Fluorescent micromanipulation device for preventing quenching of fluorescent sample

Abstract

The invention discloses a fluorescence micromanipulation device for preventing quenching of a fluorescence sample, and relates to the technical field of microscopes, the fluorescence micromanipulation device comprises a stereoscopic microscope body, and the bottom of the stereoscopic microscope body is provided with a height-adjustable base; the bearing hole is formed in the base, and a carrying plate made of a transparent material is mounted in the bearing hole; the anti-quenching operation cabin is detachably mounted on the base, and the cabin wall of the anti-quenching operation cabin is made of a lightproof material and is a hollow frame body; according to the anti-quenching stereoscopic microscope, a controllable operation space with local shading is constructed on the stereoscopic microscope, so that an operator can simultaneously realize effective anti-quenching of a fluorescent sample and sufficient illumination required by fine operation in a bright conventional experiment environment; and the efficiency and the precision of long-time micromanipulation are improved.

Description

Technical Field

[0001] This invention relates to the field of microscopy, and more specifically to a fluorescence micromanipulation device for preventing quenching of fluorescent samples. Background Technology

[0002] In life science research such as neuroscience and developmental biology, it is often necessary to perform immunofluorescence staining on brain slices, embryos, or other tissue sections, followed by delicate operations such as observation, selection, mounting, or dissection under a stereomicroscope. Currently, the white light-illuminated stereomicroscopes commonly used in laboratories have a wide spectral range and contain a large amount of high-energy short-wavelength light.

[0003] However, in existing techniques, when using a conventional white light stereomicroscope to manipulate fluorescent samples, the intense excitation light (mainly the blue-violet component of white light) continuously illuminates the sample, causing photobleaching (quenching) of the fluorescent dye and rapid attenuation of the fluorescence signal. To solve this problem, operators typically need to observe in a completely dark room, relying solely on the limited fluorescence illumination of the microscope itself. However, this method results in insufficient illumination and a dim field of view, significantly impacting the accuracy and efficiency of the operation. This is especially true for delicate actions requiring good background lighting, such as cutting, clamping, and mounting, where the operation is inconvenient and prone to sample damage. Summary of the Invention

[0004] The purpose of this invention is to provide a fluorescence micromanipulation device for preventing quenching of fluorescent samples, so as to solve the technical problem in the prior art that it is difficult to effectively prevent quenching of fluorescent samples and provide sufficient operating illumination in a stereomicroscope under bright ambient light, thereby affecting the efficiency and accuracy of long-term fine operation of fluorescent samples.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a fluorescence micromanipulation device for preventing quenching of fluorescent samples, comprising a stereo microscope body, the bottom of which is equipped with a height-adjustable base;

[0006] A support hole is formed on the base, and a carrier plate made of transparent material is installed in the support hole;

[0007] The anti-quenching operation chamber is detachably installed on the base, and the chamber wall is made of opaque material and is a hollow frame.

[0008] An elastic light-shielding part is installed on the top of the anti-quenching operation chamber;

[0009] A receiving window is provided on the elastic light-shielding part for allowing the objective lens of the stereomicroscope body to extend into the anti-quenching operation chamber;

[0010] A background light source is installed below the base and located below the carrier plate;

[0011] A filter switching assembly is rotatably mounted below the base, and one of the filters is located between the carrier plate and the background light source.

[0012] Furthermore, the vertical cross-sectional profile of the bearing hole is stepped, with the upper diameter being larger than the lower diameter. The carrying plate is installed in the bearing hole with the upper diameter. The top of the base is provided with a mounting groove with a "U" shaped cross-section, and the bottom of the anti-quenching operation chamber is inserted into the mounting groove.

[0013] Furthermore, the outer wall of the anti-quenching operation chamber is provided with a fixedly connected skirt, the bottom of the skirt is in contact with the top of the base, and the inner walls of the anti-quenching operation chamber are coated with a light-absorbing layer.

[0014] Furthermore, the operating hole is provided with a detachable sealing cover plate, the outer wall of the sealing cover plate is provided with rubber strips around it, the front end face of the anti-quenching operating chamber and located on both sides of the operating hole are provided with a magnetic strip one, the sealing cover plate is provided with a magnetic strip two that cooperates with the magnetic strip one, and the front end face of the sealing cover plate is provided with a handle.

[0015] Furthermore, the elastic light-shielding part is made of an opaque material, the outer wall of the base is provided with a downward-extending light-shielding curtain near the bottom, and the bottom of the base is provided with support feet near the four corners.

[0016] Furthermore, the filter switching assembly includes a rotating wheel, a connecting plate, and a retaining ring. The rotating wheel is rotatably connected to the bottom of the base via a pin. The connecting plate is arranged in a ring and fixedly connected to the outer wall of the rotating wheel. The retaining ring is fixedly connected to the other end of the connecting plate, and the filter is embedded in the retaining ring.

[0017] Furthermore, each of the filters is a narrowband filter with a different excitation wavelength of the fluorescent dye.

[0018] Furthermore, each of the retaining rings has a retaining slot on its outer wall, and the base has a positioning plate at its bottom. The positioning plate has a horizontal telescopic rod, which is composed of rods with different diameters at both ends. The two rods are movably connected, with the end of the smaller diameter rod fixedly connected to the positioning plate. A return spring is fitted on the smaller diameter rod, and a retaining block is provided at the front end of the larger diameter rod. The retaining block has a toothed cross-section and cooperates with the retaining slot. The position of the filter is positioned by the retaining block.

[0019] Furthermore, the base has an L-shaped mounting plate at its bottom. The background light source is a switchable wavelength or multi-band light source, which includes at least one long-wavelength emission mode with a peak wavelength greater than 580nm. It is installed at the end of the L-shaped horizontal section of the mounting plate. The bottom of the mounting plate has a power module for supplying power to the background light source.

[0020] Compared with existing technologies, this invention provides a fluorescence micromanipulation device for preventing quenching of fluorescent samples. By integrating the anti-quenching operation chamber, elastic light-shielding section, background light source, and filter switching assembly with a height-adjustable base, it creates a locally controllable optical operation microenvironment. The core advantage of this device is that it allows operators to effectively prevent fluorescence quenching in a normal, bright laboratory environment by physically isolating harmful stray light from the detachable anti-quenching operation chamber. Simultaneously, the use of a spectrally harmless background light source placed below the sample carrier provides sufficient and uniform transmitted illumination to the operation space within the chamber, solving the problems of low operational accuracy, poor efficiency, and fatigue caused by insufficient illumination in dark rooms or low-light conditions. Its integrated structure not only ensures tight light shielding but also facilitates sample handling and the cleaning and maintenance of the device.

[0021] Specifically, the flexible light-shielding section can closely fit the objective lens at different working distances, ensuring the light-shielding seal of the top of the operating chamber. The layout of the background light source and filter switching assembly located below the sample plate allows the switching of excitation light and the provision of harmless operating illumination to both be achieved from below the sample, resulting in a compact and non-interfering optical path. Users can comfortably perform precise microscopic operations in the bright, light-shielded chamber space and quickly select different fluorescence observation channels by rotating the filter switching assembly at the bottom. This enables efficient and convenient switching between "bright-field fine operation" and "dark-field fluorescence observation" on the same device, improving the quality and efficiency of subsequent fluorescent sample processing. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this invention. For those skilled in the art, other drawings can be obtained based on these drawings.

[0023] Figure 1 A schematic diagram of the overall structure of the fluorescence micromanipulation device for preventing quenching of fluorescent samples provided in this embodiment of the invention. Figure 1 ;

[0024] Figure 2 A schematic diagram of the overall structure of the fluorescence micromanipulation device for preventing quenching of fluorescent samples provided in this embodiment of the invention. Figure 2 ;

[0025] Figure 3 This is a schematic diagram of the anti-quenching operation chamber component structure provided in an embodiment of the present invention;

[0026] Figure 4 This is a cross-sectional view of the anti-quenching operating chamber and sealing cover plate, etc., provided in an embodiment of the present invention.

[0027] Figure 5 This is a schematic diagram showing the disassembled structure of the stereomicroscope body and its components, such as the substrate, provided in an embodiment of the present invention.

[0028] Figure 6 This is a schematic diagram of the filter switching assembly and structure provided in an embodiment of the present invention;

[0029] Figure 7 This is a schematic diagram of the structure of components such as retaining rings and retaining blocks provided in an embodiment of the present invention.

[0030] Explanation of reference numerals in the attached figures:

[0031] 1. Stereo microscope body; 2. Base; 3. Mounting hole; 4. Sample plate; 5. Anti-quenching operating chamber; 6. Elastic light-blocking part; 7. Receiving window; 8. Background light source; 9. Filter; 10. Mounting slot; 11. Skirt; 12. Light-absorbing layer; 13. Operating hole; 14. Sealing cover plate; 15. Rubber strip; 16. Magnetic strip one; 17. Magnetic strip two; 18. Handle; 19. Light-blocking curtain; 20. Support foot; 21. Rotary wheel; 22. Connecting plate; 23. Snap ring; 24. Bayonet; 25. Positioning plate; 26. Telescopic rod; 27. Return spring; 28. Mounting plate; 29. ​​Power module; 30. Locking block. Detailed Implementation

[0032] To enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.

[0033] As attached Figure 1 To be continued Figure 7 As shown:

[0034] Example:

[0035] The present invention provides a fluorescence micromanipulation device for preventing quenching of fluorescent samples, including a stereo microscope body 1, the bottom of which is equipped with a height-adjustable base 2.

[0036] A support hole 3 is formed on the base 2, and a carrier plate 4 made of transparent material is installed in the support hole 3;

[0037] The anti-quenching operation chamber 5 is detachably installed on the base 2. The chamber wall of the anti-quenching operation chamber 5 is made of opaque material and is a hollow frame.

[0038] The elastic light-shielding part 6 is installed on the top of the anti-quenching operation chamber 5;

[0039] The accommodation window 7 is opened on the elastic light-shielding part 6 for the objective lens of the stereomicroscope body 1 to extend into the anti-quenching operation chamber 5;

[0040] The background light source 8 is installed below the base 2 and located below the stage 4;

[0041] The filter switching component is rotatably installed below the base 2, and one of the filters 9 is located between the stage 4 and the background light source 8.

[0042] It should be noted that: by integrally integrating the anti-quenching operation chamber 5, the elastic light-shielding part 6, the background light source 8 and the filter switching component with the height-adjustable base 2, a locally controllable optical operation microenvironment is created. The core advantage of this device is that it allows the operator to physically isolate harmful environmental stray light through the detachable anti-quenching operation chamber 5 in an ordinary bright laboratory environment, effectively preventing fluorescence quenching; at the same time, using the background light source 8 placed below the stage 4 and harmless to the spectrum to provide sufficient and uniform transmitted illumination for the operation space inside the chamber, solving the problems of low operation accuracy, poor efficiency and easy fatigue caused by insufficient illumination under darkroom or low-light conditions. Its integrated structure not only ensures the tightness of light shielding, but also facilitates the loading and unloading of samples and the cleaning and maintenance of the device.

[0043] Specifically, the elastic light-shielding part 6 can closely fit the objective lens at different working distances to ensure the light-shielding tightness at the top of the operation chamber. The layout of the background light source 8 and the filter switching component located below the stage 4 enables the switching of excitation light and the provision of harmless operation illumination to be realized from below the sample, with a compact optical path and no interference with each other. The user can calmly perform fine microscopic operations in the bright and light-shielded chamber space, and quickly select different fluorescence observation channels by rotating the filter switching component at the bottom, realizing the efficient and convenient switching of "bright-field fine operation" and "dark-field fluorescence observation" on the same device, improving the quality and efficiency of the subsequent processing of fluorescent samples.

[0044] The stage 4 should be made of a material with high light transmittance (such as optical glass or transparent acrylic) to reduce the attenuation of the two optical paths.

[0045] In this embodiment: the vertical cross-sectional profile of the bearing hole 3 is stepped, with the upper diameter larger than the lower diameter. The stage 4 is installed in the bearing hole 3 with the upper diameter. The top of the base 2 is provided with an installation groove 10 with a cross-section in the shape of a "return". The bottom of the anti-quenching operation chamber 5 is inserted into the installation groove 10.

[0046] It should be noted that the stepped bearing hole 3 design facilitates the installation and positioning of the carrier plate 4. The step formed at the top can reliably support the edge of the carrier plate 4, preventing it from falling. At the same time, this structure allows the top surface of the carrier plate 4 to be basically flush with or slightly lower than the top surface of the base 2, which is convenient for cleaning and does not affect the installation and sealing of the anti-quenching operation chamber 5. The "U"-shaped mounting groove 10 forms a surrounding snap-fit ​​structure, which allows the bottom of the anti-quenching operation chamber 5 to be tightly and vertically inserted and restrained, ensuring the convenience, stability and repeatability of the operation chamber installation, and laying a reliable mechanical foundation for subsequent light-shielding sealing.

[0047] In this embodiment: the outer wall of the anti-quenching operation chamber 5 is provided with a fixedly connected skirt 11, the bottom of the skirt 11 is in contact with the top of the base 2, and the inner walls of the anti-quenching operation chamber 5 are coated with a light-absorbing layer 12.

[0048] It should be noted that the skirt 11 increases the contact area between the anti-quenching operating chamber 5 and the top surface of the base 2, forming a physical barrier that effectively prevents ambient light from entering the operating chamber through the seam between the two, thus enhancing the overall light-shielding effect. The light-absorbing layer 12 (such as matte black paint or black flocked material) coated on the inner wall of the operating chamber can absorb the scattered light generated by the background light source 8 after transmitting the sample, the stray light introduced by the objective lens, and the weak reflected light that may be introduced by the operator's arm, significantly reducing the background light noise inside the chamber. These two measures together create an internal "optical dark field" environment, improving the signal-to-noise ratio of the fluorescence signal, making fluorescence observation clearer, and further reducing potential fluorescence excitation caused by light reflection.

[0049] In this embodiment: the front end face of the anti-quenching operation chamber 5 is provided with an operation hole 13, the operation hole 13 is provided with a detachably connected sealing cover plate 14, the outer wall of the sealing cover plate 14 is provided with rubber strips 15, the front end face of the anti-quenching operation chamber 5 and located on both sides of the operation hole 13 are provided with magnetic strips 16, the sealing cover plate 14 is provided with magnetic strips 17 that cooperate with magnetic strips 16, and the front end face of the sealing cover plate 14 is provided with a handle 18.

[0050] It should be noted that the operating port 13 provides a channel for researchers to insert their hands or tools into the chamber for sample handling. The cooperation of magnetic strips 16 and 17 enables the rapid adsorption and closure of the sealing cover 14. The rubber strip 15 deforms under pressure when the cover is closed, tightly filling the gap between the cover and the chamber wall to form a reliable soft seal, preventing light leakage from the operating port 13. The handle 18 facilitates the application of force to overcome the magnetic force and open the cover. This design, while ensuring light-shielding performance, optimizes the human-machine interface, making sample placement and retrieval, tool replacement, and other operations quick and easy, eliminating the need for repeated disassembly of the entire operating chamber, and improving workflow continuity and efficiency.

[0051] Additionally, it should be noted that the size and shape of the operating hole 13 are not specifically limited here. The size of the operating hole 13 can be rectangular or semi-circular. The opening size of the operating hole 13 should be based on the operator's ability to operate the patch with both hands. A black light-blocking cloth can be added to the top of the operating hole 13 to further improve the light-blocking effect.

[0052] In this embodiment: the elastic light-blocking part 6 is made of an opaque material, the outer wall of the base 2 is provided with a downwardly extending light-blocking curtain 19 near the bottom, and the bottom of the base 2 is provided with support feet 20 near the four corners.

[0053] It should be noted that the elastic light-shielding part 6 is typically made of black sponge, rubber, or flexible pleated fabric, and is arranged around the receiving window 7. When the objective lens is inserted, the elastic material can tightly wrap around the outer wall of the objective lens, adapting to objective lenses of different diameters and the varying insertion depth due to focusing, always maintaining the sealed state of the top opening to prevent light from leaking in from above. The light-shielding curtain 19 (such as a black flexible baffle) extending downward from the outer wall of the base 2 can block ambient light reflected from the experimental table surface or entering from the side and below of the device, further eliminating potential light leakage paths. The support feet 20 raise the distance between the bottom of the base 2 and the table surface, providing space for the swinging of the light-shielding curtain 19, and also facilitating the arrangement and heat dissipation of the filter switching assembly and the background light source 8.

[0054] Specifically, the elastic light-shielding part 6 can be a ring-shaped sponge ring, a silicone sleeve, or a stretchable pleated cover made of multiple layers of black flexible sheets. Its inner diameter is slightly smaller than the outer diameter of a commonly used objective lens, relying on the elasticity of the material itself or the stretchability of the pleated structure to adapt to and hold the objective lens. Preferably, the receiving window 7 is a circular opening with a diameter smaller than the inner diameter of the elastic light-shielding part 6 in its natural state, to ensure that it can maintain basic light protection even when no objective lens is inserted.

[0055] In this embodiment: the filter switching assembly includes a rotating wheel 21, a connecting plate 22 and a retaining ring 23. The rotating wheel 21 is rotatably connected to the bottom of the base 2 by a pin. The connecting plate 22 is arranged in a ring and fixedly connected to the outer wall of the rotating wheel 21. The retaining ring 23 is fixedly connected to the other end of the connecting plate 22. The filter 9 is embedded in the retaining ring 23.

[0056] It should be noted that this mechanical structure achieves compact arrangement and integrated switching of multiple filters 9 within a limited space. The rotating wheel 21, as the core driving component, rotates around a pivot shaft via manual or auxiliary motor drive. The annularly distributed connecting plates 22 synchronously transmit the rotational motion to each retaining ring 23, ensuring that all filters 9 revolve around the same axis. The retaining rings 23 reliably clamp the circular filters 9. By rotating the rotating wheel 21, the user can precisely rotate the desired specific filter 9 to the preset working position directly below the carrier plate 4, making the switching process smooth and intuitive. This component combines the filter 9 storage and working positions into one, resulting in a simple and reliable structure that avoids the cumbersome process and contamination risks associated with individually picking up and installing filters 9 in traditional methods.

[0057] Furthermore, the rotating wheel 21 can extend an operating knob or handle to the side of the base 2 for manual rotation. As a more preferred automation solution, a micro stepper motor can be integrated into the base 2 to drive the rotating wheel 21 to rotate via gears or a belt. The motor is controlled by a controller with buttons corresponding to different filters 9, enabling rapid electric switching and positioning. The telescopic rod 26 positioning mechanism on the positioning plate 25 can be combined with the electronic control scheme, using sensors (such as photoelectric sensors) to detect the position of the bayonet 24, assisting the motor in precise positioning.

[0058] In this embodiment, each of the filters 9 is a narrowband filter 9 with different excitation wavelengths of fluorescent dyes.

[0059] It should be noted that this is crucial for achieving multicolor fluorescence observation and targeted quenching prevention. Each narrowband filter 9 allows only a very narrow range of light waves near its center wavelength (i.e., the characteristic excitation light of a specific fluorescent dye) to pass through with high transmission, while strongly blocking light of other wavelengths. For example, dedicated excitation filters 9 can be configured for dyes such as DAPI, FITC, and TRITC / Cy3. When observing a specific fluorescence, simply switch the corresponding filter 9 into the optical path. The broadband light emitted by the microscope light source is filtered and becomes purified specific excitation light, efficiently exciting the target fluorescence and minimizing the irradiation of the sample by other wavelengths of light (especially short-wavelength harmful light), thereby achieving precise excitation and selective quenching prevention. Switching between different filters 9 can meet the needs of multicolor fluorescence experiments.

[0060] In this embodiment: each of the retaining rings 23 has a retaining slot 24 on its outer wall, and the base 2 has a positioning plate 25 at its bottom. The positioning plate 25 has a horizontal telescopic rod 26. The telescopic rod 26 is composed of rods with different diameters at both ends, and the two rods are movably connected. The end of the smaller diameter rod is fixedly connected to the positioning plate 25. A return spring 27 is sleeved on the smaller diameter rod. A retaining block 30 is provided at the front end of the larger diameter rod. The cross-section of the retaining block 30 is toothed and cooperates with the retaining slot 24. The position of the filter 9 is positioned by the retaining block 30.

[0061] It should be noted that this positioning mechanism ensures the accuracy and repeatability of the working position of the filter 9. When the rotating wheel 21 rotates and the locking slot 24 on a certain locking ring 23 rotates to align with the locking block 30, under the push of the return spring 27, the large-diameter rod drives the toothed locking block 30 forward and embeds it into the locking slot 24. The toothed engagement provides clear mechanical tactile and audible feedback, effectively preventing the filter 9 from shifting in the optical path due to slight external forces or vibrations, ensuring that the excitation light path is always aligned with the center area of ​​the sample, and obtaining a stable and uniform illumination effect. When it is necessary to switch the filter 9, simply pull or rotate the filter 9 assembly gently to overcome the spring force and disengage the locking block 30 from the locking slot 24, and the rotating wheel 21 can continue to rotate. This design improves the reliability and professionalism of the device operation.

[0062] In this embodiment: the bottom of the base 2 is provided with an mounting plate 28 with an "L" shaped vertical cross section, the background light source 8 is a light source that can switch wavelengths or emit multi-band light, and it includes at least one long-wavelength light emission mode with an emission spectrum peak wavelength greater than 580nm, and is installed at the end of the "L" horizontal section of the mounting plate 28. The bottom of the mounting plate 28 is provided with a power module 29 that supplies power to the background light source 8.

[0063] It should be noted that the L-shaped mounting plate 28 supports and suspends the background light source 8 at the optimal illumination position directly below the substrate 4. Its vertical section connects to the base 2 for stable support, while the horizontal section ensures that the light-emitting surface of the light source faces upwards. LEDs with peak wavelengths greater than 580nm (such as amber or red) are chosen as the background light source 8 because the photon energy in this band is too low to excite most common blue, green, and red fluorescent dyes (whose excitation spectra are mainly in the ultraviolet-blue-green region). This provides sufficient illumination within the chamber while avoiding fluorescence quenching caused by the operating illumination itself, achieving "harmless illumination." An independent power module 29 (which may include a battery or power adapter interface) powers the background light source 8, allowing its brightness to be adjusted independently of the microscope's main light source, thus enhancing operational flexibility.

[0064] Preferably, in practical applications, the background light source 8 can also be a tunable wavelength LED or a multi-band light source containing a short wavelength emission unit (e.g., a 488nm blue filter for exciting GFP), and can be switched between the operating mode and the fluorescence excitation mode by the control circuit to adapt to the needs of different fluorescent dyes.

[0065] Specifically, the power module 29 can integrate a brightness adjustment circuit (such as a potentiometer or PWM dimming circuit), allowing the operator to flexibly adjust the brightness of the background light according to the sample thickness, personal visual habits, or different operating stages (such as high brightness when searching for samples and soft light when performing delicate operations). The switch of the background light source 8 can be located on the side of the base 2 in an easily accessible position, or managed by a control circuit linked to the filter switching assembly.

[0066] The operating procedure of this invention is as follows: When using this device, the conventional operating procedure is as follows: In a bright room, open the sealing cover 14, place the sample in, and close it; turn on the background light source 8, and perform fine operations such as positioning, clamping, and trimming of the sample under uniform long-wavelength transmitted light through the eyepiece; when it is necessary to check the fluorescence, keep the background light source 8 on (because it is harmless), and switch the filter 9 to the target channel. The light emitted by the microscope light source is purified by the filter 9 and then excites the fluorescence. At this time, a bright fluorescence signal can be observed under a dark background (the light absorption environment inside the chamber); after observation, switch the filter 9 away from the excitation position or turn off the microscope light source, and continue to operate under the background light.

[0067] The foregoing has only described certain exemplary embodiments of the present invention by way of illustration. Undoubtedly, those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the foregoing drawings and descriptions are illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.

Claims

1. A fluorescence micromanipulation device for preventing quenching of fluorescent samples, characterized in that, Comprising: A stereomicroscope body (1), the bottom of which is equipped with a height-adjustable base (2); A carrying hole (3) is opened on the base (2), and a stage plate (4) made of a transparent material is installed in the carrying hole (3); An anti-quenching operation chamber (5) is detachably installed on the base (2), and the chamber wall of the anti-quenching operation chamber (5) is made of a light-impervious material and is a hollow frame; An elastic light-shielding part (6) is installed on the top of the anti-quenching operation chamber (5); An accommodation window (7) is opened on the elastic light-shielding part (6) for the objective lens of the stereomicroscope body (1) to extend into the anti-quenching operation chamber (5); A background light source (8) is installed below the base (2) and is located below the stage plate (4); A filter switching component is rotatably installed below the base (2), and one of the filters (9) is located between the stage plate (4) and the background light source (8).

2. The fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 1, characterized in that, The vertical cross-sectional profile of the carrying hole (3) is stepped, with the upper diameter being larger than the lower diameter. The stage plate (4) is installed in the carrying hole (3) with the upper diameter. The top of the base (2) is provided with an installation groove (10) with a "return" - shaped cross-section, and the bottom of the anti-quenching operation chamber (5) is inserted into the installation groove (10).

3. A fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 2, characterized in that, The outer wall of the anti-quenching operation chamber (5) is provided with a fixedly connected skirt (11), the bottom of the skirt (11) is mutually adhered to the top of the base (2), and the inner walls around the anti-quenching operation chamber (5) are coated with an absorbent layer (12).

4. A fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 1, characterized in that, An operation hole (13) is opened on the front end face of the anti-quenching operation chamber (5), and a detachably connected sealing cover plate (14) is provided on the operation hole (13). Rubber strips (15) are provided around the outer wall of the sealing cover plate (14). Magnetic attraction strips one (16) are provided on both sides of the operation hole (13) on the front end face of the anti-quenching operation chamber (5). Magnetic attraction strips two (17) that cooperate with the magnetic attraction strips one (16) are provided on the sealing cover plate (14), and a handle (18) is provided on the front end face of the sealing cover plate (14).

5. A fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 1, characterized in that, 6. A fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 1, characterized in that, The elastic light-shielding part (6) is made of a light-impervious material. A light-shielding curtain (19) extending downward is provided at a position near the bottom of the outer wall of the base (2), and support feet (20) are provided at positions near the four corners of the bottom of the base (2).

7. A fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 6, characterized in that, The filter switching component includes a rotating wheel (21), a connecting plate (22) and a clamping ring (23). The rotating wheel (21) is rotationally connected to the bottom of the base (2) through a pin shaft. The connecting plates (22) are annularly distributed and fixedly connected to the outer wall of the rotating wheel (21). The clamping ring (23) is fixedly connected to the other end of the connecting plate (22), and the filter (9) is embedded in the clamping ring (23). Each of the filters (9) is a narrow-band filter (9) with different fluorescence dye characteristic excitation wavelengths.

8. A fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 7, characterized in that, Each of the retaining rings (23) has a retaining slot (24) on its outer wall. The base (2) has a positioning plate (25) at its bottom. The positioning plate (25) has a horizontal telescopic rod (26). The telescopic rod (26) is composed of rods with different diameters at both ends. The two rods are movably connected. The end of the smaller diameter rod is fixedly connected to the positioning plate (25). A return spring (27) is sleeved on the smaller diameter rod. A retaining block (30) is provided at the front end of the larger diameter rod. The cross-section of the retaining block (30) is toothed and cooperates with the retaining slot (24). The position of the filter (9) is positioned by the retaining block (30).

9. A fluorescence micromanipulation device for preventing quenching of fluorescent samples according to claim 1, characterized in that, The base (2) has an L-shaped mounting plate (28) at the bottom. The background light source (8) is a light source that can switch wavelengths or emit multi-band light. It includes at least one long-wavelength light emission mode with a peak wavelength of emission spectrum greater than 580nm and is installed at the end of the L-shaped horizontal section of the mounting plate (28). The bottom of the mounting plate (28) is provided with a power supply module (29) that supplies power to the background light source (8).

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