A hard tissue section anti - detachment pretreatment device and an anti - detachment pretreatment method

Abstract

The present invention discloses a pretreatment device for preventing hard tissue sections from falling off, which includes a box body and a section rack. A buffer plate is provided inside the box body, and a plurality of small holes are evenly arranged on the buffer plate; the bottom of the box body is communicated with a drain pipe through a liquid outlet, and a valve is provided on the drain pipe; the section rack is placed on the buffer plate, and the section rack includes two longitudinal support frames and multiple layers of transverse support frames fixed between the two longitudinal support frames; each layer of transverse support frame includes multiple first support rods, and the first support rods have a slope of 0 to 15° relative to the horizontal plane. One end of the first support rod with a lower horizontal height extends upward to form a baffle, and the other end of the baffle is fixed to a second support rod. The present invention provides a specific device for application to the process of removing the plastic film before staining hard tissue sections, reducing the probability of staining and section detachment from 70% to less than 10%, greatly improving the staining success rate of hard tissue sections, and providing a huge room for improvement for the staining of hard tissue sections.

Description

Technical Field

[0001] The present invention belongs to the technical field of slice anti - detachment, and particularly relates to a hard tissue slice anti - detachment pretreatment device and an anti - detachment pretreatment method. Background Art

[0002] The existing anti - detachment glass slides on the market, including positively charged anti - detachment glass slides, lysine - treated anti - detachment glass slides, etc., all increase the anti - detachment performance of the glass slides by performing surface anti - detachment treatment on the glass slides. These anti - detachment glass slides have good anti - detachment effects on paraffin sections and frozen sections. However, even the very expensive anti - detachment glass slides have not been able to solve the problem of section detachment for hard tissue sections well. These anti - detachment glass slides are expensive, but have no obvious anti - detachment effect on hard tissue sections, and there is still a detachment rate of more than 60% - 70%.

[0003] The high detachment rate of hard tissue section staining greatly reduces the staining success rate of hard tissue sections, resulting in an obstacle to the development of hard tissue section staining, and has always been the main obstacle to the development and progress of hard tissue sections.

[0004] Therefore, providing a technical means to reduce the detachment rate of hard tissue section staining and improve the staining success rate of hard tissue sections is an important research direction. Summary of the Invention

[0005] Aiming at the above - mentioned defects of the prior art, a hard tissue slice anti - detachment pretreatment device and an anti - detachment pretreatment method are provided.

[0006] In the first aspect of the present invention, a hard tissue slice anti - detachment pretreatment device is provided, which includes a box body and a slice rack. A buffer plate is arranged in the box body, and a plurality of small holes are evenly arranged on the buffer plate; a box cover is arranged above the box body, and the bottom of the box body is communicated with a drain pipe through a liquid outlet, and a valve is arranged on the drain pipe;

[0007] The slice rack is placed on the buffer plate. The slice rack includes two longitudinal support frames and multiple layers of transverse support frames fixed between the two longitudinal support frames; each layer of the transverse support frame includes multiple first support rods, and the first support rods have a slope of 0 - 15° relative to the horizontal plane. One end of the first support rod with a lower horizontal height extends upward to form a baffle, and the other end of the baffle is fixed to a second support rod;

[0008] Vertical support rods are arranged at both ends of the longitudinal support frame, and multiple second support rods are fixedly arranged in parallel between the two vertical support rods. The same ends of the first support rods in the same layer are fixed to the same second support rod.

[0009] Preferably, the number of layers of the transverse support frame is 3 - 8 layers.

[0010] Preferably, a liquid discharge channel extends downward from the bottom of the box body from the liquid outlet, and the lower end of the liquid discharge channel communicates with the drain pipe.

[0011] Preferably, the diameter of the lower end of the liquid discharge channel gradually decreases to be the same as that of the drain pipe.

[0012] Preferably, the box body and the slicing rack are made of glass, stainless steel or polytetrafluoroethylene.

[0013] Preferably, a valve seat is provided on the drain pipe, the valve is sleeved in the valve seat, a through hole is provided on the valve, and the valve is rotated in the valve seat, and the through hole communicates with or disconnects from the drain pipe.

[0014] Preferably, the diameter of the small holes on the buffer plate is 0.5 mm to 3 mm.

[0015] In a second aspect, the present invention provides a method for preventing tissue sections from falling off during pretreatment, using the above-mentioned device for preventing tissue sections from falling off during pretreatment, which specifically includes the following steps:

[0016] Step S1: Place the hard tissue section on the slicing rack and place the slicing rack in the box body;

[0017] Step S2: After confirming that the valve is closed, pour the first stripping solution from above the box body until the hard tissue section is immersed in the first stripping solution;

[0018] Step S3: After soaking for a period of time, open the valve to collect the first stripping solution;

[0019] Step S4: Pour the second stripping solution, the anhydrous ethanol-acetone mixture and anhydrous ethanol in sequence, and repeat the operations in Steps S2-S3;

[0020] Step S5: Take out the hard tissue section, seal the section with dropping collodion to obtain a hard tissue section subjected to anti-falling pretreatment.

[0021] The present invention adopts the above technical solutions, compared with the prior art, has the following technical effects:

[0022] (1) By providing a specific device for use in the stripping process before staining of hard tissue sections, the probability of staining and section detachment of the present invention is reduced from 70% to less than 10%, greatly improving the staining success rate of hard tissue sections and providing a huge improvement space for the staining of hard tissue sections.

[0023] (2) In the present invention, a buffer plate with dense holes is arranged inside the box body to change the flow direction of the stripping solution, the mixed solution of anhydrous ethanol and acetone, anhydrous ethanol, etc. When flowing downward through the guidance of the dense small holes, the liquid in the box body will only generate small vortices with relatively light fluctuations and will not cause large fluid fluctuations in the box body to cause film detachment.

[0024] (3) This device can be applied not only to the pre-treatment process before staining of hard tissue sections, but also to the pre-treatment process before staining of paraffin sections and frozen sections.

[0025] (4) The anti-detachment method provided by the present invention is different from the existing technical methods of adding anti-detachment reagents or performing anti-detachment treatment on the surface of the glass slide, with low cost and simple operation, achieving a very good anti-detachment effect. Brief Description of the Drawings

[0026] Figure 1 It is the front view of the section rack in the present invention;

[0027] Figure 2 It is the top view of the section rack in the present invention;

[0028] Figure 3 It is Figure 1 the schematic structural diagram of the A-A cross-section in

[0029] Figure 4 It is the three-dimensional view of the section rack in the present invention;

[0030] Figure 5 It is the schematic structural diagram of a hard tissue section anti-detachment pre-treatment device in the present invention;

[0031] Figure 6 It is the front view of a hard tissue section anti-detachment pre-treatment device in the present invention;

[0032] Figure 7 It is Figure 6 the schematic structural diagram of the B-B cross-section in

[0033] Figure 8 It is the schematic structural diagram of the valve in the present invention;

[0034] The notations in the figures indicate:

[0035] 1 - box body, 11 - buffer plate, 12 - box cover, 13 - liquid outlet, 2 - section rack, 21 - longitudinal support frame, 211 - vertical support rod, 212 - second support rod, 22 - transverse support frame, 221 - first support rod, 3 - drain pipe, 4 - valve. Detailed Embodiments

[0036] When making slices or ultra-thin slices, it is difficult to make slices of uniform thickness because the tissue is soft or the hardness is uneven locally. Therefore, it is necessary to use a certain substance to penetrate the inside of the tissue to make the entire tissue harden uniformly to facilitate cutting into thin slices. This substance is called an embedding agent. Generally, slices observed under an optical microscope use paraffin, collodion, charcoal wax, gelatin, etc. as embedding agents; electron microscopes use epoxy resins, polystyrene resins, isobutylene resins and water-soluble resins.

[0037] Before HE staining, special staining or immunohistochemical staining, the slices must be de-plasticized. The de-plasticization rate of hard tissue slices after de-plasticization by traditional de-plasticization methods is very high during the staining process, reaching about 70%. The present invention adopts a hard tissue slice de-plasticization pretreatment device, such as Figures 5 - 7 As shown in , the pretreatment process before dyeing is improved to achieve the technical purpose of preventing shedding.

[0038] The present invention is described in detail and specifically by specific examples below to provide a better understanding of the present invention, but the following examples do not limit the scope of the present invention.

[0039] Example 1

[0040] See also Figures 5 - 7 As shown in , this embodiment provides a hard tissue slice anti-falling pretreatment device, including a box body 1 and a slice rack 2.

[0041] like Figure 7 In the embodiment, a buffer plate 11 is provided in the box body 1 for carrying the slice rack 2. In a specific embodiment, the buffer plate 11 is a plate-like structure with a plurality of holes, which is horizontally fixed at the lower part of the box body 1. The buffer plate 11 divides the box body 1 into an upper space and a lower space. The liquid flowing from the top of the box body 1 flows from the upper space to the lower space through the holes on the buffer plate 11. Preferably, the diameter of the small holes on the buffer plate (11) is 0.5 mm to 3 mm. In the present invention, by providing a buffer plate 11 with dense holes in the box body 1, the flow direction of the plastic removal liquid, the anhydrous ethanol acetone mixed liquid, the anhydrous ethanol, etc. when discharged is changed; through the guidance of the dense small holes, the liquid in the box body 1 will only generate a small vortex with relatively light fluctuation when flowing downward, and will not generate a large fluid fluctuation in the box body to cause the slice to be detached. If the buffer plate 11 is not provided, and the liquid is only discharged through the discharge port of the box bottom 1, a large fluid spiral will be generated, causing a relatively strong fluid fluctuation in the box body, resulting in the slice to be detached in the subsequent operation.

[0042] In this embodiment, if Figure 7 As shown in FIG. 1 , the buffer plate 11 is horizontally fixed at a position 10 mm away from the bottom of the box body, and the thickness of the buffer plate 11 is 2.5 mm. This size is only a specific embodiment of the present invention, and the present invention is not limited to the above size.

[0043] In a preferred embodiment, the box body 1 and the buffer plate 11 are integrally provided.

[0044] In this embodiment, as Figure 6 shown, a box cover 12 is provided above the box body 1, and the bottom of the box body 1 is communicated with a drain pipe 3 through a liquid outlet 13. A valve 4 is provided on the drain pipe 3; specifically, as Figure 6 shown, a valve seat is provided on the drain pipe 3, and the valve 4 is sleeved in the valve seat. As Figure 8 shown, a through hole is provided on the valve 4. By rotating the knob on the valve 4, the valve 4 rotates in the valve seat. When it is necessary to drain the liquid in the box body 1, rotate the valve 4 until the through hole communicates with the drain pipe. The flow rate of the drained liquid can also be adjusted by rotating the knob on the valve 4 to further control the fluid fluctuation in the box body 1.

[0045] Specifically, as Figures 6 - 7 shown, the length of the box body is 100 mm, the width is 70 mm, the internal width is 66 mm, the wall thickness is 2 mm, and the height is 70 mm; the above dimensions are only a specific embodiment of the present invention, and the present invention is not limited to the above dimensions.

[0046] In this embodiment, as Figure 7 shown, the pipe diameter of the drain pipe 3 is 7 mm. The above dimensions are only a specific embodiment of the present invention, and the present invention is not limited to the above dimensions.

[0047] In this embodiment, as Figure 4 shown, the slicing rack 2 includes two longitudinal support frames 21 and multiple layers of transverse support frames 22 fixed between the two longitudinal support frames 21. As Figure 2 shown, each layer of the transverse support frame 22 includes multiple parallel first support rods 221; specifically, as Figure 2 shown, the transverse support frame 22 has four first support rods 221. The distance between adjacent first support rods 221 is 18 mm, the maximum distance between the first support rods 221 is 48 mm, and the length of the transverse support frame 22 is 77 mm; as Figure 3 shown, the distance between adjacent layers of the transverse support frames 22 is 7 mm, the height of the first support rods 221 is 3 mm, and the length of the straight section of the first support rods 221 is 75 mm; the above dimensions are only a specific embodiment of the present invention, and the present invention is not limited to the above dimensions.

[0048] In this embodiment, as Figure 4 shown, vertical support rods 211 are provided at both ends of the longitudinal support frame 21. Multiple second support rods 212 are fixedly arranged in parallel between the two vertical support rods 211. The same ends of the first support rods 221 in the same layer are fixed on the same second support rod 212. As Figure 3Among them, the vertical support rod 211 has a height of 50 mm, a width of 3 mm, and the total width of the slicing rack 2 is 80 mm; the above dimensions are only a specific embodiment of the present invention, and the present invention is not limited to the above dimensions.

[0049] In this embodiment, as Figure 3 Among them, one end of the first support rod 221 is fixed on the side of the second support rod 212 facing the first support rod 221, and the other end extends upward to form a baffle and is fixed on the lower surface of the second support rod 212. The first support rod 221 has a slope of 0 to 15° relative to the horizontal direction, and the end with the baffle is lower than the other end, that is, the heights of the second support rods 212 at both ends of the same first support rod 221 can be the same or different. In a preferred embodiment, the height of the baffle on the lower side is 1 mm to 3 mm to ensure that the slices will not easily slide out of the track. The baffle can be arc-shaped or perpendicular to the first support rod 221.

[0050] In this embodiment, as Figure 2 Among them, the number of layers of the horizontal support frame 22 is 3 to 8 layers, and further preferably, it is 5 layers, and the height between layers is 7 mm. In specific applications, it can be freely selected according to the number of glass slides.

[0051] In this embodiment, as Figure 5 Among them, in a preferred embodiment, the bottom of the box body 1 extends downward from the liquid outlet 13 to form a liquid outlet channel, and the diameter of the lower end of the liquid outlet channel gradually decreases to be the same as that of the drain pipe 3, and the lower end of the liquid outlet channel is communicated with the drain pipe 3.

[0052] In a specific embodiment, the materials of the box body 1 and the slicing rack 2 are glass, stainless steel or polytetrafluoroethylene; further, for cost considerations, the material is glass.

[0053] In a preferred embodiment, the length, width and height of the box body 1, the length, width and height of the slicing rack 2, and the number of layers can be selected according to the number of slices to be processed.

[0054] Embodiment 2

[0055] The present invention provides a method for preventing hard tissue slices from falling off during pretreatment. Using the hard tissue slice anti-detachment pretreatment device provided in Embodiment 1, taking the pretreatment before HE staining of hard tissue slices as an example, it specifically includes the following steps:

[0056] Step S1: Place the hard tissue slices on the slicing rack 2, 2 slices per layer, a total of 10 slices in five layers (which can be adjusted according to the actual number of slices), and place the slicing rack 2 in the box body 1;

[0057] Step S2: After confirming that the valve 4 is closed, pour the first stripping solution from the upper part of the box body 1 until the hard tissue slices are immersed in the first stripping solution;

[0058] Step S3: After 10 minutes of plastic stripping, open valve 4 to allow the first plastic stripping liquid to slowly flow into the collection bottle. After all the first plastic stripping liquid has flowed out, tighten valve 4 again.

[0059] Step S4: Pour in the second plastic stripping liquid, a 1:1 mixture of anhydrous ethanol and acetone, and anhydrous ethanol in sequence, and repeat the operations in Steps S2 - S3.

[0060] Step S41: Pour in the second plastic stripping liquid and perform plastic stripping for 10 minutes. Open valve 4 to allow the second plastic stripping liquid to slowly flow into the collection bottle. After all the second plastic stripping liquid has flowed out, tighten valve 4 again.

[0061] Step S42: Pour in the 1:1 mixture of anhydrous ethanol and acetone and soak for 2 minutes. Open valve 4 to allow the 1:1 mixture of anhydrous ethanol and acetone to slowly flow into the collection bottle. After all the 1:1 mixture of anhydrous ethanol and acetone has flowed out, tighten valve 4 again.

[0062] Step S43: Pour in anhydrous ethanol and soak for 2 minutes. Open valve 4 to allow anhydrous ethanol to slowly flow into the collection bottle. After all the anhydrous ethanol has flowed out, tighten valve 4 again.

[0063] Step S5: Take out the hard tissue section, and drop collodion with a concentration of 0.25% - 5% onto the observation area of the section for sealing to obtain a hard tissue section that has undergone anti - detachment pretreatment.

[0064] The specific embodiments of the present invention have been described in detail above, but they are only examples, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions made to the present invention are also within the scope of the present invention. Therefore, all equivalent transformations and modifications made without departing from the spirit and scope of the present invention should be covered within the scope of the present invention.

Claims

1. An anti - detachment pretreatment device for the process of removing plastic before hard tissue section staining, characterized in that, It includes a box body (1) and a slicing rack (2). A buffer plate (11) is provided inside the box body (1), and a plurality of small holes are evenly arranged on the buffer plate (11); a box cover (12) is provided above the box body (1), and the bottom of the box body (1) is communicated with a drain pipe (3) through a liquid outlet (13), and a valve (4) is provided on the drain pipe (3); The slicing rack (2) is placed on the buffer plate (11). The slicing rack (2) includes two longitudinal support frames (21) and multiple layers of transverse support frames (22) fixed between the two longitudinal support frames (21); each layer of the transverse support frame (22) includes multiple first support rods (221), and the first support rods (221) have a slope of 0 to 15° relative to the horizontal plane. One end with a lower horizontal height of the first support rod (221) extends upward to form a baffle, and the other end of the baffle is fixed to a second support rod (212); Vertical support rods (211) are provided at both ends of the longitudinal support frame (21), and multiple second support rods (212) are fixedly arranged in parallel between the two vertical support rods (211). The same end of the first support rods (221) in the same layer is fixed to the same second support rod (212); The baffle is arc-shaped.

2. The anti - detachment pretreatment device for the demolding process before hard tissue section staining according to claim 1, characterized in that, The height of the arc-shaped baffle is 1 mm to 3 mm.

3. The anti - detachment pretreatment device for the decalibration process before hard tissue section staining according to claim 1, wherein, The buffer plate (11) is horizontally fixed at a position 8 to 15 mm away from the bottom of the box body (1).

4. The anti - detachment pretreatment device for the decalcification process before hard tissue section staining according to claim 1, characterized in that, The number of layers of the transverse support frame (22) is 3 to 8 layers.

5. The anti - detachment pretreatment device for the decalcification process before hard tissue section staining according to claim 1, characterized in that, The bottom of the box body (1) extends downward from the liquid outlet (13) to form a liquid outlet channel, and the lower end of the liquid outlet channel is communicated with the drain pipe (3).

6. The anti - detachment pretreatment device for the decalibration process before hard tissue section staining according to claim 5, characterized in that, The diameter of the lower end of the liquid outlet channel gradually decreases to be the same as that of the drain pipe (3).

7. The anti - detachment pretreatment device for the decalcification process before hard tissue section staining according to claim 1, wherein, The materials of the box body (1) and the slicing rack (2) are glass, stainless steel or polytetrafluoroethylene.

8. The anti - detachment pretreatment device for the plastic - removing process before hard tissue section staining according to claim 1, characterized in that, A valve seat is provided on the drain pipe (3), the valve (4) is sleeved in the valve seat, and a through hole is provided on the valve (4). By rotating the valve (4), the through hole is communicated with or disconnected from the drain pipe (3).

9. The anti - detachment pretreatment device for the plastic - removing process before hard tissue section staining according to claim 1, characterized in that, The diameter of the small holes on the buffer plate (11) is 0.01 mm to 3 mm.

10. A method for preprocessing to prevent hard tissue sections from falling off, using the anti-falling-off preprocessing device according to any one of claims 1-9, characterized in that, Specifically, it includes the following steps: Step S1: Place the hard tissue slice on the slicing rack (2), and place the slicing rack (2) in the box body (1); Step S2: After confirming that the valve (4) is closed, pour the first deplasticizing liquid from above the box body (1) until the hard tissue slice is immersed in the first deplasticizing liquid; Step S3: After soaking for a period of time, open the valve (4), collect the liquid after the first deplasticization, and then close the valve (4); Step S4: Pour the second deplasticizing liquid, anhydrous ethanol-acetone mixture and anhydrous ethanol in sequence, and repeat the operations in steps S2 and S3; Step S5: Take out the hard tissue slice, and perform sealing with dropping collodion to obtain a hard tissue slice pretreated against detachment.

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