CN114018680B - Manufacturing method of choroid melanoma specimen wax block and chip - Google Patents
Manufacturing method of choroid melanoma specimen wax block and chip Download PDFInfo
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- CN114018680B CN114018680B CN202111267120.4A CN202111267120A CN114018680B CN 114018680 B CN114018680 B CN 114018680B CN 202111267120 A CN202111267120 A CN 202111267120A CN 114018680 B CN114018680 B CN 114018680B
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- 206010008773 Choroid melanoma Diseases 0.000 title claims abstract description 12
- 201000002742 malignant choroid melanoma Diseases 0.000 title claims abstract description 12
- 238000004519 manufacturing process Methods 0.000 title claims description 6
- 239000001993 wax Substances 0.000 claims abstract description 56
- 239000012188 paraffin wax Substances 0.000 claims abstract description 43
- 229920000742 Cotton Polymers 0.000 claims abstract description 31
- 239000007788 liquid Substances 0.000 claims abstract description 23
- 238000002844 melting Methods 0.000 claims abstract description 21
- 230000008018 melting Effects 0.000 claims abstract description 21
- 208000005156 Dehydration Diseases 0.000 claims abstract description 12
- 230000018044 dehydration Effects 0.000 claims abstract description 12
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000011358 absorbing material Substances 0.000 claims abstract description 5
- 238000001816 cooling Methods 0.000 claims abstract description 4
- 201000001441 melanoma Diseases 0.000 claims description 15
- 206010028980 Neoplasm Diseases 0.000 claims description 5
- 239000010902 straw Substances 0.000 claims description 5
- 238000005520 cutting process Methods 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 2
- 239000004745 nonwoven fabric Substances 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 abstract description 3
- 229940057995 liquid paraffin Drugs 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 239000012189 cellwax Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 206010003445 Ascites Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000009928 pasteurization Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 210000004910 pleural fluid Anatomy 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
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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
- G01N1/36—Embedding or analogous mounting of samples
-
- 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
- G01N1/36—Embedding or analogous mounting of samples
- G01N2001/366—Moulds; Demoulding
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention discloses a preparation method of a choroid melanoma specimen wax block, which does not adopt conventional centrifugation, but adopts a standing method to naturally sink powdered tissues, uses a suction tube to absorb the precipitated tissues, then slowly drops the absorbed tissues containing liquid on cotton yarn paper filled with a water absorbing material at the bottom to absorb water, then wraps the tissues in the cotton yarn paper for dehydration treatment, then uses paraffin with a lower melting point to embed, takes out the tissues after the liquid paraffin is solidified, and removes the cotton yarn paper. The preparation method comprises the steps of transferring the wax block into a groove of a wax mold made of paraffin with a higher melting point on the basis of the choroid melanoma specimen wax block, controlling the temperature to enable the paraffin to be fused, naturally sinking choroid melanoma glass-cut tissues, cooling and solidifying to prepare the choroid melanoma specimen chip.
Description
Technical Field
The invention relates to a preparation method of a choroid melanoma specimen wax block and a tissue chip thereof.
Background
Pathological tissues taken from a patient through operation or puncture are sent to a pathology department for treatment, and finally, the tissues are wrapped in paraffin and stored in the form of a wax block, wherein the wax block is called a tissue wax block which can be further prepared into pathological sections for clinical diagnosis, so the tissue wax block is important clinical data.
The tissues obtained from some liquid specimens (such as pleural fluid, ascites, etc.) are very fine and fragmented, and making wax masses from these fragmented tissues and even cells is much more cumbersome than making large wax masses of tissues. The cell paraffin block is prepared by extracting cells in liquid, fixing with paraffin, and making into tissue section-like form. The conventional preparation steps of the cell wax block are that supernatant liquid is removed from extracted body fluid in a centrifugal mode, coagulated precipitates are collected after centrifugation to prepare the cell wax block, most body fluid specimens contain red blood cells and have certain cohesiveness, and the red blood cells are not easy to disperse after being prepared into the wax block and can be prepared into a cell chip conventionally, and the excised tissues of the vitreous humour of the choroidal melanoma are both melanoma cells and have no viscous components such as red blood cells and the like and have no cohesive force.
Therefore, those skilled in the art are devoted to develop a simple and easy method for preparing the wax block of the excised tissue of the vitreous tumor of the choroidal melanoma and the tissue chip thereof.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a method for preparing a choriomelanoma vitrectomy tissue wax block, which comprises the following steps:
1) Standing the vitrified liquid mixed with the excised tissue of the vitreous humour of the choroid melanoma to naturally sink the powdered tissue;
2) Inserting a suction tube into the bottom to suck the precipitated powdered tissue;
3) Slowly dripping the sucked tissue containing liquid on cotton yarn paper filled with a water-absorbing material, and naturally filtering the cotton yarn paper to obtain a powdery tissue;
4) Wrapping the powdered tissue absorbing the water in cotton yarn paper for dehydration treatment;
5) Wrapping the powdery tissue in cotton yarn paper after dehydration, and immersing the cotton yarn paper and the tissue in liquid first paraffin;
6) And cutting out a wax block containing a powdery tissue after the liquid first paraffin is cooled and solidified, and removing the cotton yarn paper.
Preferably, the straw in step 2) is a plastic pasteurisation straw.
Preferably, the water absorbing material in step 3) is one or more of paper towel, non-woven fabric, filter paper and straw paper. In one embodiment of the invention, the cotton gauze paper is laid on the tissue paper in advance, then the sucked powdery tissue containing liquid is slowly dripped on the cotton gauze paper, the tissue paper on the back of the cotton gauze paper directly sucks the liquid to retain the tissue on the cotton gauze paper, and then the tissue is wrapped by the cotton gauze paper for dehydration treatment.
Further, the dehydration treatment in the step 4) refers to a conventional dehydration treatment manner, such as conventional dehydration in a dehydrator.
Preferably, the melting point of the first paraffin wax is less than 60 ℃, preferably between 55 and 58 ℃.
Preferably, the temperature of the liquid first paraffin wax in step 5) is 1 to 3 ℃ above the melting point.
Furthermore, the invention provides a preparation method of the choroidal melanoma specimen chip, which comprises the following steps after the preparation method of the choroidal melanoma specimen wax block:
a) Transferring the choriomelanoma specimen wax block into a recess of a wax mold made of a second paraffin wax, wherein the melting point of the second paraffin wax is higher than that of the first paraffin wax;
b) Placing the wax mould in an environment with the temperature higher than the melting point of the first paraffin and lower than the melting point of the second paraffin, fusing the first paraffin with the wax mould after melting, and naturally sinking the choroid melanoma specimen;
c) Cooling and solidifying.
Preferably, the second paraffin wax has a melting point of greater than 60 ℃, preferably 75 to 80 ℃.
Preferably, the temperature in step b) is 60 ± 1 ℃.
In a preferred embodiment of the present invention, the step a) transfers the wax block made of the first paraffin wax into the recess of the wax pattern made of the second paraffin wax by cutting the desired wax block with a blade and then transferring it into the recess with a nipper.
Preferably, the groove in step a) is a tapered groove with a small bottom. The middle of the conventional tissue chip wax mould is provided with a through hole, so that the excised tissue of the powdery choroid melanoma vitreous lump which is not easy to agglomerate easily leaks out from the bottom.
Preferably, the number of the grooves in the step a) is plural; in a preferred embodiment of the invention, the grooves are arranged in a 6 x 4 array; in another preferred embodiment of the invention, the grooves are arranged in a 9 x 10 array.
Furthermore, the invention provides a wax mould for manufacturing the choroidal melanoma specimen chip, wherein one surface of the wax mould is a smooth plane, and a plurality of conical grooves with small bottom openings are arranged on the other corresponding surface of the wax mould.
Further, the present invention provides a choriomelanoma specimen wax block and a specimen chip manufactured by the above method.
The invention relates to a scheme specially customized for a powdery choroidal melanoma vitreous tumor excision tissue which is not easy to coagulate, which does not adopt a conventional centrifugation, fixation and substrate taking method, but naturally deposits the powdery tissue after standing, sucks the deposited tissue by a suction pipe, slowly drops the sucked tissue containing liquid on cotton gauze paper with a water absorption material below to absorb water, wraps the tissue in the cotton gauze paper to carry out dehydration treatment, then embeds the tissue by paraffin with a lower melting point, takes out the tissue after the liquid paraffin is solidified, removes the cotton gauze paper, and has simple and easy operation steps. In the prior art, the powdery tissues leak from the hollow wax mould, the conical groove wax mould provided by the invention can enable the tissues to be gathered at the bottom of the groove but not leak, and the wax layer at the conical bottom is adopted for slicing, so that the tissues at each point position can be ensured.
The conception, the specific structure and the technical effects of the present invention will be further described with reference to the accompanying drawings to fully understand the objects, the features and the effects of the present invention.
Drawings
FIG. 1 is a schematic top surface structure of a preferred embodiment of a wax pattern of the present invention;
fig. 2 is a side view schematically illustrating a wax pattern according to a preferred embodiment of the present invention.
Detailed Description
EXAMPLE 1 preparation of sample wax masses
1) Injecting the vitrified liquid mixed with the excised tissue of the vitreous tumor of the choroidal melanoma into a centrifuge tube, standing for 10 minutes, and naturally sinking the powdered tissue;
2) Sucking the substrate mixed with the powdered tissue with a plastic pasteur pipette;
3) The square cotton yarn paper to be used for covering the tissue is laid on the paper towel, liquid in the Pasteur straw is slowly dripped on the cotton yarn paper, and irrelevant liquid is absorbed by the paper towel on the back of the cotton yarn paper, so that the powdery tissue is retained on the cotton yarn paper;
4) Wrapping the powdery tissue with cotton gauze paper, and placing the cotton gauze paper into a dehydrator for dehydration;
5) After dehydration, wrapping the tissue in cotton yarn paper, and immersing the tissue in liquid first paraffin with the melting point of 58 ℃, wherein the temperature of the liquid first paraffin can be about 60 ℃, so that the tissue is fully soaked in the paraffin;
6) Cooling to solidify the first paraffin;
7) Cutting the first paraffin block to obtain a tissue wax block, and removing cotton gauze paper to obtain the choroidal melanoma specimen wax block.
EXAMPLE 2 preparation of sample chip
Referring to fig. 1 and 2, a rectangular wax pattern for manufacturing a sample chip according to the present invention is shown, wherein the lower surface of the wax pattern is a smooth plane 1, and the upper surface 2 of the wax pattern is densely arranged with 9 × 10 conical grooves 3.
The preparation steps of the sample chip in the embodiment are as follows:
transferring the obtained choriomelanoma specimen wax block into a groove 3 of a wax mould made of second paraffin, wherein the melting point of the second paraffin is 80 ℃; if the first paraffin is not solidified, the tissue can also be aspirated with a heated glass pipette.
And (3) placing the whole wax mold in a thermostat at 60 ℃ for a period of time, so that the first paraffin in the groove is melted and then is fused with the wax mold into a whole. The wax mould cannot be melted but can be softened due to the high melting point and slightly fused with paraffin in the conical groove, the paraffin in the groove is in a liquid state, the tissue of the choroid melanoma vitreous tumor after excision can naturally sink, then the wax mould is taken out to room temperature, and the specimen chip is prepared after the bottom of the groove is solidified.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations can be devised by those skilled in the art in light of the above teachings. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.
Claims (10)
1. A preparation method of a wax block for a choroidal melanoma specimen is characterized by comprising the following steps:
1) Standing the vitrified liquid mixed with the excised tissue of the vitreous tumor of the choroidal melanoma to naturally sink the powdered tissue;
2) Inserting a suction tube into the bottom to suck the precipitated powdered tissue;
3) Slowly dripping the sucked tissue containing liquid on cotton yarn paper filled with a water-absorbing material, and naturally filtering the cotton yarn paper to obtain a powdery tissue;
4) Wrapping the powdered tissue absorbing the water in cotton yarn paper for dehydration treatment;
5) Wrapping the powdery tissue in cotton yarn paper after dehydration, and immersing the cotton yarn paper and the tissue in liquid first paraffin;
6) And cutting out the wax block containing the powdery tissue after the liquid first paraffin is cooled and solidified, and removing the cotton yarn paper.
2. The method for preparing the choriomelanoma specimen wax block according to claim 1, wherein the water-absorbing material is one or more of paper towel, non-woven fabric, filter paper and straw paper.
3. The method of claim 1, wherein the first paraffin wax has a melting point of less than 60 ℃.
4. The method for preparing the choroidal melanoma specimen wax block according to claim 1, wherein the temperature of the liquid first paraffin in the step 5) is 1-3 ℃ higher than the melting point.
5. A manufacturing method of a choroid melanoma specimen chip is characterized by comprising the following steps:
a) Transferring a choriomelanoma sample wax block prepared according to any one of claims 1 to 4 into a recess of a wax mold made of a second paraffin wax, wherein the second paraffin wax has a melting point higher than the melting point of the first paraffin wax;
b) Placing the wax mould in an environment with the temperature higher than the melting point of the first paraffin and lower than the melting point of the second paraffin, melting the first paraffin, fusing the first paraffin with the wax mould, and naturally sinking the choroid melanoma specimen;
c) Cooling and solidifying.
6. The method of claim 5, wherein the second paraffin wax has a melting point of greater than 60 ℃.
7. The method for preparing a choroidal melanoma specimen chip according to claim 5, wherein the temperature in step b) is 60 ± 1 ℃.
8. The method for preparing a choroidal melanoma specimen chip according to claim 5, wherein the groove in step a) is a tapered groove with a small bottom.
9. A choriomelanoma sample wax block produced by the method of producing a choriomelanoma sample wax block according to any one of claims 1 to 4.
10. A choriomelanoma sample chip produced by the method for producing a choriomelanoma sample chip according to any one of claims 5 to 8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111267120.4A CN114018680B (en) | 2021-10-28 | 2021-10-28 | Manufacturing method of choroid melanoma specimen wax block and chip |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111267120.4A CN114018680B (en) | 2021-10-28 | 2021-10-28 | Manufacturing method of choroid melanoma specimen wax block and chip |
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| CN114018680A CN114018680A (en) | 2022-02-08 |
| CN114018680B true CN114018680B (en) | 2022-10-21 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FI83668C (en) * | 1989-09-08 | 1991-08-12 | Johan Eric Walter Ahlqvist | Procedures and agents for the treatment of cytological preparations |
| CN1204405C (en) * | 2001-09-13 | 2005-06-01 | 陕西超英生物医学研究开发有限公司 | Tissue microarray biochip |
| AU2002352787A1 (en) * | 2001-11-20 | 2003-06-10 | Genentech, Inc. | Cell and tissue arrays and microarrays and methods of use |
| CN104122132A (en) * | 2014-07-31 | 2014-10-29 | 北京海思特临床检验所有限公司 | Hydrothorax and ascite malignant cell embedding method |
| CN106769174B (en) * | 2016-11-22 | 2018-07-13 | 南京林业大学 | Plant tissue slice receptor blank wax stone boring instrument |
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