CN113686634A - Dewaxing repair liquid beneficial to acquisition of paraffin section images - Google Patents
Dewaxing repair liquid beneficial to acquisition of paraffin section images Download PDFInfo
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- CN113686634A CN113686634A CN202110914714.3A CN202110914714A CN113686634A CN 113686634 A CN113686634 A CN 113686634A CN 202110914714 A CN202110914714 A CN 202110914714A CN 113686634 A CN113686634 A CN 113686634A
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Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
Abstract
The invention relates to a dewaxing repair liquid beneficial to obtaining a paraffin section image, which comprises the following dewaxing steps: the tissue paraffin section is placed in an oven at 60-65 ℃ for baking for 30-60 minutes, and then the section is placed in a dewaxing repair liquid preheated to 72-75 ℃, the dewaxing repair liquid integrates the operation steps of section dewaxing, hydration and antigen repair, the antigen repair is carried out while dewaxing, hydration and repair are completed at one time, the operation steps are shortened, the experimental time is saved, the dewaxing repair liquid has the advantages that the dewaxing repair liquid is nontoxic and difficult to volatilize, a defoaming agent is added in the operation process, the volatilization of a liquid reagent can be effectively prevented under the heating state to generate irritant gas, meanwhile, the influence of bubbles formed on the surface of a glass slide on the dewaxing repair effect is prevented, the biodegradability is good, the dewaxing effect is equal to that of xylene, and the pathological section treated by the reagent has good dewaxing effect, and does not damage the cell structure of the pathological tissue.
Description
Technical Field
The invention relates to the technical field of paraffin section, in particular to a dewaxing repair liquid beneficial to acquisition of paraffin section images.
Background
Pathological paraffin tissue sections are the most common and the most main detection samples for Immunohistochemistry (IHC); the sample is a paraffin-embedded tissue section, is water-insoluble and cannot be directly used for immunohistochemical detection, and the sample needs to be pretreated before immunohistochemical detection, namely, the sample needs to be dewaxed, hydrated and antigen-repaired;
the typical paraffin tissue section pretreatment basic steps are as follows: dewaxing xylene (or an environment-friendly dewaxing agent), performing ethanol gradient hydration (such as absolute ethyl alcohol, 95% ethanol, 85% ethanol and distilled water), repairing antigens, and rinsing; after a series of processes, antigen-antibody immunoreaction dyeing can be carried out; the classical paraffin tissue section dewaxing, hydrating and repairing method has the advantages that not only are the operation steps complex, but also xylene is often used for dewaxing, has certain toxicity and is not environment-friendly, and certain damage can be caused to the operation.
Disclosure of Invention
The invention aims to provide a dewaxing repair liquid beneficial to obtaining a paraffin section image, and solves the problems.
The invention realizes the purpose through the following technical scheme: the dewaxing repair liquid beneficial to paraffin section image acquisition is characterized by comprising the following dewaxing steps: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 30-60 minutes, then placing the section in a dewaxing repair liquid preheated to 72-75 ℃, and soaking for 20 minutes at constant temperature; taking out the slices, directly putting the slices into another cylinder, preheating the slices to 72-75 ℃, soaking the slices in the dewaxing repair liquid for 20 minutes at constant temperature, and then washing the slices with water to finish dewaxing.
Preferably, the method comprises the following steps of dewaxing and antigen retrieval: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 30-60 minutes, then placing the section in the dewaxing repair liquid which is preheated to 72-75 ℃, and continuously heating the dewaxing repair liquid to 95-100 ℃ for 20 minutes; and after the slices are taken out, rinsing the slices once in the dewaxing repair liquid preheated to 72-75 ℃, and then washing the slices with water to finish dewaxing repair.
Preferably, dewaxing is accomplished by incubation at 72-75 ℃.
Preferably, the dewaxing and antigen retrieval are accomplished in one step at a temperature of 95-100 ℃.
Preferably, the dewaxing agent contained in the dewaxing repair liquid comprises: the paraffin wax remover comprises a water-soluble dewaxing agent, a penetrating agent, a dispersing agent, a defoaming agent, distilled water and the like, wherein the water-soluble dewaxing agent is a mixture consisting of PEG200, glycol and fatty alcohol polyoxyethylene 9 ether, has good solubility in a water phase, and can dissolve paraffin wax under the condition of heating; the penetrant is compound of two or at least one of Triton X-100 and glycerol; the dispersant is a surfactant and is at least one compound of Tween-20 and Triton X-100; the defoaming agent is two or at least one compound of ethylene glycol tertiary butyl ether and polyoxypropylene glycerol ether.
Preferably, the water-soluble dewaxing agent comprises the following components in percentage by weight: 0.25 to 2.5 percent of PEG200, 0.05 to 0.75 percent of glycol and 0.05 to 0.75 percent of fatty alcohol-polyoxyethylene ether; the optimal component ratio of the penetrating agent is as follows: 0.1% -0.75% of Triton X-100 and 0.01% -0.5% of glycerol; the optimal component proportion of the dispersant is 0.1 to 0.75 percent of Triton X-100 and 0.01 to 0.05 percent of Tween-20; the optimal component ratio of the defoaming agent is as follows: 0.01 to 0.1 percent of ethylene glycol tertiary butyl ether and 0.03 to 0.05 percent of polyoxypropylene glycerol ether.
Preferably, the repair liquid contained in the dewaxing repair liquid includes: the repairing solution comprises a buffer solution, a penetrating agent and a chelating agent, wherein the buffer solution, the penetrating agent and the chelating agent are main components in the repairing solution, are prepared from Tris hydrochloric acid, and have a pH value of 9.0; the penetrant can be Triton X-100, glycerol, or two or at least one compound; the chelating agent, EDTA, can reduce the interference of metal.
Preferably, the buffer solution comprises the following components in percentage by weight: 0.01-0.05mM Tris-HCl; the optimal component content of the metal chelating agent is as follows: 0.001-0.002mM EDTA; the optimal component content of the tissue penetrating agent is 0.01-0.05% (V/V) of glycerol.
Preferably, when the dewaxing repair liquid is used in a pure dewaxing operation, the temperature for heating dewaxing incubation is 72-75 ℃.
Preferably, when the dewaxing repair liquid is used in a single dewaxing operation, the dewaxing repair liquid is heated, dewaxed and incubated for 20 minutes and is soaked twice at constant temperature.
In the present invention, preferably, when the dewaxing repair liquid is used in a single dewaxing operation, the rinse water is tap water after completion of heating dewaxing.
Preferably, during the dewaxing repair operation, the dewaxing repair liquid needs to be heated to 72-75 ℃ in advance, and the paraffin tissue section cannot be directly put into the cold dewaxing repair liquid, so that paraffin on the section is prevented from adhering to the glass slide and is not beneficial to the diffusion of the dissolved wax and the wax.
Preferably, the dewaxing repair liquid is put into the paraffin tissue section and then is continuously heated to 95-100 ℃ during the dewaxing repair operation, so that the dewaxing can be promoted and the purpose of antigen repair can be achieved under the temperature condition.
In the present invention, preferably, the dewaxing repair liquid is placed in the paraffin tissue section during the dewaxing repair operation, and then the dewaxing repair liquid is heated for dewaxing repair, wherein the duration of the heating is 20 minutes.
Preferably, the paraffin tissue section subjected to dewaxing repair is taken out and rinsed once by the dewaxing repair liquid preheated to 72-75 ℃ before being flushed by tap water.
Compared with the prior art, the invention has the following beneficial effects: the dewaxing repair liquid integrates the operation steps of section dewaxing, hydration and antigen repair, carries out antigen repair while dewaxing, finishes dewaxing-hydration-repair at one time, shortens the operation steps, saves the experimental time, and has the advantages that the dewaxing repair liquid is non-toxic and not easy to volatilize, and can effectively prevent the volatilization of a liquid reagent to generate irritant gas under the heating state due to the addition of a defoaming agent in the operation process, and simultaneously prevent the dewaxing repair effect from being influenced by bubbles formed on the surface of a glass slide, the biodegradability is good, the dewaxing effect is equal to that of xylene, and pathological sections treated by the reagent have good dewaxing effect and cannot damage the cell structure of pathological tissues.
Drawings
FIG. 1 is a graph showing the results of HE staining of a dewaxing repair liquid according to a first embodiment of the present invention;
FIG. 2 is a schematic diagram of the IHC staining result of the first embodiment of the present invention;
FIG. 3 is a graph showing the results of HE staining of a dewaxing repair liquid according to example II of the present invention;
FIG. 4 is a graph showing the result of IHC staining in the second embodiment of the present invention;
FIG. 5 is a graph showing the results of HE staining of a dewaxing repair liquid according to example III of the present invention;
FIG. 6 is a graph showing the result of IHC staining in the third embodiment of the present invention.
Detailed Description
The invention will be further described with reference to the accompanying drawings in which:
a dewaxing repair liquid for facilitating the acquisition of paraffin section images, as shown in fig. 1-6, comprising the following dewaxing steps: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 30-60 minutes, then placing the section in a dewaxing repair liquid preheated to 72-75 ℃, and soaking for 20 minutes at constant temperature; taking out the slices, directly putting the slices into another cylinder, preheating the slices to 72-75 ℃, soaking the slices in the dewaxing repair liquid for 20 minutes at constant temperature, and then washing the slices with water to finish dewaxing.
Further, the method comprises the following steps of dewaxing and antigen retrieval: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 30-60 minutes, then placing the section in the dewaxing repair liquid which is preheated to 72-75 ℃, and continuously heating the dewaxing repair liquid to 95-100 ℃ for 20 minutes; and after the slices are taken out, rinsing the slices once in the dewaxing repair liquid preheated to 72-75 ℃, and then washing the slices with water to finish dewaxing repair.
Further, dewaxing is accomplished by incubation at 72-75 ℃.
Further, under the condition of heating at 95-100 ℃, the dewaxing and antigen retrieval effects are completed at one time.
Further, the dewaxing repair liquid contains dewaxing agents comprising: the paraffin wax remover comprises a water-soluble dewaxing agent, a penetrating agent, a dispersing agent, a defoaming agent, distilled water and the like, wherein the water-soluble dewaxing agent is a mixture consisting of PEG200, glycol and fatty alcohol polyoxyethylene 9 ether, has good solubility in a water phase, and can dissolve paraffin wax under the condition of heating; the penetrant is compound of two or at least one of Triton X-100 and glycerol; the dispersant is a surfactant and is at least one compound of Tween-20 and Triton X-100; the defoaming agent is two or at least one compound of ethylene glycol tertiary butyl ether and polyoxypropylene glycerol ether.
Further, the optimal component ratio of the water-soluble dewaxing agent is as follows: 0.25 to 2.5 percent of PEG200, 0.05 to 0.75 percent of glycol and 0.05 to 0.75 percent of fatty alcohol-polyoxyethylene ether; the optimal component ratio of the penetrating agent is as follows: 0.1% -0.75% of Triton X-100 and 0.01% -0.5% of glycerol; the optimal component proportion of the dispersant is 0.1 to 0.75 percent of Triton X-100 and 0.01 to 0.05 percent of Tween-20; the optimal component ratio of the defoaming agent is as follows: 0.01 to 0.1 percent of ethylene glycol tertiary butyl ether and 0.03 to 0.05 percent of polyoxypropylene glycerol ether.
Further, the repair liquid contained in the dewaxing repair liquid comprises: the repairing solution comprises a buffer solution, a penetrating agent and a chelating agent, wherein the buffer solution, the penetrating agent and the chelating agent are main components in the repairing solution, are prepared from Tris hydrochloric acid, and have a pH value of 9.0; the penetrant can be Triton X-100, glycerol, or two or at least one compound; the chelating agent, EDTA, can reduce the interference of metal.
Further, the optimal component content of the buffer solution is as follows: 0.01-0.05mM Tris-HCl; the optimal component content of the metal chelating agent is as follows: 0.001-0.002mM EDTA; the optimal component content of the tissue penetrating agent is 0.01-0.05% (V/V) of glycerol.
Further, when the dewaxing repair liquid is used for a pure dewaxing operation, the heating dewaxing incubation temperature is 72-75 ℃.
Further, when the dewaxing repair liquid is used for a pure dewaxing operation, the heating dewaxing incubation time is 20 minutes, and the dewaxing repair liquid is soaked twice at constant temperature.
Furthermore, when the dewaxing repair liquid is used for pure dewaxing operation, the rinsing water is tap water after heating dewaxing is finished.
Furthermore, when the dewaxing repair liquid is used for dewaxing repair operation, the dewaxing repair liquid needs to be heated to 72-75 ℃ in advance, and the paraffin tissue section cannot be directly placed into the cold dewaxing repair liquid, so that paraffin on the section is prevented from being adhered to the glass slide, and the paraffin is prevented from being diffused by the paraffin.
Furthermore, when the dewaxing repair liquid is used for dewaxing repair operation, the dewaxing repair liquid is required to be continuously heated to 95-100 ℃ after being placed into a paraffin tissue section, and the dewaxing can be promoted and the purpose of antigen repair can be achieved under the temperature condition.
Furthermore, during the dewaxing repair operation, the dewaxing repair liquid needs to be heated and dewaxed continuously after being placed into the paraffin tissue section, and the duration time needs to be 20 minutes.
Furthermore, when the dewaxing repair liquid is used for dewaxing repair operation, the paraffin tissue section subjected to dewaxing repair is taken out and rinsed once by the dewaxing repair liquid preheated to 72-75 ℃, and then the paraffin tissue section can be flushed by tap water.
The first embodiment,
The preparation method of the dewaxing repair liquid for paraffin tissue sections and the application thereof are as follows:
preparation operation step 1: preparing 0.01mM Tris-HCl (pH9.0) buffer solution, and adding EDTA to make its concentration reach 0.001mM, thus obtaining the antigen retrieval solution. Weighing 1.21g of Tris and 1.29g of EDTA0, dissolving in 1L of distilled water, and adjusting the pH value to 9.0 by using hydrochloric acid after full dissolution;
preparation operation step 2: according to the composition proportion, sequentially weighing a water-soluble dewaxing agent (PEG 200, ethylene glycol, fatty alcohol polyoxyethylene ether), a penetrating agent (Triton X-100, glycerol), a dispersing agent (Tween-20) and a defoaming agent (ethylene glycol tert-butyl ether), adding into the antigen retrieval liquid obtained in the step (1), and fully stirring and dissolving;
this example is a preparation and application of a dewaxing repair liquid for paraffin tissue sections, wherein the dewaxing repair liquid serves as a dewaxing agent (hydration) in an immunohistochemical experiment: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 60 minutes, then placing the section in dewaxing repair liquid preheated to 75 ℃, and soaking for 20 minutes at constant temperature; taking out the slices, directly putting the slices into another cylinder, preheating the slices to 75 ℃ in advance, soaking the slices for 20 minutes at constant temperature, and then washing the slices with water to finish dewaxing and hydration. Dewaxing the hydrated pathological section, performing HE staining, dehydrating, transparentizing and sealing;
fig. 1 shows the results of HE staining of the dewaxing repair liquid of this example: the result shows that no wax is left on the surface of the section, the dewaxing is clean, the tissue cell morphology is complete, the dyeing is clear and obvious, and the pathological section treated by the reagent has good dewaxing effect and the cell structure of the pathological tissue cannot be damaged;
the present embodiment relates to a preparation method and an application of a dewaxing repair liquid for paraffin tissue sections, wherein the dewaxing repair liquid has dewaxing, hydration and antigen repair effects in an immunohistochemical experiment: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 60 minutes, then placing the section in a dewaxing repair liquid preheated to 75 ℃, and continuously heating the dewaxing repair liquid to 100 ℃ for 20 minutes; taking out the slices, rinsing the slices once in dewaxing repair liquid preheated to 75 ℃, and then washing the slices with water to finish dewaxing repair;
fig. 2 shows the IHC staining results of the dewaxing repair liquid of this example: the staining result of TTF-1 on lung cancer tissues shows that the wax on the surfaces of the sections is not obviously retained, the dewaxing effect is obviously clean, good antigen repair can be seen through TTF-1 staining, the tissue cell morphology is kept in a good original state and the integrity thereof, cell nucleus hematoxylin negatively expressed by TTF-1 is clearly and obviously stained, and a TTF-1 positive staining signal is clear and definite. The dewaxing repair liquid meets the requirements of immunohistochemical dewaxing, hydration and antigen repair of tissue slices.
Example II,
The preparation method of the dewaxing repair liquid for paraffin tissue sections and the application thereof are as follows:
preparation operation step 1: preparing 0.01mM Tris-HCl (pH9.0) buffer solution, and adding EDTA to make its concentration reach 0.001mM, thus obtaining the antigen retrieval solution. Weighing 1.21g of Tris and 1.29g of EDTA0, dissolving in 1L of distilled water, and adjusting the pH value to 9.0 by using hydrochloric acid after full dissolution;
preparation operation step 2: according to the composition proportion, sequentially weighing a water-soluble dewaxing agent (PEG 200, ethylene glycol, fatty alcohol polyoxyethylene ether), a penetrating agent (Triton X-100, glycerol), a dispersing agent (Tween-20) and a defoaming agent (polyoxypropylene glycerol ether), adding the water-soluble dewaxing agent, the penetrating agent (Triton X-100, glycerol), the dispersing agent and the defoaming agent into the antigen repairing solution obtained in the step (1), and fully stirring and dissolving the mixture;
in the preparation and application of the dewaxing repair liquid for paraffin tissue sections, the dewaxing repair liquid serves as a dewaxing agent (hydration) in an immunohistochemical experiment: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 60 minutes, then placing the section in dewaxing repair liquid preheated to 75 ℃, and soaking for 20 minutes at constant temperature; taking out the slices, directly putting the slices into another cylinder, preheating the slices to 75 ℃ in advance, soaking the slices for 20 minutes at constant temperature, and then washing the slices with water to finish dewaxing and hydration. Dewaxing the hydrated pathological section, performing HE staining, dehydrating, transparentizing and sealing;
fig. 3 shows the results of HE staining of the dewaxing repair liquid of this example: the result shows that no wax is left on the surface of the section, the dewaxing is clean, the tissue cell morphology is complete, the dyeing is clear and obvious, and the pathological section treated by the reagent has good dewaxing effect and the cell structure of the pathological tissue cannot be damaged;
the preparation and application of the dewaxing repair liquid for paraffin tissue sections of the embodiment include dewaxing, hydration and antigen repair effects of the dewaxing repair liquid in immunohistochemical experiments: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 60 minutes, then placing the section in a dewaxing repair liquid preheated to 75 ℃, and continuously heating the dewaxing repair liquid to 100 ℃ for 20 minutes; taking out the slices, rinsing the slices once in dewaxing repair liquid preheated to 75 ℃, and then washing the slices with water to finish dewaxing repair;
fig. 4 shows the IHC staining results of the dewaxing repair liquid of this example: the result of CK5/6 staining on tonsil tissues shows that the wax on the surface of the section is not obviously retained, the dewaxing effect is obviously clean, good antigen repair can be seen through CK5/6 staining, the form of tissue cells is kept in a good original state and the integrity of the tissue cells, CK5/6 negative expression karyophilin staining is clear and obvious, and CK5/6 positive staining signals are clear and definite. The dewaxing repair liquid meets the requirements of immunohistochemical dewaxing, hydration and antigen repair of tissue slices.
Example III,
The preparation method of the dewaxing repair liquid for paraffin tissue sections and the application thereof are as follows:
preparation operation step 1: preparing 0.01mM Tris-HCl (pH9.0) buffer solution, and adding EDTA to make its concentration reach 0.001mM, thus obtaining the antigen retrieval solution. Weighing 1.21g of Tris and 1.29g of EDTA0, dissolving in 1L of distilled water, and adjusting the pH value to 9.0 by using hydrochloric acid after full dissolution;
preparation operation step 2: according to the composition proportion, sequentially weighing a water-soluble dewaxing agent (PEG 200, ethylene glycol, fatty alcohol polyoxyethylene ether), a penetrating agent (Triton X-100, glycerol), a dispersing agent (Tween-20) and a defoaming agent (polyoxypropylene glycerol ether), adding the water-soluble dewaxing agent, the penetrating agent (Triton X-100, glycerol), the dispersing agent and the defoaming agent into the antigen repairing solution obtained in the step (1), and fully stirring and dissolving the mixture;
in the preparation and application of the dewaxing repair liquid for paraffin tissue sections, the dewaxing repair liquid serves as a dewaxing agent (hydration) in an immunohistochemical experiment: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 60 minutes, then placing the section in dewaxing repair liquid preheated to 75 ℃, and soaking for 20 minutes at constant temperature; taking out the slices, directly putting the slices into another cylinder, preheating the slices to 75 ℃ in advance, soaking the slices for 20 minutes at constant temperature, and then washing the slices with water to finish dewaxing and hydration. Dewaxing the hydrated pathological section, performing HE staining, dehydrating, transparentizing and sealing;
fig. 5 shows the results of HE staining of the dewaxing repair liquid of this example: the result shows that no wax is left on the surface of the section, the dewaxing is clean, the tissue cell morphology is complete, the dyeing is clear and obvious, and the pathological section treated by the reagent has good dewaxing effect and the cell structure of the pathological tissue cannot be damaged;
the preparation and application of the dewaxing repair liquid for paraffin tissue sections of the embodiment include dewaxing, hydration and antigen repair effects of the dewaxing repair liquid in immunohistochemical experiments: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 60 minutes, then placing the section in a dewaxing repair liquid preheated to 75 ℃, and continuously heating the dewaxing repair liquid to 100 ℃ for 20 minutes; taking out the slices, rinsing the slices once in dewaxing repair liquid preheated to 75 ℃, and then washing the slices with water to finish dewaxing repair;
fig. 6 shows the IHC staining results of the dewaxing repair liquid of this example: the staining result of p63 on tonsil tissue shows that the wax on the surface of the section is not obviously retained, the dewaxing effect is obviously clean, the antigen repair is good through the p63 staining, the histocyte morphology is kept in a good original state and the integrity, the staining of the p63 negative expression cell nucleus hematoxylin is clear and obvious, and the p63 positive staining signal is clear and definite. The dewaxing repair liquid meets the requirements of immunohistochemical dewaxing, hydration and antigen repair of tissue slices.
The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed.
Claims (14)
1. The dewaxing repair liquid beneficial to paraffin section image acquisition is characterized by comprising the following dewaxing steps: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 30-60 minutes, then placing the section in a dewaxing repair liquid preheated to 72-75 ℃, and soaking for 20 minutes at constant temperature; taking out the slices, directly putting the slices into another cylinder, preheating the slices to 72-75 ℃, soaking the slices in the dewaxing repair liquid for 20 minutes at constant temperature, and then washing the slices with water to finish dewaxing.
2. The dewaxing repair liquid for facilitating paraffin section image acquisition as claimed in claim 1, comprising the following dewaxing and antigen repairing steps: placing the tissue paraffin section in an oven at 60-65 ℃ for baking for 30-60 minutes, then placing the section in the dewaxing repair liquid which is preheated to 72-75 ℃, and continuously heating the dewaxing repair liquid to 95-100 ℃ for 20 minutes; and after the slices are taken out, rinsing the slices once in the dewaxing repair liquid preheated to 72-75 ℃, and then washing the slices with water to finish dewaxing repair.
3. The fluid for restoring dewaxing of claim 1, wherein dewaxing is accomplished by incubation at 72-75 ℃.
4. The dewaxing repair liquid facilitating paraffin section image acquisition as claimed in claim 2, wherein dewaxing and antigen repair are completed at one time under the condition of heating at 95-100 ℃.
5. The dewaxing repair liquid facilitating paraffin section image acquisition is characterized in that a dewaxing agent contained in the dewaxing repair liquid comprises: water-soluble dewaxing agent, penetrating agent, dispersing agent, defoaming agent, distilled water and the like.
6. The dewaxing repair liquid beneficial to paraffin section image acquisition according to claim 5, wherein the optimal component ratio of the water-soluble dewaxing agent is as follows: 0.25 to 2.5 percent of PEG200, 0.05 to 0.75 percent of glycol and 0.05 to 0.75 percent of fatty alcohol-polyoxyethylene ether; the optimal component ratio of the penetrating agent is as follows: 0.1% -0.75% of Triton X-100 and 0.01% -0.5% of glycerol; the optimal component proportion of the dispersant is 0.1 to 0.75 percent of Triton X-100 and 0.01 to 0.05 percent of Tween-20; the optimal component ratio of the defoaming agent is as follows: 0.01 to 0.1 percent of ethylene glycol tertiary butyl ether and 0.03 to 0.05 percent of polyoxypropylene glycerol ether.
7. The dewaxing repair liquid for facilitating paraffin section image acquisition according to claim 5, wherein the repair liquid contained in the dewaxing repair liquid comprises: buffer solution, osmotic agent and chelating agent.
8. The dewaxing repair solution for facilitating paraffin section image acquisition according to claim 7, wherein the optimal component content of the buffer solution is as follows: 0.01-0.05mM Tris-HCl; the optimal component content of the metal chelating agent is as follows: 0.001-0.002mM EDTA; the optimal component content of the tissue penetrating agent is 0.01-0.05% (V/V) of glycerol.
9. The dewaxing repair liquid for facilitating paraffin section image acquisition according to claim 7, wherein the dewaxing repair liquid is used for a pure dewaxing operation, and the heating dewaxing incubation temperature of the dewaxing repair liquid is 72-75 ℃.
10. The dewaxing repair liquid for paraffin section image acquisition as claimed in claim 9, wherein the dewaxing repair liquid is used in a pure dewaxing operation, and is heated, dewaxed and incubated for 20 minutes, and soaked twice at constant temperature.
11. The fluid for restoring dewaxing of paraffin section image acquisition in accordance with claim 10, wherein the fluid for restoring dewaxing is used in a pure dewaxing operation, and the rinsing water is tap water after the heating dewaxing operation.
12. The fluid for restoring paraffin wax beneficial to image acquisition of paraffin wax sections according to claim 11, wherein the fluid for restoring paraffin wax requires preheating to 72-75 ℃ during the restoring dewaxing operation, and the paraffin wax tissue sections cannot be directly put into the cold fluid for restoring paraffin wax.
13. The fluid for restoring paraffin wax beneficial to the acquisition of paraffin wax section images as claimed in claim 12, wherein the fluid for restoring paraffin wax is further heated to 95-100 ℃ after being placed in paraffin wax tissue sections during the restoring paraffin wax operation.
14. The paraffin restoration solution facilitating paraffin section image acquisition as claimed in claim 13, wherein the paraffin restoration solution finishes the paraffin tissue section subjected to paraffin restoration during the paraffin restoration operation, and is taken out and rinsed once by the paraffin restoration solution preheated to 72-75 ℃ and then rinsed by tap water.
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| CN114486423A (en) * | 2021-12-29 | 2022-05-13 | 杭州迪英加科技有限公司 | Preparation method of compound liquid for paraffin tissue section dewaxing repair blocking |
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