CN104698163B - The using method of the detachable also microslide couveuse of volume adjustable - Google Patents

Abstract

The invention discloses a detachable and volume-adjustable glass slide incubator. The stainless steel incubation frame is pressed on the glass slide, and together with the silica gel cushion layer below it, an incubation chamber is formed from a rectangular space in the middle with the glass slide as the bottom surface; In addition, there is an elastic holder for clamping the stainless steel incubation frame and the silicone cushion layer on the glass slide; the inner walls of the stainless steel incubation frame and the rubber cushion layer are respectively provided with a dovetail groove, and the dovetail groove is connected with a dovetail-shaped convex body. The incubator space adjustment block plug fits. The invention has a reasonable structure, which ensures that when the antibody is incubated, the effect that all tissue sections pasted on the glass slide can be completely soaked in the antibody incubation solution is obtained by using a smaller amount of antibody incubation solution, further saving the cost of scientific research. Experimental cost.

Description

How to use the detachable and volume-adjustable slide incubator

This application is a divisional application with application number: 201410012882.3, application date: 2014.1.13, and name "Detachable and Volume-Adjustable Slide Incubator".

technical field

The invention relates to a detachable slide incubator with adjustable volume.

Background technique

In life science research, people often need to use immunohistochemical techniques with strong specificity and high sensitivity to locate and quantify specific substances in tissue cells. The basic principle of immunohistochemical technology is to first use known immunological antibodies (primary antibodies) to bind to the antigen to be recognized in the observed tissue. Since the antigen-antibody binding reaction on tissue cells is generally invisible, it is necessary to use a labeling method to link a certain marker (such as an enzyme, fluorescein) to the primary antibody, and then use histochemical methods to display this. label or observe fluorescence from fluorescein under fluorescence microscopy, confocal laser microscopy, or super-resolution microscopy. To visualize certain substances in the cells of the observed tissue. When implementing the above-mentioned labeling method to bind a certain marker (such as an enzyme, fluorescein) to the first antibody, people often use the specificity of the second antibody that has been bound to a certain marker and the first antibody. method of sexual union.

When immunohistochemical techniques are used to locate and quantify specific substances in tissue cells, patch staining is usually used to maintain the good shape of experimental tissue sections and to clarify the order of each tissue section. The basic principle of the patch staining method is to paste the tissue slices on the glass slide in the order of sectioning immediately after cutting the tissue slices on the microtome. Subsequent blocking of the tissue section, incubation with the first antibody, washing, incubation with the second antibody, re-washing, dehydration, transparency and sealing are all carried out under the condition that the tissue section is pasted on the glass slide.

In the process of sealing, washing, rewashing, dehydration, transparent and sealing of the above-mentioned tissue sections, the cost of reagents for preparing these solutions is relatively low, and in addition, the amount of washing solution is relatively large during the washing process. In order to obtain a good experimental effect, people usually place the entire glass slide pasted with tissue sections in the above-mentioned relevant solution for processing. However, when incubating with the first antibody and the second antibody, due to the high price of the antibody (currently, the price of 100 μl of a general monoclonal primary antibody is 3,000 yuan, and the price of a special monoclonal primary antibody will be higher), and The multiples that can be diluted during use cannot be too large. Therefore, antibodies, especially primary antibodies, become a major part of the cost of immunohistochemistry experiments. Therefore, when immunohistochemical techniques are used to locate and quantify specific substances in tissue cells, it cannot be done by placing the entire glass slide pasted with tissue sections in the antibody incubation solution.

When people perform immunohistochemical staining in the patch method, the method of directly adding the antibody incubation solution to the tissue section of the glass slide is usually used. However, when using this method, if the antibody incubation solution is added too much, it will not be preserved and will spill out of the glass slide, and if the antibody incubation solution on the tissue section is added less, it will not be preserved due to the evaporation of water in the incubation solution during the incubation process. Tissue sections on glass slides are always soaked in antibody incubation solution. Sometimes, people also use the method of drawing a frame with oily material around the tissue section attached to the glass slide, and add antibody incubation solution in the oily frame to prevent the antibody incubation solution from overflowing. However, although this method has some improvements over the method of directly adding the antibody incubation solution to the slide-attached tissue section, using this method, due to the poor ability of the oily frame to prevent the antibody incubation solution from spilling, it cannot guarantee During the entire antibody incubation process, the tissue sections were kept soaked in the antibody incubation solution. Moreover, in the process of antibody incubation, washing, dehydration, and transparency, these oily materials will lose the effect of preventing the antibody incubation solution from overflowing during antibody incubation due to falling off; when oily materials fall off and adhere to tissue sections, it will affect the final results. Moreover, during the processing of tissue slices, oily materials will also dissolve into related experimental solutions, polluting the experimental solutions and affecting the reuse of experimental solutions and the processing effect of tissue slices.

Contents of the invention

The object of the present invention is to provide a detachable and volume-adjustable slide incubator with reasonable structure, convenient and efficient implementation of all experimental steps, and good immunohistochemical staining experimental results.

Technical solution of the present invention is:

A detachable and volume-adjustable slide incubator, which is characterized by: including a stainless steel incubation frame, which is pressed on the slide glass, and together with the silica gel cushion layer below, the rectangular space in the middle is formed by the slide glass. The bottom surface forms an incubation chamber; the silica gel pad has the same middle rectangular space as the stainless steel incubation frame, and adheres to the stainless steel incubation frame to contact with the glass slide to prevent the leakage of the incubation solution; the outer dimension of the stainless steel incubation frame and the silica gel The peripheral dimensions of the cushion layer are all the same as the peripheral dimensions of the slide glass; an elastic holder is provided to clamp the stainless steel incubation frame and the silica gel cushion layer on the glass slide; the elastic holder includes the root of the elastic holder, The clamping part of the elastic holder and the clamping port of the elastic holder; the root of the elastic holder is connected to the clamping part of the elastic holder, and the elastic holder is provided for forming the slide incubator and removing the slide The hand-held position when the incubator is in operation; the elastic holder is clamped on the incubation frame and the glass slide through the clamping port of the elastic clamp, and the clamping part of the elastic clamp clamps the incubation frame and the glass slide on the Together to form an incubation chamber; the stainless steel incubation frame and the inner wall of the silicone cushion are respectively provided with a dovetail groove on the four sides, and the dovetail groove is plugged with the incubation chamber space adjustment block with a dovetail-shaped convex body to adjust the size of the incubation chamber and prevent The incubation chamber space adjustment block moves in the incubation chamber.

The method of use is: Laminate the stainless steel incubation frame and silica gel pad on the glass slide with tissue slices attached to ensure that all tissue slices are at the bottom of the incubation chamber formed by the incubation frame and slide glass; select the incubation frame and slide glass Overlap any one of the four sides up and down, hold the root of the elastic holder, align the clamping mouth of the elastic holder with the structure where the incubation frame and the slide slide are stacked up and down, push the elastic holder into the incubation frame and A structure in which glass slides are stacked on top of each other. Ensure that the clamping part of the elastic holder tightly presses the incubation frame and the glass slide together, and repeat the above-mentioned action on the other three sides to clamp the elastic holder in to ensure that the incubation frame is in close contact with the slide to form an inseparable Leaky incubation chambers; after forming a non-leaking incubation chamber, selectively insert 1 to 4 incubation chamber spaces into the 4 dovetail grooves on the rectangular wall of the incubation frame, depending on where the tissue section is attached on the slide Adjustment block to flexibly adjust the size of any side or polygonal space in the incubation frame; after adjusting the size of the incubation chamber space, add the incubation solution of the primary antibody to the incubation chamber to ensure that the tissue slices pasted on the bottom of the incubation chamber can be soaked In the incubation solution containing the specific first antibody; incubate the tissue slices as required. After the first antibody incubation is completed, suck out the incubation solution of the first antibody, and take out the incubation chamber inserted into the dovetail groove of the rectangular wall in the incubation frame to adjust the space. Block, remove the elastic holders on the four sides of the structure where the incubation frame and the glass slide overlap up and down, remove the incubation frame from the glass slide, and wash the tissue slices pasted on the glass slide; second antibody incubation is required , repeat the above operation.

The invention has a reasonable structure, and can tightly press the incubation frame on the glass slide pasted with tissue slices by using the elastic holder multiple times to form a non-leakage incubation chamber when performing immunohistochemical staining of tissues. And by inserting 1 to 4 incubation space adjustment blocks into the antibody incubation chamber to flexibly adjust the size of any side or multilateral space of the antibody incubation chamber, remove the excess space in the incubation chamber, and form an incubation space that is suitable and does not leak Incubation chamber. It is ensured that when antibody incubation is performed, the use of a smaller amount of antibody incubation solution can achieve the effect that all tissue sections pasted on the glass slide can be completely immersed in the antibody incubation solution, further saving the experimental cost of scientific research. At the same time, by taking out the space adjustment block of the incubation chamber and unloading the incubation frame several times, it is ensured that the experimental steps such as slide pretreatment, tissue mounting, washing, dehydration, transparency, and sealing are not affected by the incubation frame. All the experimental steps can be carried out conveniently and efficiently, and finally a good immunohistochemical staining experimental result can be obtained.

Description of drawings

The present invention will be further described below in conjunction with drawings and embodiments.

Fig. 1 is a three-dimensional schematic diagram of an incubation frame of a detachable slide incubator with adjustable volume.

Fig. 2 is a three-dimensional schematic diagram of a slide incubator that can be detachable and has an adjustable volume.

Fig. 3 is a three-dimensional schematic diagram of the elastic holder of the detachable slide incubator with adjustable volume.

Fig. 4 is a three-dimensional schematic diagram of an incubation chamber space adjustment block of a detachable slide incubator with adjustable volume.

Fig. 5 is a schematic diagram of a longitudinal section of a detachable and volume-adjustable slide incubator.

In the picture:

1. Stainless steel incubation frame: Pressed on the glass slide, together with the silica gel pad below it, an incubation chamber is formed from a rectangular space in the middle with the glass slide as the bottom surface.

2. Silicone pad of the stainless steel incubation frame: it has the same middle rectangular space as the stainless steel incubation frame, and is adhered to the stainless steel incubation frame. When the incubation chamber is formed, it is in contact with the glass slide to prevent the leakage of the incubation solution.

3. The dovetail groove on the rectangular wall in the incubation frame: fix the space adjustment block of the incubation room to prevent the space adjustment block of the incubation room from wandering in the incubation chamber.

4. Glass slide: Carries tissue slices and serves as the bottom surface of the incubation chamber.

5. The root of the elastic holder: connect to the clamping part of the elastic holder, and provide the holding part of the elastic holder when forming and disassembling the slide incubator.

6. Clamping part of the elastic holder: press the incubation frame and slide glass together to form an incubation chamber.

7. The clamping port of the elastic holder: the elastic holder is clamped on the incubation frame and the glass slide through the clamping port.

8. The space adjustment part of the incubation room space adjustment block: put it into the incubation chamber to adjust the size of the incubation chamber space.

9. The dovetail-shaped convex body of the space adjustment block of the incubation chamber: insert it into the dovetail groove on the inner wall of the incubation frame to prevent the space adjustment block of the incubation chamber from wandering in the incubation chamber.

10. Incubation chamber: A small space is formed to store the incubation solution, so as to not only use a small amount of incubation solution, but also ensure that the tissue sections on the glass slide are always immersed in the incubation solution.

11. Elastic holder.

detailed description

A detachable and volume-adjustable glass slide incubator, including a stainless steel incubation frame, which is pressed on the glass slide, and together with the silica gel pad below it, forms an incubation chamber with a rectangular space in the middle and a glass slide as the bottom surface ; The silica gel pad has the same middle rectangular space as the stainless steel incubation frame, and is adhered to the stainless steel incubation frame to contact with the glass slide to prevent the leakage of the incubation solution; the outer circumference of the stainless steel incubation frame and the outer circumference of the silica gel pad The size is the same as the outer circumference of the glass slide; there is also an elastic holder for clamping the stainless steel incubation frame and the silica gel pad on the glass slide; the elastic holder includes the root of the elastic holder, the elastic holder The clamping part and the clamping mouth of the elastic holder; the root of the elastic holder is connected to the clamping part of the elastic holder, and the elastic holder is provided when the slide incubator is formed and the slide incubator is disassembled. The holding position of the elastic holder; clamp the elastic holder on the incubation frame and the glass slide through the clamping port of the elastic holder, and the clamping part of the elastic holder clamps the incubation frame and the glass slide together to form an incubation chamber A dovetail groove is set on the four sides of the inner wall of the stainless steel incubation frame and the silicone cushion, and the dovetail groove is plugged with the space adjustment block of the incubation room with a dovetail-shaped convex body to adjust the size of the incubation room space and prevent the space adjustment of the incubation room Blocks wander around in the incubation chamber.

Process the selected glass slides according to the requirements of immunohistochemistry to ensure that the tissue sections have a good sticking effect on the glass slides. Select the tissue required for the experiment, slice it, and attach it to a glass slide. The tissue sections attached to the glass slides were blocked before antibody incubation. After completing the above experimental steps, press the stainless steel incubation frame (1) and the silica gel cushion layer (2) on the glass slide with the tissue slice attached (4), to ensure that all tissue sections are incubated in the incubation frame and the glass slide. The bottom of the chamber (10). Select any one of the four sides where the incubation frame overlaps with the glass slide, hold the root of the elastic holder (5), and align the clamping port (7) of the elastic holder with the incubation frame to overlap the slide The structure in which the elastic holder is pushed into the incubator box and the glass slide overlaps up and down. Make sure that the clamping part (6) of the elastic clamp presses the incubation frame and slide tightly together. Repeat the above-mentioned action on the other three sides to clamp the elastic holder (11). Make sure the incubation frame is in tight contact with the slide to create a leak-tight incubation chamber. After forming a non-leaking incubation chamber, according to the position where the tissue section is attached on the glass slide, selectively insert 1 to 4 incubation chamber space adjustment blocks into the 4 dovetail grooves on the rectangular wall of the incubation frame. The space adjustment block is composed of the space adjustment part (8) of the incubation room space adjustment block and the dovetail-shaped convex body (9), so as to flexibly adjust the size of any side or multilateral space of the rectangle in the incubation frame. Covering the tissue sections attached to the bottom of the incubation chamber is standard. After adjusting the size of the incubation chamber space, add the incubation solution containing the specific primary antibody to the incubation chamber, making sure that the tissue slice pasted on the bottom of the incubation chamber can be soaked in the incubation solution containing the specific primary antibody. Incubate tissue sections as required. After the incubation with the primary antibody is completed, suck out the incubation solution containing the specific primary antibody, take out the space adjustment block of the incubation chamber inserted into the dovetail groove on the rectangular wall of the incubation frame, and remove the four sides of the structure that overlaps the incubation frame and the glass slide. Elastic grippers to remove the incubation frame from the slide. Wash the tissue sections pasted on glass slides routinely. When it is necessary to incubate with the second antibody, repeat the above operations to form an incubation chamber with a suitable size and no leakage, add the incubation solution containing the specific second antibody, and incubate the tissue slices. After the incubation is complete, remove the incubation solution containing the specific secondary antibody. Repeat the above operation to disassemble the lower incubation frame. Post-processing such as washing, dehydration, transparency, and sealing of tissue sections pasted on glass slides. After all treatments were completed, the results of immunohistochemical staining of tissue sections were observed under a microscope.

Claims (3)

1. the using method of the detachable also microslide couveuse of volume adjustable, it is characterized in that: the described detachable also microslide couveuse of volume adjustable comprises stainless steel and hatches frame, stainless steel is hatched frame and is pressed on microslide, is that bottom surface is formed and hatches cell jointly with the silica gel bed course below it by the coffin of centre with microslide; Described silica gel bed course has hatches the identical intermediate rectangular space of frame with stainless steel, and sticks to stainless steel and hatch on frame, contacts, prevent the leakage of Incubating Solution with microslide; The peripheral dimension that stainless steel hatches the peripheral dimension of frame and silica gel bed course is all identical with microslide peripheral dimension; Separately be provided with and stainless steel hatched frame and silica gel bed course and be clamped in resilient clamp on microslide; Described resilient clamp comprises resilient clamp root, the clamping part of resilient clamp and the clamp port of resilient clamp; Resilient clamp root connects resilient clamp clamping part, and provides the hand position of resilient clamp when forming microslide couveuse and the couveuse operation of dismounting microslide; Being clamped in by resilient clamp by the clamp port of resilient clamp hatches on frame and microslide, and the clamping part of resilient clamp will be hatched the formation that is clamped together of frame and microslide and be hatched cell; The inwall four sides that stainless steel hatches frame and silica gel bed course arranges a dovetail groove respectively, dovetail groove coordinates with the camera incubata Space adjustment block grafting with swallow-tail form convex body, regulate the size in camera incubata space, prevent camera incubata Space adjustment block from hatching little indoor migration;

Using method is: process the microslide selected, to ensure that histotomy has good sticking effect on microslide according to immunohistochemical requirement; Choose the required tissue of experiment to cut into slices, and be attached on microslide; The histotomy be attached on microslide is carried out the Seal treatment before antibody incubation; After completing above-mentioned experimental procedure, stainless steel is hatched frame (1) and silica gel bed course (2) to be pressed in and to be pasted with (4) on the microslide of histotomy, guarantee the bottom of hatching cell (10) that all histotomies are all being hatched frame and microslide and formed; Select to hatch frame and any one side in microslide up and down overlapping four limits, hand-held resilient clamp root (5), the clamp port (7) of resilient clamp is aimed at the structure of hatching frame and microslide and being stacked up and down, resilient clamp is pushed the structure of hatching frame and microslide and being stacked up and down, ensure that the clamping part (6) of resilient clamp will hatch frame and microslide forces together tightly, its excess-three limit is repeated above-mentioned action and is sandwiched by resilient clamp (11), guarantee to hatch frame and microslide close contact, what formation was not leaked hatches cell; Formed do not leak hatch cell after, according to the position that histotomy attaches on microslide, optionally in 4 dovetail grooves of hatching in frame in rectangular wall, insert 1 to 4 camera incubata Space adjustment block, camera incubata Space adjustment block is made up of the Space adjustment part (8) of camera incubata Space adjustment block and swallow-tail form convex body (9), hatch rectangle in frame arbitrarily or the size in polygon space on one side with flexible, the camera incubata Space adjustment block of insertion is attached at histotomy bottom camera incubata for standard not hide; After regulating the size of hatching compartment spaces, to hatching in cell the Incubating Solution added containing special first antibody, be sure oing to be pasted onto the histotomy of hatching bottom cell and can be immersed in the Incubating Solution containing special first antibody; Carry out hatching of histotomy on request; After first antibody has been hatched, sucking-off is containing the Incubating Solution of special first antibody, take out the camera incubata Space adjustment block inserting and hatch in frame in rectangular wall dovetail groove, unload the resilient clamp of hatching structure four limit that frame and microslide are stacked up and down, unload from microslide and hatch frame, routinely carrying out washing treatment is carried out to the histotomy be pasted onto on microslide; Need to carry out second antibody when hatching, repeat aforesaid operations, again formed camera incubata space size suitable do not leak hatch cell, add the Incubating Solution containing special second antibody, histotomy hatched; After having hatched, remove containing special second antibody Incubating Solution; Hatch frame under repeating aforesaid operations dismounting, the histotomy be pasted onto on microslide is washed, dewaters, transparent, mounting aftertreatment; After all process complete, examine under a microscope the immunohistochemical staining result of histotomy.

2. the using method of the detachable also microslide couveuse of volume adjustable according to claim 1, is characterized in that: the Space adjustment part of camera incubata Space adjustment block: put into and hatch cell, regulates the size in camera incubata space.

3. the using method of the detachable also microslide couveuse of volume adjustable according to claim 1, it is characterized in that: the swallow-tail form convex body of camera incubata Space adjustment block: insert in the dovetail groove of hatching on frame inwall, prevent camera incubata Space adjustment block from hatching little indoor migration.