 2 and the cooling module are installed inside the first housing 1. The sample module 3 can mix the tissue sample with a reducing agent and fix it inside the first housing 1. The reducing agent includes a metal hydride reducing agent. The light source module 2 emits light that irradiates the sample module 3. After the light source and the reducing agent act on the tissue sample, they remove the background fluorescence. The cooling module is used to cool the inside of the first housing 1. The control module is connected to the cooling module and the light source module 2 to control their operation.

[0027] Specifically, metal cyanide reducing agents include metal borohydride reducing agents and metal aluminum hydride reducing agents.

[0028] More preferably, the metal borohydride reducing agent includes one or more of sodium borohydride, sodium cyanoborohydride, sodium triethylborohydride, and lithium borohydride.

[0029] More preferably, the reducing agent for aluminum hydride includes lithium aluminum hydride.

[0030] More preferably, the concentration of the metal hydride reducing agent is 0.05–0.1 M.

[0031] Specifically, the sample module 3 includes a glass slide for holding tissue samples and a reagent chamber for holding reducing agents. The glass slide can be fixed in the inner cavity of the reagent chamber, and the reagent chamber can be fixed in the inner cavity of the first housing 1. Both the glass slide and the reagent chamber are colorless and transparent structures to allow the light source emitted by the light source module 2 to pass through.

[0032] Specifically, the light source module 2 can emit a first color light and a second color light. The first color light is white light, and the wavelength of the second color light is 475nm to 485nm.

[0033] The working principle or process of this embodiment is as follows: When the device of the present invention is in operation, the pre-treated tissue sample and reducing agent are fixedly installed into the inner cavity of the first housing 1 through the sample module 3. Then, the control module controls the light source module 2 to work, and the control module controls the type of tissue sample to control the light source module 2 to emit light of the corresponding color to illuminate the sample module 3. The tissue sample completes the removal of tissue background fluorescence under the combined action of the reducing agent and light. The tissue sample can be an animal or plant sample, which is selected according to actual needs. While the light source module 2 is illuminating the sample module 3, the control module simultaneously controls the cooling module to work, and the cooling module cools the inner cavity of the first housing 1 to maintain the temperature of the inner cavity of the first housing 1 at a suitable temperature for background fluorescence removal.

[0034] The reducing agent is a metal hydride reducing agent. The metal hydride reducing agent can change the conjugated structure of the fluorophore, reduce the double bond to a single bond, and reduce the excitation efficiency of the fluorophore. After the light emitted by the light source module 2 shines on the tissue sample, it can excite the target fluorophore, put it in a high-energy state, and accelerate the reduction process.

[0035] The beneficial effects of this embodiment are as follows: The device for removing background fluorescence from tissue samples in this embodiment can significantly remove the background fluorescence of tissue samples. After antibody staining, the treated tissue has a cleaner background, and the intensity and expression level of antibody labeling signals are not affected. The specific fluorescent labeling signal has a high signal-to-noise ratio, which improves signal fidelity. The background fluorescence removal effect is thorough, and the background fluorescence will not recover, resulting in high stability. It has virtually no impact on tissue morphology and is applicable to almost all tissues of animals and plants, without limitation on species, and has strong universality.

[0036] Example 2 This example is a second embodiment of a device for removing background fluorescence from tissue samples, as shown in Figures 1-3. Based on Example 1, this example further defines the structure of the device.

[0037] Specifically, the cooling module includes a water tank 5, which is fixedly installed at the top of the inner cavity of the first housing 1. The inlet of the water tank 5 is connected to a cold water source, and the outlet of the water tank 5 is connected to a water tank. The light source module 2 is attached to the bottom surface of the water tank 5. The cold water source can be a cold water tap, which is connected to the inlet of the water tank 5 via an inlet pipe. A solenoid valve can be installed on the inlet pipe to control the opening and closing of the inlet pipe. The outlet of the water tank 5 can be connected to the water tank via an outlet pipe, which can also be equipped with a solenoid valve to control the opening and closing of the outlet pipe. The solenoid valve is connected to the control module, enabling the control module to control the operation of the solenoid valve, thereby completing the flow of cold water in the water tank 5.

[0038] Specifically, it also includes a temperature sensor 4 for monitoring the temperature inside the first housing 1. The temperature sensor 4 is fixedly installed in the first housing 1 and is connected to the control module.

[0039] Specifically, the cooling module also includes a fan 6 installed in the inner cavity of the first housing 1. The side wall of the first housing 1 is provided with a vent. The fan 6 is connected to the control module and is used to accelerate the air circulation in the inner cavity of the first housing 1.

[0040] Specifically, the first housing 1 is provided with a pull-out part 7, which is slidably mounted on the first housing 1, and the sample module 3 can be fixedly mounted on the pull-out part 7. The pull-out part 7 is a drawer, and multiple sample modules 3 can be fixed on the pull-out part 7 at the same time.

[0041] The beneficial effects of this embodiment are as follows: Water cooling of the light source module 2 not only provides high heat dissipation efficiency but also reduces equipment noise, eliminates the need to power unnecessary components, and lowers the overall power consumption of the device. The temperature sensing area can detect the temperature inside the first housing 1, allowing tissue samples to undergo background fluorescence removal in a suitable temperature environment. Simultaneously, the temperature sensor 4 can also detect the temperature of the light source module 2. When the temperature of the light source module 2 reaches a preset high value, it transmits the temperature signal to the control module, which then stops the light source module 2 from operating, preventing it from operating at high temperatures for extended periods and affecting its lifespan. The fan 6 accelerates the airflow inside the first housing 1, further speeding up the cooling process. The pull-out section 7 facilitates the insertion of the sample module 3 into the first housing 1, making operation easier for researchers. Multiple sample modules 3 can be simultaneously fixed on the pull-out section 7, allowing for simultaneous background fluorescence removal of multiple tissue samples and improving efficiency.

[0042] Example 3 This example is the third embodiment of a device for removing background fluorescence from tissue samples, as shown in Figures 1-3. Based on Examples 1 and 2, this example further defines the control module.

[0043] Specifically, it also includes a second housing 8, with the control module located inside the second housing 8.

[0044] Specifically, the control module includes a power supply 9, a main control signal processing unit 13, a touch screen display 10, and buttons 11. The power supply 9 and the main control signal processing unit 13 are fixedly installed in the inner cavity of the second housing 8. The touch screen display 10 is fixedly installed on the surface of the second housing 8. The buttons 11 are installed on the surface of the second housing 8. The main control signal processing unit 13 and the touch screen display 10 are both connected to the power supply 9. The buttons 11 are connected to the main control signal processing unit 13.

[0045] Specifically, the outer surfaces of the first housing 1 and the second housing 8 are both provided with communication interfaces. The two communication interfaces are connected by wires to enable the control unit to control the light source module 2 and the cooling module. The light source module 2 and the cooling module inside the first housing 1 are both connected to the communication interface on the first housing 1. The main control signal processing unit 13 inside the second housing 8 is connected to the communication interface on the second housing 8.

[0046] Specifically, a cooling fan 12 is fixedly installed in the inner cavity of the second housing 8, and the cooling fan 12 is connected to the power supply 9 and the main control signal processing unit 13.

[0047] The working principle or process of this embodiment is as follows: the main control signal processing unit 13 can perform data processing and command transmission; the power supply 9 can provide power to the main control signal processing unit 13, the display screen, the light source module 2 and the cooling module; the button 11 can control the switch of the device; the experimenter can control the operation of the device by operating the touch screen 10, and the touch screen 10 can also display relevant experimental data such as temperature.

[0048] The beneficial effects of this embodiment are as follows: By separating the control module from the first housing 1 through the second housing 8, the main body containing the light source module 2 and the control module are formed into two independent mold bases. The arrangement of two independent mold bases makes the internal structure of the first housing 1 simpler and facilitates the replacement and maintenance of the light source module 2 after long-term use of the device. The cooling fan 12 can cool the internal cavity of the second housing 8.

[0049] Example 4 This example is an application example of a device for removing background fluorescence from tissue samples. The device is applied to wheat grains to remove background fluorescence, including the following steps: Step 1: Slicing Take a paraffin block of wheat grains and cut it into 3μm tissue slices using a paraffin slicer. After adhering the slices to a glass slide, bake them in a 65℃ oven for 2 hours, and then remove them for later use.

[0050] Step 2: Dewaxing. The slides are placed in environmentally friendly dewaxing solution I for 5 min, environmentally friendly dewaxing solution II for 5 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, 85% ethanol aqueous solution (v / v) for 5 min, 75% ethanol aqueous solution (v / v) for 5 min, and then washed with pure water.

[0051] Step 3: Repair. Place the dewaxed sections in EDTA repair solution at pH 8.0, then set the heating temperature of the Seville antigen repair instrument to 90℃ and heat the sections for 30 min for repair treatment. After cooling to room temperature, wash three times with PBS for 5 min each time to obtain the repaired sections.

[0052] Step 4: Background Light Removal. The repaired slides were divided into the following 4 groups and treated accordingly: Untreated group: No treatment was performed on the repaired slides; Reagent treatment group: The repaired slides were placed in a reagent chamber containing 0.1M sodium borohydride aqueous solution (i.e., reducing agent) and treated for min. After that, the reagent chamber was removed, and the slides were washed in PBS 3 times for 5 min each time; Light treatment group: The repaired slides were placed in a reagent chamber and irradiated with a 50W white light and a 480nm excitation source for 30 min. After that, the reagent chamber was removed, and the slides were washed in PBS 3 times for 5 min each time; Co-treatment group: The repaired slides were placed in a reagent chamber containing 0.1M sodium borohydride aqueous solution (i.e., reducing agent). Then, the reagent chamber containing the slides was placed in a light source device and irradiated with a 50W white light and a 480nm excitation source for 30 min. After that, the reagent chamber was removed, and the slides were washed in PBS 3 times for 5 min each time.

[0053] Step 5: Nuclear staining and mounting. After staining the nuclei with the nuclear fluorescent dye DAPI staining reagent (brand: Savill Biotechnology, catalog number: G1012), mount the slides with anti-fluorescence quenching mounting medium (brand: Savill, catalog number: G1401).

[0054] Step 6: Imaging was performed using a Nikon upright fluorescence microscope (microscope model: Nikon Eclipse Ci-L, green channel: excitation wavelength 492 nm, emission wavelength 514 nm; red channel: excitation wavelength 555 nm, emission wavelength 569 nm; infrared channel: excitation wavelength 651 nm, emission wavelength 670 nm). With the light source intensity at 100%, images were taken under the following conditions: green channel excitation light, red channel excitation light, and infrared channel excitation light, with an exposure time adjusted to 300 ms; and infrared channel excitation light, with an exposure time adjusted to 1500 ms.

[0055] Experimental results: As shown in Figure 4, the background fluorescence of the ventral groove of wheat grains could only be significantly reduced under the combined treatment of 0.1M sodium borohydride aqueous solution and light system.

[0056] In the specific implementation of the above embodiments, the technical features can be combined in any non-contradictory way. For the sake of brevity, not all possible combinations of the above technical features are described. However, as long as the combination of these technical features is not contradictory, it should be considered to be within the scope of this specification.

[0057] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A device for removing background fluorescence from tissue samples, characterized in that, The system includes a first housing (1), a light source module (2), a sample module (3), a cooling module, and a control module. The light source module (2) and the cooling module are both installed inside the first housing (1). The sample module (3) can mix a tissue sample with a reducing agent and fix it inside the first housing (1). The reducing agent includes a metal hydride reducing agent. The light source module (2) can emit a light source to irradiate the sample module (3). After the light source and the reducing agent act on the tissue sample, they remove the background fluorescence of the tissue sample. The cooling module is used to cool the inside of the first housing (1). The control module is connected to the cooling module and the light source module (2) to control the operation of the light source module (2) and the cooling module.

2. The apparatus for removing background fluorescence from tissue samples according to claim 1, characterized in that, The sample module (3) includes a glass slide for holding tissue samples and a reagent chamber for holding reducing agents. The glass slide can be fixed in the inner cavity of the reagent chamber, and the reagent chamber can be fixed in the inner cavity of the first housing (1). Both the glass slide and the reagent chamber are colorless and transparent structures for the light source emitted by the light source module (2) to pass through.

3. The apparatus for removing background fluorescence from tissue samples according to claim 1, characterized in that, The light source module (2) can emit a first color light and a second color light, wherein the first color light is white light and the wavelength of the second color light is 475nm~485nm.

4. The apparatus for removing background fluorescence from tissue samples according to claim 3, characterized in that, The cooling module includes a water tank (5), which is fixedly installed on the top of the inner cavity of the first housing (1). The water inlet of the water tank (5) is used to connect to a cold water source, and the water outlet of the water tank (5) is used to connect to a water tank. The light source module (2) is attached to the bottom surface of the water tank (5).

5. The apparatus for removing background fluorescence from tissue samples according to claim 1, characterized in that, It also includes a temperature sensor (4) for monitoring the temperature inside the first housing (1), the temperature sensor (4) being fixedly installed in the first housing (1) and connected to the control module.

6. The apparatus for removing background fluorescence from tissue samples according to claim 5, characterized in that, The cooling module also includes a fan (6) installed in the inner cavity of the first housing (1), and the side wall of the first housing (1) is provided with a vent. The fan (6) is connected to the control module.

7. The apparatus for removing background fluorescence from tissue samples according to claim 1, characterized in that, The first housing (1) is provided with a pull-out part (7), which is slidably installed on the first housing (1), and the sample module (3) can be fixedly installed on the pull-out part (7).

8. The apparatus for removing background fluorescence from tissue samples according to claim 1, characterized in that, It also includes a second housing (8), in which the control module is mounted.

9. The apparatus for removing background fluorescence from a tissue sample according to claim 8, characterized in that, The control module includes a power supply (9), a main control signal processing unit (13), a touch screen (10), and buttons (11). The power supply (9) and the main control signal processing unit (13) are fixedly installed in the inner cavity of the second housing (8). The touch screen (10) is fixedly installed on the surface of the second housing (8). The buttons (11) are installed on the surface of the second housing (8). The main control signal processing unit (13) and the touch screen (10) are both connected to the power supply (9). The buttons (11) are connected to the main control signal processing unit (13).

10. The apparatus for removing background fluorescence from a tissue sample according to claim 9, characterized in that, A cooling fan (12) is fixedly installed in the inner cavity of the second housing (8), and the cooling fan (12) is connected to the power supply (9) and the main control signal processing unit (13).

Images