A device and method for batch staining of electron microscopy ultrathin section samples
By designing a batch staining device for ultrathin electron microscope sections, and employing a copper mesh box snap-fit structure and a dual staining step, the problems of easy damage to the mesh and time-consuming operation were solved, achieving efficient and low-cost staining results.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SOUTH CHINA AGRICULTURAL UNIVERSITY
- Filing Date
- 2026-04-27
- Publication Date
- 2026-06-09
AI Technical Summary
Existing methods for staining ultrathin sections of electron microscope samples have problems such as easy deformation or damage of the mesh, time-consuming and labor-intensive operation, and difficulty in controlling staining quality.
A batch staining device for ultrathin electron microscope sections was designed, including a tray, a tray cover, a copper mesh box, a handle, and a wax tray. The mesh is fixed by a snap-fit structure between the bottom box and the lid of the copper mesh box. Combined with a dual staining step of uranium acetate and lead citrate, the device achieves stable clamping and efficient staining of the mesh.
It effectively prevents deformation and damage to the dyeing net, improves dyeing efficiency, ensures dyeing quality, and reduces operating costs.
Smart Images
Figure CN122171296A_ABST
Abstract
Description
Technical Field
[0001] This invention application relates to the field of transmission electron microscopy sample pretreatment tools, specifically to a batch staining device and method for ultrathin section samples for electron microscopy. Background Technology
[0002] Observing the microscopic morphology of biological samples using transmission electron microscopy (TEM) requires a series of preparation steps, followed by slicing the biological sample into ultrathin sections approximately 70 nanometers thick and attaching them to a grid approximately 3 millimeters in diameter. The ultrathin sections on the grid are then stained with heavy metals, typically using a double staining of uranium acetate and lead citrate. Currently, three common staining methods exist: One method involves placing a drop of uranium acetate staining solution onto a wax tray or sealing film, inverting the grid onto the droplet, and allowing the ultrathin section to contact the staining solution. After staining for approximately 30 minutes, the grid is removed with tweezers, rinsed several times with distilled water, and then blotted dry with filter paper. The same procedure is then repeated using lead citrate staining solution. Note that when using lead citrate, sodium hydroxide should be added nearby to prevent contamination from airborne carbon dioxide and other pollutants. After the grid has dried, electron microscopy observation is performed. This method has three problems: first, it requires repeatedly holding the screen with tweezers, which can easily deform the screen or damage the ultrathin section; second, manually holding the screen for staining and multiple cleanings is time-consuming, labor-intensive, and inefficient; and third, the staining and cleaning times are difficult to synchronize, making it hard to guarantee staining quality. Another method involves using elastic materials such as silicone or tooth wax to create staining plates or strips, with appropriately sized and deep slits to hold the screen, and then placing the staining plate or strip in a staining tube or box for staining and cleaning. The problem with this method is that when the slits of the staining plate or strip hold the screen, it can easily cause the screen to fall off or damage the supporting membrane on the screen. The third method involves more complex staining equipment, but this method requires more complex equipment and is more expensive. Summary of the Invention
[0003] To address the aforementioned issues, a batch staining device and method for electron microscopy ultrathin section samples are provided. By setting up a tray, tray cover, copper mesh box, handle, and wax tray, the mesh carrier can be repeatedly handled with tweezers without needing to be repeatedly gripped, effectively preventing mesh carrier deformation and damage to the ultrathin sections.
[0004] To address the problems of the existing technology, this invention application provides the following technical solution: A batch staining device for electron microscopy ultrathin section samples includes: tray; A wax tray is placed on the tray, the wax tray is open to the upward and has grooves formed on it; A copper mesh box is used to hold multiple carrier meshes, each carrier mesh containing an ultrathin slice to be stained, and the copper mesh box can be placed into the groove. A handle is provided on the copper mesh box for picking up and putting down the copper mesh box; A tray cover, opaque, used to cover a tray.
[0005] Preferably, the copper mesh box includes a bottom box and a lid; The bottom box has multiple placement slots. When viewed from above, the placement slots are rhomboid in shape. The placement slots are used to place the carrier net. The carrier net can be placed vertically in the placement slots, and the carrier net is just caught in the two opposite angles of the rhomboid shape of the placement slot. The handle is fixedly connected to the lid of the copper mesh box.
[0006] Preferably, a first through hole is provided at the bottom of each placement slot, the first through hole extends vertically through the bottom box, and the diameter of the first through hole is smaller than the diameter of the carrier net; A second perforation is provided on the box cover at the position corresponding to each placement slot, and the diameter of the second perforation is smaller than the diameter of the carrier net.
[0007] Preferably, the bottom box is snapped onto the lid.
[0008] Preferably, a locking strip is provided on both opposite sides of the lid and below the lid, and a locking groove is formed between the locking strip and the lid; an insertion part is provided on both sides of the corresponding locking groove of the bottom box, and the insertion part can be inserted into the locking groove to realize the locking between the bottom box and the lid. When the bottom box is inserted into the lid, the placement groove, the first through hole and the second through hole are aligned one by one.
[0009] This invention also provides a method for batch staining of ultrathin electron microscopy sections, characterized in that... Batch staining specifically includes the following steps: Step 1: First, pull the bottom box out of the lid. Then, vertically place the multiple screens containing the ultrathin sections to be stained into the placement slots of the bottom box, so that the screens are positioned at two opposite corners of the placement slots. Then, insert the bottom box back into the lid. Step 2: Place the copper mesh box into the groove of the wax tray and add a certain amount of uranium acetate staining solution into the groove of the wax tray. The uranium acetate staining solution penetrates evenly into the surface of each ultrathin section of the carrier mesh through the aligned small holes. After covering the tray, let it stand for a period of time for staining and perform the first staining. Step 3: After the first dyeing is completed, the copper mesh box is moved to a beaker containing distilled water for washing using the handle. During the washing process, the operator operates the handle to shake the copper mesh box up and down in the distilled water a certain number of times to wash away excess uranium acetate. During the up and down shaking, the mesh is kept stuck in the placement tank, and the wax tray is cleaned at the same time. Step 4: Place the washed copper mesh box back into the groove of the wax tray, add a certain amount of lead citrate dye solution into the groove of the wax tray, and place an appropriate amount of sodium hydroxide solid around the wax tray. After covering the tray, let it stand for a period of time to perform the second dyeing.
[0010] Step 5: After the second dyeing is completed, move the copper mesh box to a beaker containing fresh distilled water for washing. During the washing process, the operator operates the handle to shake the copper mesh box up and down in the distilled water a certain number of times to wash away excess lead citrate. During the up and down shaking, the mesh is firmly stuck in the placement tank, and the wax tray is cleaned. Step 6: After washing, remove the bottom box from the lid, use tweezers to remove the carrier net from the bottom box and dry it to complete the dyeing process.
[0011] The advantages of this invention application compared to the prior art are: 1) This invention uses a manual staining method, which is small in size, simple to operate, and particularly suitable for laboratories in schools or research institutions, and has low cost; 2) The copper mesh box of the present invention adopts a bottom box that is snapped onto the lid. Multiple placement slots are provided on the bottom box, and the placement slots are diamond-shaped, which can vertically hold the mesh in the two opposite corners of the placement slots. This allows both sides of the mesh to come into contact with the dye solution, and the vertical placement makes it easy to remove the mesh from the placement slots. Perforations are provided on the upper and lower sides of the placement slots. The mesh is held between the two perforations in the vertical direction. This ensures that the mesh is cleaned while also fixing it during washing, preventing the mesh from moving back and forth or even falling out of the placement slots during washing. Attached Figure Description
[0012] Figure 1 This is a three-dimensional cross-sectional view of a batch staining device for ultrathin electron microscopy section samples according to the present invention.
[0013] Figure 2 yes Figure 1 A magnified view of a portion of point A in the middle.
[0014] Figure 3 This is a three-dimensional view of the copper mesh box and wax tray in a batch staining device for ultrathin section samples for electron microscopy, as described in this invention application.
[0015] Figure 4 This is a three-dimensional view of the copper mesh box in a batch staining device for ultrathin section samples for electron microscopy, as described in this invention application.
[0016] Figure 5 yes Figure 4 A magnified view of a portion of point B in the middle.
[0017] Figure 6This is a planar cross-sectional view of the copper mesh box and wax tray in a batch staining device for ultrathin section samples for electron microscopy, as described in this invention application.
[0018] Figure 7 This is a three-dimensional view of the wax tray in a batch staining device for ultrathin section samples for electron microscopy, as described in this invention application.
[0019] Figure 8 This is a perspective view of the tray, tray cover, copper mesh box, handle, and wax tray in a batch staining device for ultrathin section samples for electron microscopy according to the present invention. Detailed Implementation
[0020] To further understand the features, technical means, and specific objectives and functions achieved by this invention application, the invention application will be described in further detail below with reference to the accompanying drawings and specific embodiments.
[0021] Reference Figures 1 to 8 The image shows a batch staining device for ultrathin sections of electron microscope samples, comprising a tray 1, a tray cover 2, a copper mesh box 3, a handle 4, and a wax tray 5. The copper mesh box 3 includes a base box 31 and a cover 32. The base box 31 has multiple placement slots 311, which are rhomboid in top view. The placement slots 311 are used to place the mesh carrier 6, which can be placed vertically within the placement slots 311, and the mesh carrier 6 is precisely positioned within the two opposite angles of the rhombus shape of the placement slot 311. A first perforation 312 is provided at the bottom of each placement slot 311, penetrating the base box 31 vertically. The diameter of the first perforation 312 is smaller than the diameter of the mesh carrier 6. A second perforation 322 is provided on the cover 32 at the position corresponding to each placement slot 311, and the diameter of the second perforation 322 is smaller than the diameter of the mesh carrier 6. The handle 4 is fixedly connected to the cover 32 of the copper mesh box 3, facilitating operation by the operator. The wax tray 5 is placed on the tray 1, and the wax tray 5 has a groove 51 that matches the shape of the copper mesh box 3, allowing the copper mesh box 3 to be placed within the groove 51. The tray cover 2 is used to cover the tray 1 during dyeing to reduce the influence of light and carbon dioxide in the air on the dyeing process.
[0022] Furthermore, the bottom box 31 is snapped onto the lid 32. Specifically, a retaining strip 323 is provided on two opposite sides of the lid 32 and located below the lid 32, forming a retaining groove 324 between the retaining strip 323 and the lid 32. An insertion part 313 is provided on both sides of the corresponding groove 324 of the bottom box 31, and the insertion part 313 can be inserted into the groove 324 to achieve the snapping of the bottom box 31 and the lid 32.
[0023] An abutment strip 314 extending along the insertion direction is provided on the lower surface of one of the insertion parts 313. The abutment strip 314 can abut against the corresponding card strip 323. On the one hand, it can reduce the contact area between the insertion part 313 and the card strip 323, making it easier for the insertion part 313 to be inserted into the card slot 324. On the other hand, the abutment strip 314 and the card strip 323 abut against each other to prevent the bottom box 31 from falling off the box cover 32 on its own.
[0024] Batch staining specifically includes the following steps: Step 1: First, pull the bottom box 31 out from the lid 32, and vertically place the multiple screens 6 containing the ultrathin slices to be stained into the placement slots 311 of the bottom box 31, so that the screens 6 are stuck at the two opposite corners of the placement slots 311; then insert the bottom box 31 into the lid 32. Step 2: Place the copper mesh box 3 into the groove 51 of the wax tray 5, and add a certain amount of uranium acetate staining solution into the groove 51 of the wax tray 5. The uranium acetate staining solution penetrates evenly into the surface of the ultrathin section of each carrier mesh 6 through the vertically aligned small holes 312 and 322. After covering the tray cover 2, let it stand for a period of time for staining and perform the first staining. Step 3: After the first dyeing is completed, the copper mesh box 3 is moved to a beaker containing distilled water by the handle 4 for washing. During the washing, the operator operates the handle 4 to shake the copper mesh box 3 up and down in the distilled water a certain number of times to wash away excess uranium acetate. During the up and down shaking, the mesh is stuck in the placement tank 311, and the wax plate 5 is cleaned at the same time. Step 4: Place the washed copper mesh box 3 back into the groove 51 of the wax tray 5, add a certain amount of lead citrate dye solution into the groove 51 of the wax tray 5, and place an appropriate amount of sodium hydroxide solid around the wax tray 5. After covering with the tray cover 2, let it stand for a period of time to perform the second dyeing.
[0025] Step 5: After the second dyeing is completed, move the copper mesh box 3 to a beaker containing fresh distilled water for washing. During the washing, the operator operates the handle 4 to shake the copper mesh box 3 up and down in the distilled water a certain number of times to wash away excess lead citrate. During the up and down shaking, the mesh is stuck in the placement tank 311, and the wax tray 5 is cleaned. Step 6: After washing, remove the bottom box 31 from the box cover 32, and use tweezers to remove the carrier net 6 from the bottom box 31 for drying to complete the dyeing process.
[0026] The above embodiments only illustrate one or more implementation methods of this invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of this invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this invention, and these all fall within the protection scope of this invention. Therefore, the protection scope of this invention should be determined by the appended claims.
Claims
1. A batch staining device for ultrathin electron microscopy sections, characterized in that, include: Tray (1); A wax plate (5) is placed on the tray (1), the wax plate (5) is open to the upward, and a groove (51) is formed on the wax plate (5). A copper mesh box (3) is used to hold multiple carrier meshes (6), each carrier mesh (6) having an ultrathin slice to be stained placed on it, and the copper mesh box (3) can be placed into the groove (51); A handle (4) is provided on the copper mesh box (3) for taking the copper mesh box (3) out and putting it in; The tray cover (2) is opaque and is used to cover the tray (1).
2. The batch staining apparatus for ultrathin electron microscopy sections according to claim 1, characterized in that, The copper mesh box (3) includes a bottom box (31) and a lid (32). The bottom box (31) has multiple placement slots (311). When viewed from above, the placement slots (311) are rhomboid. The placement slots (311) are used to place the carrier net (6). The carrier net (6) can be placed vertically in the placement slots (311), and the carrier net (6) is just stuck in the two opposite angles of the rhomboid shape of the placement slots (311). The handle (4) is fixedly connected to the lid (32) of the copper mesh box (3).
3. The batch staining device for ultrathin electron microscopy sections according to claim 2, characterized in that, A first through hole (312) is provided at the bottom of each placement slot (311). The first through hole (312) penetrates the bottom box (31) vertically. The diameter of the first through hole (312) is smaller than the diameter of the carrier net (6). A second perforation (322) is provided on the cover (32) and at the position corresponding to each placement slot (311), the diameter of the second perforation (322) being smaller than the diameter of the carrier net (6).
4. The batch staining device for ultrathin electron microscopy sections according to claim 3, characterized in that, The bottom box (31) is snapped onto the lid (32).
5. The batch staining apparatus for ultrathin electron microscopy sections according to claim 4, characterized in that, On the two opposite sides of the lid (32) and below the lid (32), there are locking strips (323) and locking grooves (324) formed between the locking strips (323) and the lid (32); on both sides of the corresponding locking grooves (324) of the bottom box (31), there are insertion parts (313) that can be inserted into the locking grooves (324) to achieve the locking of the bottom box (31) and the lid (32). When the bottom box (31) is inserted into the lid (32), the placement groove (311), the first through hole (312) and the second through hole (322) are aligned one by one.
6. A method for batch staining of ultrathin electron microscopy sections, using the batch staining apparatus for ultrathin electron microscopy sections according to claim 5, characterized in that, Specifically, the steps include the following: Step 1: First, pull the bottom box out of the lid. Then, vertically place the multiple screens containing the ultrathin sections to be stained into the placement slots of the bottom box, so that the screens are positioned at two opposite corners of the placement slots. Then, insert the bottom box back into the lid. Step 2: Place the copper mesh box into the groove of the wax tray and add a certain amount of uranium acetate staining solution into the groove of the wax tray. The uranium acetate staining solution penetrates evenly into the surface of each ultrathin section of the carrier mesh through the aligned small holes. After covering the tray, let it stand for a period of time for staining and perform the first staining. Step 3: After the first dyeing is completed, the copper mesh box is moved to a beaker containing distilled water for washing using the handle. During the washing process, the operator operates the handle to shake the copper mesh box up and down in the distilled water a certain number of times to wash away excess uranium acetate. During the up and down shaking, the mesh is kept stuck in the placement tank, and the wax tray is cleaned at the same time. Step 4: Place the washed copper mesh box back into the groove of the wax tray, add a certain amount of lead citrate dye solution to the groove of the wax tray, and place an appropriate amount of sodium hydroxide solid around the wax tray. After covering the tray, let it stand for a period of time for dyeing, and then perform the second dyeing. Step 5: After the second dyeing is completed, move the copper mesh box to a beaker containing fresh distilled water for washing. During the washing process, the operator operates the handle to shake the copper mesh box up and down in the distilled water a certain number of times to wash away excess lead citrate. During the up and down shaking, the mesh is firmly stuck in the placement tank, and the wax tray is cleaned. Step 6: After washing, remove the bottom box from the lid, use tweezers to remove the carrier net from the bottom box and dry it to complete the dyeing process.