A quick fixative for pathological specimen ink staining and its preparation method and application method

Abstract

The invention discloses a rapid fixing solution for ink staining of pathological specimens and a preparation method and an application method thereof, which comprises the following components by weight: 100 to 200 parts of glacial acetic acid, 200 to 400 parts of ethanol, 200 to 400 parts of methanol, 50 to 200 parts of tert-amyl alcohol, and 50 to 200 parts of formalin. The rapid fixing solution for ink staining of the invention can stain specimens rapidly and has a good staining effect, and the fixing solution of the invention does not need to use picric acid which is explosive and toxic.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, in particular to a quick fixative for pathological specimen ink staining and a preparation method and an application method thereof. Background Art

[0002] In clinical pathology, we often need to prepare pathological specimens as a basis for disease diagnosis. The cutting margin of pathological specimens is very important for judging whether many specimens (especially tumor tissue) are removed cleanly. The staining of cutting margins is very important for pathologists to judge whether the tissue is a surgical cutting margin and to judge the location and direction of the tissue under a microscope. At present, different colors of ink are often used to stain fresh specimens. After subsequent preparation, it is possible to observe at a glance under the microscope whether there is residual tumor tissue at the surgical cutting margin.

[0003] Ink staining of tissues that need to be stained, such as the cutting edge, is rarely used in primary hospitals. This is mainly due to two reasons: First, after fresh tissue specimens are stained with ink, if no fixative is used, it takes a very long time to dry, usually more than half an hour to wait, and whether the specimen is taken immediately after application or after drying, the ink will dissolve in the water, making the specimen table a mess. The final slices have very little ink left and are almost unrecognizable, which is not suitable for widespread clinical promotion. Second, the foreign practice is to use Bouin's solution (75% saturated aqueous solution of picric acid and 25% formalin added to deionized water and dripped on the ink for fixation, and then blotted with absorbent paper to achieve a rapid fixation effect. However, Bouin's solution contains picric acid, which is explosive and toxic, and is not suitable for clinical application.

[0004] In summary, the present invention provides a rapid fixative for ink staining of pathological specimens, which has better safety and clinical practicality compared with the prior art. The application of the present invention is of great significance to improving the development of pathology and the level of clinical pathology diagnosis and treatment in my country. Summary of the invention

[0005] In order to overcome the defects in the prior art, the present invention provides a rapid fixative for pathological specimens stained with Chinese ink and a preparation method and an application method thereof, which has the characteristics of low cost, easy configuration and convenient use, and the fixative of the present invention does not need to use explosive and toxic picric acid. The present invention discloses a rapid fixative for pathological specimens stained with Chinese ink, which comprises the following components by weight:

[0006] 100-200 parts of glacial acetic acid,

[0007] 200-400 parts of ethanol,

[0008] 200-400 parts of methanol,

[0009] 10 tert-amyl alcohol 50-200 parts,

[0010] 50-200 parts of formalin.

[0011] In some embodiments, a rapid fixative for staining pathological specimens with Indian ink comprises the following components in parts by weight:

[0012] 200 parts of glacial acetic acid,

[0013] 15 300 parts of ethanol,

[0014] 300 parts of methanol,

[0015] 100 parts of tert-amyl alcohol,

[0016] Formalin 100 parts.

[0017] The present invention also discloses a method for preparing a fixing liquid, comprising the following steps:

[0018] Step 1: Ultrasonically mix 100-200 parts of glacial acetic acid, 200-400 parts of ethanol, and 200-400 parts of methanol to obtain liquid A;

[0019] Step 2: Fully mix 50-200 parts of tert-amyl alcohol and 50-200 parts of formalin to obtain liquid B;

[0020] Step 3: Ultrasonic mixing of the solution A in step 1 and the solution B in step 2 is performed to obtain a stationary solution.

[0021] Optionally, in step 1, the ultrasonic power of the ultrasonic mixing is 100 W, the ultrasonic time is 10 to 15 min, and the mixing temperature is room temperature.

[0022] Optionally, the ultrasonic power of the ultrasonic mixing in step three is 200 W, the ultrasonic time is 20 to 25 min, and the mixing temperature is room temperature.

[0023] The present invention also discloses a method for fixing pathological specimens by ink staining, comprising the following steps:

[0024] Step 1: Take samples from the cut edge of the pathological specimen, stain with ink, and let stand for 30 seconds;

[0025] Step 2: Spray the fixative prepared above evenly on the sample stained in step 1 and let it stand for 30 seconds;

[0026] Step 3: Rinse the sample with water to wash away excess reagent after fixation.

[0027] Optionally, the pathological specimen size is 0.025cm2 to 4cm2.

[0028] In some embodiments, the pathological specimen is lung membrane, and the India ink staining and fixation method thereof comprises the following steps:

[0029] Step 1: Take a lung membrane sample from the cut edge of the lung tissue, stain it with ink, and let it stand for 30 seconds;

[0030] Step 2: Spray the fixative solution evenly on the sample stained in step 1 and let it stand for 30 seconds;

[0031] Step 3: Rinse the sample with water to wash away excess reagent after fixation. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0033] Figure 1 This is a picture showing the effect of dyeing and fixing using Example 1, which shows that the dyeing is uniform, there is less residual dye, and the effect is good.

[0034] Figure 2 This is a picture showing the effect of dyeing and fixing using Example 2, which shows that the dyeing is uniform, there is less residual dye, and the effect is good.

[0035] Figure 3 This is a picture showing the effect of dyeing and fixing using Example 3, which shows that the dyeing is uniform, there is less residual dye, and the effect is good.

[0036] Figure 4 This is a picture showing the effect of dyeing and fixing using Example 4. The picture shows that the dyeing is uneven, there is less residual dye, and the effect is average.

[0037] Figure 5 This is a picture showing the effect of dyeing and fixing using Comparative Example 1. The picture shows that the dyeing is uniform, there is a lot of residual dye, and the effect is average. DETAILED DESCRIPTION

[0038] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0039] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0040] Example 1 Preparation of India ink staining fast fixative:

[0041] 100 parts of glacial acetic acid, 200 parts of ethanol and 200 parts of methanol were ultrasonically mixed to obtain liquid A, wherein the ultrasonic power was 100 W, the ultrasonic time was 10 to 15 minutes, and the mixing temperature was room temperature;

[0042] Mix 200 parts of tert-amyl alcohol and 200 parts of formalin thoroughly to obtain solution B;

[0043] Liquid A and liquid B were ultrasonically mixed to obtain a stationary solution, the ultrasonic power was 200 W, the ultrasonic time was 20 to 25 min, and the mixing temperature was room temperature.

[0044] Example 2

[0045] 200 parts of glacial acetic acid, 400 parts of ethanol and 400 parts of methanol were ultrasonically mixed to obtain liquid A, wherein the ultrasonic power was 100 W, the ultrasonic time was 10 to 15 minutes, and the mixing temperature was room temperature;

[0046] Mix 50 parts of tert-amyl alcohol and 50 parts of formalin thoroughly to obtain solution B;

[0047] Liquid A and liquid B were ultrasonically mixed to obtain a stationary solution, the ultrasonic power was 200 W, the ultrasonic time was 20 to 25 min, and the mixing temperature was room temperature.

[0048] Example 3

[0049] 200 parts of glacial acetic acid, 300 parts of ethanol and 300 parts of methanol were ultrasonically mixed to obtain liquid A, wherein the ultrasonic power was 100 W, the ultrasonic time was 10 to 15 minutes, and the mixing temperature was room temperature;

[0050] Mix 100 parts of tert-amyl alcohol and 100 parts of formalin thoroughly to obtain solution B;

[0051] Liquid A and liquid B were ultrasonically mixed to obtain a stationary solution, the ultrasonic power was 200 W, the ultrasonic time was 20 to 25 min, and the mixing temperature was room temperature.

[0052] Example 4

[0053] 200 parts of glacial acetic acid, 400 parts of ethanol and 400 parts of methanol were ultrasonically mixed to obtain liquid A, wherein the ultrasonic power was 100 W, the ultrasonic time was 10 to 15 minutes, and the mixing temperature was room temperature;

[0054] Mix 50 parts of tert-butyl alcohol and 50 parts of formalin thoroughly to obtain solution B;

[0055] Liquid A and liquid B were ultrasonically mixed to obtain a stationary solution, the ultrasonic power was 200 W, the ultrasonic time was 20 to 25 min, and the mixing temperature was room temperature.

[0056] Experimental Example 1

[0057] The lung tissue was sampled by edge cutting to obtain a lung membrane sample, and fixed using the fixative prepared in Examples 1 to 4 above. The specific operation is as follows:

[0058] Step 1: Take a lung membrane sample from the cut edge of the lung tissue, stain it with ink, and let it stand for 30 seconds;

[0059] Step 2: Spray the fixative solution evenly on the sample stained in step 1 and let it stand for 30 seconds;

[0060] Step 3: Rinse the sample with water to wash away excess reagent after fixation.

[0061] Observe the area covered by the residual ink dye and take photos to record.

[0062] Comparative Example 1

[0063] Bouin's solution was used for fixation, and the operation steps were the same as those in Experimental Example 1.

[0064] The results are as follows:

[0065] Staining quality Example 1 Even dyeing, less residual dye, good effect Example 2 Even dyeing, less residual dye, good effect Example 3 Even dyeing, less residual dye, good effect Example 4 Uneven dyeing, less residual dye, average effect Comparative Example 1 Even dyeing, more residual dye, average effect

[0066] It can be seen from the test results in the above table that the ink staining fixative prepared by the present invention has a good staining effect and the specimens are stained evenly.

[0067] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A rapid fixing liquid for ink dyeing, characterized in that: In parts by weight, it includes the following ingredients: 100~200 parts of glacial acetic acid, 200~400 parts of ethanol, 200~400 parts of methanol, 50~200 parts of tert-amyl alcohol, 50~200 parts of formalin.

2. The fixing liquid according to claim 1, characterized in that In parts by weight, it includes the following ingredients: 200 parts of glacial acetic acid, 300 parts of ethanol, 300 parts of methanol, 100 parts of tert-amyl alcohol, Formalin 100 parts.

3. A method for preparing a fixing liquid as claimed in claim 1, characterized in that: The following steps are involved: Step 1: Evenly mix 100-200 parts of glacial acetic acid, 200-400 parts of ethanol, and 200-400 parts of methanol to obtain liquid A; Step 2: Mix 50-200 parts of tert-amyl alcohol and 50-200 parts of formalin to obtain solution B; Step 3: Ultrasonic mixing of the solution A in step 1 and the solution B in step 2 is performed to obtain a stationary solution.

4. The method for preparing a stationary liquid according to claim 3, characterized in that: The mixing method in step 1 is ultrasonic mixing, the ultrasonic power is 100W, the ultrasonic time is 10 to 15 minutes, and the mixing temperature is room temperature.

5. The method for preparing a stationary liquid according to claim 3, characterized in that: In step 3, the ultrasonic power of the ultrasonic mixing is 200 W, the ultrasonic time is 20 to 25 min, and the mixing temperature is room temperature.

6. A method for fixing pathological specimens by ink staining, using the fixing solution as claimed in claim 1 or 2, characterized in that: The following steps are involved: Step 1: Take samples from the cut edge of the pathological specimen, stain with ink, and let stand for 30 seconds; Step 2: Spray the fixative solution evenly on the sample stained in step 1 and let it stand for 30 seconds; Step 3: Rinse the sample with water to wash away excess reagent after fixation.

7. The method for fixing pathological specimens by Chinese ink staining according to claim 6, characterized in that: The size of the pathological specimen is 0.025 cm 2 ~4cm 2 .

8. A method for fixing lung membranes with India ink staining, using the fixing solution as claimed in claim 1 or 2, characterized in that: The following steps are involved: Step 1: Take a lung membrane sample from the cut edge of the lung tissue, stain it with ink, and let it stand for 30 seconds; Step 2: Spray the fixative solution evenly on the sample stained in step 1 and let it stand for 30 seconds; Step 3: Rinse the sample with water to wash away excess reagent after fixation.

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