CN113092221A - Kit for fixing tissue slices and application thereof - Google Patents
Kit for fixing tissue slices and application thereof Download PDFInfo
- Publication number
- CN113092221A CN113092221A CN202110377940.2A CN202110377940A CN113092221A CN 113092221 A CN113092221 A CN 113092221A CN 202110377940 A CN202110377940 A CN 202110377940A CN 113092221 A CN113092221 A CN 113092221A
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- China
- Prior art keywords
- tissue
- glass slide
- quick
- staining
- drying
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- 239000011521 glass Substances 0.000 claims abstract description 33
- 238000001035 drying Methods 0.000 claims abstract description 18
- 239000003292 glue Substances 0.000 claims abstract description 17
- 235000010489 acacia gum Nutrition 0.000 claims abstract description 9
- 229930040373 Paraformaldehyde Natural products 0.000 claims abstract description 7
- 229920002866 paraformaldehyde Polymers 0.000 claims abstract description 7
- 229920000084 Gum arabic Polymers 0.000 claims abstract 5
- 241000978776 Senegalia senegal Species 0.000 claims abstract 5
- 239000000205 acacia gum Substances 0.000 claims abstract 5
- 238000000034 method Methods 0.000 claims description 13
- 230000001575 pathological effect Effects 0.000 claims description 12
- 238000010186 staining Methods 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 11
- 210000003041 ligament Anatomy 0.000 claims description 8
- 239000004819 Drying adhesive Substances 0.000 claims description 7
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 claims description 6
- 239000011248 coating agent Substances 0.000 claims description 5
- 238000000576 coating method Methods 0.000 claims description 5
- 239000001785 acacia senegal l. willd gum Substances 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 238000005520 cutting process Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 238000011532 immunohistochemical staining Methods 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- 238000003892 spreading Methods 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 239000006101 laboratory sample Substances 0.000 claims 1
- 230000007170 pathology Effects 0.000 claims 1
- 238000012800 visualization Methods 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 238000004043 dyeing Methods 0.000 abstract description 9
- 238000003364 immunohistochemistry Methods 0.000 abstract description 2
- 210000001519 tissue Anatomy 0.000 description 35
- 108010010803 Gelatin Proteins 0.000 description 4
- 229920000159 gelatin Polymers 0.000 description 4
- 239000008273 gelatin Substances 0.000 description 4
- 235000019322 gelatine Nutrition 0.000 description 4
- 235000011852 gelatine desserts Nutrition 0.000 description 4
- 229910052720 vanadium Inorganic materials 0.000 description 3
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- LEONUFNNVUYDNQ-UHFFFAOYSA-N vanadium atom Chemical compound [V] LEONUFNNVUYDNQ-UHFFFAOYSA-N 0.000 description 2
- 244000017020 Ipomoea batatas Species 0.000 description 1
- 235000002678 Ipomoea batatas Nutrition 0.000 description 1
- 241000406668 Loxodonta cyclotis Species 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- HBXWYZMULLEJSG-UHFFFAOYSA-N chromium vanadium Chemical compound [V][Cr][V][Cr] HBXWYZMULLEJSG-UHFFFAOYSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
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- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a kit for fixing tissue slices and application thereof; the kit comprises a glass slide, paraformaldehyde, a gum arabic solution and quick-drying glue. The quick-drying glue is dripped on the glass slide and evenly smeared into a thin layer, the tissue section is quickly adhered on the glass slide on which the quick-drying glue is hung, the tissue section is tightly combined with the glass slide, the phenomenon of slide falling is hardly caused, the operation is simple, the used glass slide has lower price than the glass slide with glue on the market, and the quick-drying glue can be used for various dyeing such as HE dyeing, immunohistochemistry and the like.
Description
This application is a divisional application entitled "method for fixing tissue slices and use thereof" filed on 2015, 07, 16, and having application number of 201510419068.8.
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a kit for fixing a tissue slice and application thereof.
Background
Tissue section shedding is always an inevitable problem in the pathological staining process. The existing method for preventing the slice from falling mainly comprises collodion coating, polylysine and chromium-vanadium gelatin treatment slide and the like. Although hard tissue sections can be prevented from falling, the sections are too thick (about 40 to 100 μm), and therefore, overlapping of tissues is not easy to observe, and pathological staining cannot be performed. Paraffin and frozen sections can reach within 10 mu m, cell morphology can be observed well, but the problems that some tissue patches are not firm all the time and the slices are easy to drop during dyeing exist, and especially in the section dyeing of tissue engineering materials such as silk, artificial ligaments and the like, the slice dropping phenomenon is more prominent. Meanwhile, in the dyeing of three colors of HE and ponceau, immunohistochemistry and the like, the tissue slide dropping phenomenon is particularly serious due to the complex dyeing steps, the overlong tissue soaking time and the like.
Disclosure of Invention
The invention aims to provide a tissue slice fixing method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme.
A method of tissue slice fixation comprising the steps of:
1) fixing experimental materials or surgical pathological specimens by using paraformaldehyde, and performing decalcification treatment after fixing the experimental materials or the surgical pathological specimens by using the paraformaldehyde if the experimental materials or the surgical pathological specimens are bone tissues;
2) soaking the experimental material or the surgical pathological specimen treated in the step 1) in an Arabic gum solution for 24-48 hours, and then cutting the specimen to 5-10 mu m by a freezing microtome to obtain a tissue section;
3) and (3) uniformly coating the quick-drying adhesive on the glass slide, fixing the tissue slice on the region uniformly coated with the quick-drying adhesive on the glass slide, and naturally drying or baking the tissue slice for 1-2 hours at 55-65 ℃ by using a baking sheet machine.
The tissue section is obtained from all experimental materials or surgical pathological specimens which are easy to fall.
The experimental material or the surgical pathological specimen is silk tissue engineering ligament.
The method for uniformly coating the quick-drying adhesive on the glass slide specifically comprises the following steps: dripping a plurality of drops of quick-drying glue on the glass slide A, immediately covering the upper surface of the glass slide A with the glass slide B, and horizontally spreading the quick-drying glue dripped on the glass slide A, so that the quick-drying glue is uniformly coated on the glass slide A and the glass slide B.
The application of the fixing method of the tissue section in tissue staining.
The tissue sections were used for HE staining, ponceau trichrome staining or immunohistochemical staining.
The tissue section is obtained from a silk tissue engineering ligament.
The invention has the beneficial effects that:
after a thin layer of quick-drying adhesive is uniformly coated on a common glass slide, the frozen tissue section is pasted, the tissue section is tightly combined with the glass slide, the preparation is convenient, the operation is simple, the processing time is short, the cost is low, the sheet dropping phenomenon in the dyeing process is effectively avoided, the sheet dropping problem of pathological dyeing is thoroughly solved, the quick-drying tissue section is particularly suitable for the section dyeing of easily dropped silk materials, tissue engineering ligaments or other tissue engineering materials, and the problems that hard tissue sections are too thick and can not clearly display structures such as cells are effectively solved.
Drawings
FIG. 1 shows the results of fixing silk ligament sections on a glass slide treated with chrome-vanadium gelatin and HE staining;
FIG. 2 shows the results of fixing silk ligament sections on quick-drying glue-coated glass slides and HE staining.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and examples.
Taking silk ligament tissue fixed and decalcified by paraformaldehyde, soaking for 24-48 hours by using an Arabic gum solution (an aqueous solution containing 10% of Arabic gum and 30% of sucrose by mass fraction) to obtain a sample, cutting the sample into tissue slices with the thickness of 5-10 microns by using a freezing microtome, dripping 1 drop of quick-drying glue (such as elephant card glue in green cleaning daily necessities of Ipomoea batatas) on a common glass slide, uniformly coating the quick-drying glue on two glass slides by using the quick tabletting method of the other glass slide, and quickly adhering the tissue slices to the area coated with the quick-drying glue on one glass slide; then the slices are naturally dried or baked on a baking machine (60 ℃) for 1-2 hours. Control groups used slides treated with chrome vanadium gelatin to adhere to the tissue sections while HE staining was performed.
Referring to fig. 1 and 2, severe tissue loss after staining can be seen in fig. 1 (chrome vanadium gelatin treated slides); in FIG. 2, the staining color was normal (the slide treated by the method of the present invention), and there was almost no chipping.
The tissue slice fixing method can effectively avoid the phenomenon of slice dropping (slice dropping) in the tissue staining process.
Claims (8)
1. A kit for fixing tissue sections, comprising a glass slide, paraformaldehyde, a gum arabic solution, and quick-drying glue.
2. The kit according to claim 1, wherein the gum arabic solution is an aqueous gum arabic solution comprising, in mass fraction, 10% gum arabic and 30% sucrose.
3. Use of a kit according to claim 1 or 2 for the fixation of tissue sections, wherein said tissue sections are obtained from all experimental or surgical pathological specimens susceptible to flaking.
4. Use according to claim 3, wherein the method of fixing tissue slices comprises the following steps:
1) fixing the experimental material or the surgical pathological specimen with the paraformaldehyde;
2) soaking the experimental material or the surgical pathological specimen treated in the step 1) in the water solution of the arabic gum for 24-48 hours, and then cutting the specimen to 5-10 mu m by a freezing microtome to obtain a tissue section;
3) uniformly coating the quick-drying adhesive on a glass slide, fixing the tissue slice on the area uniformly coated with the quick-drying adhesive on the glass slide, and naturally drying the tissue slice or baking the tissue slice for 1-2 hours at 55-65 ℃ by using a baking sheet machine;
5. use according to claim 4, characterized in that it comprises, when said laboratory sample or surgical pathology specimen is a silk tissue engineering ligament, a step of decalcification after paraformaldehyde fixation.
6. The use according to claim 4, wherein the uniform application of the quick-drying adhesive to the glass slide comprises the following steps: dripping a plurality of drops of quick-drying glue on the glass slide A, immediately covering the upper surface of the glass slide A with the glass slide B, and horizontally spreading the quick-drying glue dripped on the glass slide A, so that the quick-drying glue is uniformly coated on the glass slide A and the glass slide B.
7. Use of a kit according to claim 1 or 2 for avoiding slide-off in tissue staining, wherein the tissue staining is HE staining, ponceau trichrome staining or immunohistochemical staining.
8. Use of the kit according to claim 1 or 2 for increasing the clarity of cell visualization in hard tissue stained sections, wherein the tissue staining is HE staining, ponceau trichrome staining or immunohistochemical staining.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110377940.2A CN113092221A (en) | 2015-07-16 | 2015-07-16 | Kit for fixing tissue slices and application thereof |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510419068.8A CN105004594A (en) | 2015-07-16 | 2015-07-16 | Fixing method and use of tissue slice |
| CN202110377940.2A CN113092221A (en) | 2015-07-16 | 2015-07-16 | Kit for fixing tissue slices and application thereof |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510419068.8A Division CN105004594A (en) | 2015-07-16 | 2015-07-16 | Fixing method and use of tissue slice |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113092221A true CN113092221A (en) | 2021-07-09 |
Family
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Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110377940.2A Pending CN113092221A (en) | 2015-07-16 | 2015-07-16 | Kit for fixing tissue slices and application thereof |
| CN201510419068.8A Pending CN105004594A (en) | 2015-07-16 | 2015-07-16 | Fixing method and use of tissue slice |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510419068.8A Pending CN105004594A (en) | 2015-07-16 | 2015-07-16 | Fixing method and use of tissue slice |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN113092221A (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108651438B (en) * | 2018-05-02 | 2019-04-05 | 济南天灿医疗科技有限公司 | A kind of fixed agents for defoliating of tissue for tissue block and application |
| CN111238901B (en) * | 2020-03-04 | 2023-03-03 | 四川省人民医院 | Manufacturing method of improved mouse eyeball frozen section |
| CN112985952B (en) * | 2021-02-20 | 2022-09-20 | 山东骏腾医疗科技有限公司 | Pathological sealing sheet material |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100047860A1 (en) * | 2007-03-29 | 2010-02-25 | National University Corporation University of Toya ma | Device for storing specimen slice and instrument for microscopic observation provided with the same |
| CN104062165A (en) * | 2014-06-17 | 2014-09-24 | 塔里木大学 | Anti-dropping agent for slices, and preparation method and usage of anti-dropping agent |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103471903B (en) * | 2013-09-29 | 2016-01-06 | 吴天骥 | The collocation method of section immobile liquid and application thereof |
| CN104634626A (en) * | 2013-11-15 | 2015-05-20 | 上海交通大学 | Improved frozen slicing method and applications thereof |
| CN103884560A (en) * | 2014-04-10 | 2014-06-25 | 中国水产科学研究院长江水产研究所 | Preparation method and application of Chinese giant salamander phalanx section |
| CN104215471A (en) * | 2014-09-15 | 2014-12-17 | 东华大学 | Preparation method of microRNA in-situ hybridized frozen section |
-
2015
- 2015-07-16 CN CN202110377940.2A patent/CN113092221A/en active Pending
- 2015-07-16 CN CN201510419068.8A patent/CN105004594A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100047860A1 (en) * | 2007-03-29 | 2010-02-25 | National University Corporation University of Toya ma | Device for storing specimen slice and instrument for microscopic observation provided with the same |
| CN104062165A (en) * | 2014-06-17 | 2014-09-24 | 塔里木大学 | Anti-dropping agent for slices, and preparation method and usage of anti-dropping agent |
Non-Patent Citations (1)
| Title |
|---|
| 滕可导 主编: "《彩图家畜组织学与胚胎学实验指导》", vol. 1, 中国农业大学出版社, article 操石磊, pages: 154 - 179 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105004594A (en) | 2015-10-28 |
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| PB01 | Publication | ||
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| RJ01 | Rejection of invention patent application after publication | ||
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Application publication date: 20210709 |