A dewaxing solution and dewaxing method suitable for immunohistochemical staining instrument
By using a dewaxing solution composed of fatty alcohol polyoxyethylene ether and ethanol, the cumbersome and harmful problems of traditional dewaxing methods are solved, and a non-toxic, odorless and pollution-free dewaxing effect is achieved. It replaces xylene and is suitable for the immune group. Descaling dyer.
Patent Information
- Application Number
- CN202410694432.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-05-31
- Publication Date
- 2024-12-10
- Estimated Expiration
- 2044-05-31
AI Technical Summary
The traditional dewaxing method is cumbersome and takes a long time to operate. It uses carcinogenic xylene solvents, which pollutes the environment and affects the health of operators.
Dewaxing solution consisting of 60% to 80% fatty alcohol polyoxyethylene ether and 20% to 40% ethanol is dewaxed by soaking paraffin-embedded tissue section samples. The temperature and time can be adjusted. After dewaxing, use water, Rinse with ethanol or PBS and perform subsequent immunostaining directly.
It achieves a non-toxic, harmless and odorless dewaxing effect, replaces the traditional xylene dewaxing method, simplifies operation, is friendly to the environment, and has no effect in immunohistochemical staining.
Smart Images
Figure CN118258668B_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of immunoassay, and in particular relates to a dewaxing solution and a dewaxing method suitable for an immunohistochemical staining instrument. Background Art
[0002] At present, the early diagnosis of benign and malignant tumors in clinical practice mainly relies on the staining results of pathological sections. The diseased tissues or cells are made into sections or smears, and then observed under a microscope after staining to examine the lesions they have occurred, explore the causes and pathogenesis of the lesions, and the occurrence and development process of the lesions, and finally make a pathological diagnosis. The basic steps of pathological sections include sampling, fixation, dehydration, transparency, wax immersion, embedding, slicing (as well as pasting and baking), dewaxing, staining, re-dehydration, re-transparency and sealing. Among the above steps, dewaxing is a necessary process.
[0003] Paraffin is a mixture of solid higher alkanes, the main component of which is C n H 2n+2 , where n = 17 to 35, its main components are straight-chain alkanes, and a small amount of alkanes with individual branches and monocyclic cycloalkanes with long side chains; the main straight-chain alkanes are n-docosane (C 22 H 46 ) and n-octadecane (C 28 H 58 ). Paraffin wax is soluble in non-polar solvents such as gasoline, carbon disulfide, xylene, ether, benzene, chloroform, carbon tetrachloride, naphtha, etc., but insoluble in polar solvents such as water and methanol.
[0004] The traditional dewaxing method is to soak in xylene for 15 minutes three times, then wash with anhydrous ethanol for 5 minutes, 95% alcohol for 5 minutes, 80% alcohol for 5 minutes, and distilled water. The whole process is not only cumbersome and time-consuming, but also xylene as a dewaxing agent is carcinogenic, highly volatile, and has a pungent odor. Long-term dewaxing will not only pollute the environment, but also affect the health of operators. In view of this, it is necessary to improve the traditional dewaxing solution. Summary of the invention
[0005] In order to solve the shortcomings of the prior art, the purpose of the present invention is to provide a dewaxing solution and a dewaxing method suitable for an immunohistochemical staining instrument, which can dewax cleanly without affecting the subsequent immunohistochemical staining process, and does not use organic solvents such as xylene, and is non-toxic and harmless.
[0006] In order to achieve the above object, the present invention adopts the following technical solution:
[0007] A dewaxing solution suitable for a full-automatic immunohistochemical staining instrument is composed of 60% to 80% by volume of fatty alcohol polyoxyethylene ether and 20% to 40% by volume of ethanol.
[0008] The method for dewaxing paraffin-embedded tissue sections using a dewaxing solution comprises the following specific steps: immersing the paraffin-embedded tissue section sample in the dewaxing solution to dewax.
[0009] Preferably, the temperature of the dewaxing solution is 15-25°C, and the soaking time is 10-15 minutes.
[0010] Preferably, the temperature of the dewaxing solution is greater than 50° C., and the immersion time is 5 to 10 minutes.
[0011] Preferably, after dewaxing is completed, the residual dewaxing solution on the slice is directly rinsed with any one of water, ethanol or PBS, and subsequent immunostaining operations can be carried out.
[0012] The invention is beneficial in that the dewaxing solution is non-toxic and tasteless, is not easy to volatilize, has excellent dewaxing effect, can replace the traditional xylene dewaxing method for paraffin-embedded tissue section samples, and is environmentally friendly. BRIEF DESCRIPTION OF THE DRAWINGS
[0013] Figure 1 is a staining effect diagram of the paraffin-embedded colon cancer tissue section sample in Example 1;
[0014] Figure 2 This is a staining effect diagram of the paraffin-embedded colon cancer tissue section sample in Comparative Example 1;
[0015] Figure 3 is a staining effect diagram of the paraffin-embedded tonsil tissue section sample in Example 2;
[0016] Figure 4 This is a staining effect diagram of the paraffin-embedded myeloma tissue section sample in Example 3. DETAILED DESCRIPTION
[0017] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.
[0018] Example 1: A dewaxing solution suitable for a fully automatic immunohistochemical staining instrument, comprising 60% by volume of fatty alcohol polyoxyethylene ether and the remainder being ethanol. The dewaxing method comprises the following specific steps:
[0019] Add paraffin-embedded colon cancer tissue sections to the dewaxing solution and soak at 60°C for 5-10 minutes. After dewaxing is completed, rinse the remaining dewaxing solution on the sections with water, and then stain the tissue sections according to the conventional immunostaining process. The staining results are as follows: Figure 1 shown.
[0020] Example 2: A dewaxing solution suitable for a fully automatic immunohistochemical staining instrument, comprising 80% by volume of fatty alcohol polyoxyethylene ether and the remainder being ethanol. The dewaxing method comprises the following specific steps:
[0021] Add the paraffin-embedded tonsil tissue sections to the dewaxing solution and soak for 10 to 15 minutes at 15 to 25°C. After dewaxing is completed, rinse the remaining dewaxing solution on the sections with ethanol, and then stain the tissue sections according to the conventional immunostaining process. The staining results are as follows: Figure 3 shown.
[0022] Example 3: A dewaxing solution suitable for a fully automatic immunohistochemical staining instrument, comprising 70% by volume of fatty alcohol polyoxyethylene ether and the remainder being ethanol. The dewaxing method comprises the following specific steps:
[0023] Add paraffin-embedded myeloma tissue sections to the dewaxing solution and soak at 72°C for 5 to 10 minutes. After dewaxing is completed, rinse the remaining dewaxing solution on the sections with PBS, and then stain the tissue sections according to the conventional immunostaining process. The staining results are as follows: Figure 4 shown.
[0024] Comparative Example 1: The same paraffin-embedded colon cancer tissue sections as in Example 1 were dewaxed using a conventional dewaxing method, specifically in the following steps:
[0025] Paraffin-embedded colon cancer tissue sections were dewaxed in xylene for 15 minutes three times, washed with anhydrous ethanol for 5 minutes three times, then washed with 95% alcohol for 5 minutes, 80% alcohol for 5 minutes, and 70% alcohol for 5 minutes, and finally rinsed with distilled water; then the tissue section samples were stained according to the conventional immunostaining process. The staining results are shown in the figure. Figure 2 shown.
[0026] pass Figure 1 and Figure 2 It can be seen that compared with the traditional xylene dewaxing method, the dewaxing method in Example 1 has no difference in effect with the xylene dewaxing method. Therefore, the dewaxing liquid and dewaxing method of the present invention can completely replace the traditional dewaxing method.
[0027] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the above embodiments do not limit the present invention in any form, and any technical solution obtained by equivalent replacement or equivalent transformation falls within the protection scope of the present invention.
Claims
1. A dewaxing solution suitable for a fully automatic immunohistochemical staining instrument, characterized in that: It is composed of 60% to 80% by volume of fatty alcohol polyoxyethylene ether and 20% to 40% by volume of ethanol.
2. A method for dewaxing paraffin-embedded tissue sections using the dewaxing solution according to claim 1, characterized in that: The specific steps are as follows: the paraffin-embedded tissue section sample is immersed in a dewaxing solution to dewax.
3. The method for dewaxing paraffin-embedded tissue sections according to claim 2, characterized in that: The temperature of the dewaxing solution is 15-25° C., and the soaking time is 10-15 minutes.
4. The method for dewaxing paraffin-embedded tissue sections according to claim 2, characterized in that: The temperature of the dewaxing solution is 60° C. or 72° C., and the soaking time is 5 to 10 minutes.