CN107271248A - A kind of common embedding method of multigroup sample tissue - Google Patents
A kind of common embedding method of multigroup sample tissue Download PDFInfo
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- CN107271248A CN107271248A CN201710437349.5A CN201710437349A CN107271248A CN 107271248 A CN107271248 A CN 107271248A CN 201710437349 A CN201710437349 A CN 201710437349A CN 107271248 A CN107271248 A CN 107271248A
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- embedding
- sample tissue
- sample
- tissue
- animal skin
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- 238000000034 method Methods 0.000 title claims abstract description 20
- 239000012188 paraffin wax Substances 0.000 claims abstract description 20
- 241001465754 Metazoa Species 0.000 claims abstract description 19
- 230000018044 dehydration Effects 0.000 claims abstract description 5
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 5
- KDXKERNSBIXSRK-RXMQYKEDSA-N D-lysine Chemical compound NCCCC[C@@H](N)C(O)=O KDXKERNSBIXSRK-RXMQYKEDSA-N 0.000 claims abstract description 4
- 229930040373 Paraformaldehyde Natural products 0.000 claims abstract description 4
- 238000001816 cooling Methods 0.000 claims abstract description 4
- 238000001035 drying Methods 0.000 claims abstract description 4
- 229920002866 paraformaldehyde Polymers 0.000 claims abstract description 4
- 210000004706 scrotum Anatomy 0.000 claims abstract description 4
- 238000002474 experimental method Methods 0.000 claims description 6
- 230000035617 depilation Effects 0.000 claims description 2
- 230000008520 organization Effects 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 3
- 241000497192 Phyllocoptruta oleivora Species 0.000 abstract description 2
- 238000004458 analytical method Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 239000001993 wax Substances 0.000 description 4
- 238000010166 immunofluorescence Methods 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000004575 stone Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241001408627 Agriopis marginaria Species 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000000806 elastomer Substances 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000013525 hypothesis research Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000007901 in situ hybridization Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000003387 muscular Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
Classifications
-
- 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
- G01N2001/368—Mounting multiple samples in one block, e.g. TMA [Tissue Microarrays]
Abstract
Embedding method is a kind of multigroup sample tissue altogether, take the fresh and homologous buck skin of scrotum tissue of sample to be tested, it is cut into the skin bit being sized, hypodermis and fat trim is clean, it is immersed in 4 6% paraformaldehyde solutions more than 24 hours, then conventional dehydration, waxdip, fix on the supporting plate, in cooling under 0 DEG C of environment, the animal skin bit for being sized and shaped the Animal Skin of waxdip by being cut into using needs under 0 DEG C of environment after cooling, multiple pre- buried districts are vertically arranged into by above-mentioned animal skin bit by certain angle.Multiple sample tissues are separately positioned in sample tissue embedding area, then with the integral structure of paraffin embedding.Sample tissue film is adsorbed on the coated slide of poly-D-lysine, piece is baked in 60 DEG C of drying boxes 2 hours, conventional dewaxing.The present invention improves the detection and analysis precision to sample tissue, reduces the error that manual operation is caused, improves operating efficiency.
Description
Technical field
The present invention relates to a kind of paraffin body for being used to embed biological tissue, more particularly to a kind of multigroup sample tissue is wrapped altogether
Method is buried, multigroup sample biological tissue can be analyzed, observes, detects or test simultaneously, operating efficiency is improved, artificial miss is reduced
Difference.
Background technology
Paraffin embedding techniques are one of early stage tissue management techniques of paraffin section technology.Paraffin section technology existing 300
History for many years, is a technology being most widely used in histology routine techniques.It is normal that paraffin section is applied not only to observation
The morphosis of cell tissue, is also form of the subjects such as pathology and medical jurisprudence to study, observe and judge cell tissue
The main method of change, and be quite widely used in the research of many other ambits such as biology.Generally will
Pathological tissues are embedded in paraffin mass, are thinly sliced with slicer, are dyed, borrowed further according to the different demands in process of scientific research
Help pathological change, specific stain cell, tissue morphology, protein expression, Apoptosis etc. in micro- sem observation section, and then obtain
Go out to support the experimental result of scientific research hypothesis.The morphology related experiment technology of the overwhelming majority is based on paraffin section.For example：
Immunohistochemistry technology, pathological section HE staining techniques, immunofluorescence list mark technology, Double immunofluorescence technology, core in situ
Hybridization techniques, Situ Hybridization, specific stain technology（Collagenous fibres staining technique, elastomer staining technique, net
Shape stock-dye, muscular tissue staining technique, neuron（Cell）Tigroid body staining technique）Deng.
But current paraffin section, a histocyte can only be embedded per a piece of after section, so as to reduce observation, is studied
The speed and efficiency of lesion tissue.
The content of the invention
The purpose of the present invention, is to provide a kind of multigroup sample tissue embedding method altogether, embedding partition-type prepared by the present invention
Paraffin mass, multigroup sample tissue can be embedded simultaneously after section, improve the detection to sample tissue and analysis precision, reduced artificial
The error caused is operated, operating efficiency is improved.
The common embedding method of a kind of multigroup sample tissue, it is characterised in that comprise the steps：
1st, sample tissue embedding skeleton is prepared：
The fresh and homologous buck skin of scrotum tissue of sample to be tested is taken, the skin bit being sized is cut into, after depilation again
It is with scalpel that hypodermis and fat trim is clean, it is immersed in 4-6% paraformaldehyde solutions more than 24 hours, it is then conventional
Dehydration, waxdip flattens the Animal Skin after waxdip, fixes on the supporting plate, in being cooled down under 0 DEG C of environment, by waxdip after cooling
The animal skin bit that Animal Skin is sized and shaped under 0 DEG C of environment by being cut into using needs, and by embedding sample tissue quantity
Need, above-mentioned animal skin bit, which is vertically arranged by certain angle and correspondingly connected into, at least there are two sample tissues to embed area
Sample tissue embeds skeleton, standby.
2nd, tissue to be embedded is cut into multiple sample tissues being sized and shaped, multiple sample tissues is set respectively
In the sample tissue embedding area for putting the sample tissue embedding skeleton being made in step 1, while each piece of sample tissue is fixed on
In sample tissue embedding skeleton in Animal Skin adjacent thereto.Then the integral structure of paraffin embedding is used, embedding multiphase is constituted
Sample tissue paraffin mass.
Above-mentioned embedding multiphase sample tissue paraffin mass, cuts into slices along coronal-plane, in every section between sample tissue and Animal Skin
Arranged every in multiple rectangles, multiple square, multiple triangles or petal-shaped.
3rd, the interval Animal Skin in sample skeleton can stretch out paraffin in vitro, in order to recognize different embedding areas.
4th, sample tissue film is adsorbed on the coated slide of poly-D-lysine, and group position is carried out in mark
Deng mark of correlation.
5th, piece is baked in 60 DEG C of drying boxes 2 hours, conventional dewaxing carries out the subsequent operation of related experiment technology, you can.
Advantages of the present invention：
1. comparativity is higher between each group.
2. subsequent experimental reaction condition is completely the same.
3. the parameter setting of the influence experimental result such as exposure of photograph taking is completely the same.
4. same baseline is in before each group tissue detection.
5. the morphology photo of experimental result is more credible.
6. true animal skin of testing of the same race is spaced, the trouble and worry such as flake are eliminated.
Brief description of the drawings
Fig. 1 is 5mm × 12mm skin chunk schematic diagrames.
Fig. 2 is skin chunk incision site schematic diagram.
Fig. 3 is multigroup embedding device combination diagram altogether.
Fig. 4 is multigroup embedding device schematic top plan view altogether.
Fig. 5 is common embedding wax block structural upright schematic diagram.
Fig. 6 is tissue line plan in common embedding wax block.
Fig. 7, Fig. 8, Fig. 9, Figure 10 and Figure 11 are the common embedding setting planes of various configuration.
Embodiment
A kind of common embedding method of multigroup sample tissue, comprises the steps：
1 sample tissue embeds the preparation of skeleton
1.1 take same species buck scrotum portion skin histology（2cm×1cm）, and be soaked in 4% paraformaldehyde and fix 24 hours
More than.
1.2 fix the above-mentioned Animal Skin handled well, carry out conventional dehydration, fix immediately will be dynamic after waxdip, waxdip
Thing skin is flattened and is fixed on cystosepiment, is cooled down in 0 DEG C of environment.
Skin bit, is trimmed to by 1.3 finishing skin bits with scalpel：0.5cm × 1cm, rectangle（As shown in Figure 1）.
1.4 in rectangle skin bit at the broadside 5mm of side, vertical long side does the otch of 0.25cm length, and slightly extends
Kerf width, the long side of otch opposite side is changed into 7mm.（As shown in Figure 2）.
1.5 choose skin chunk according to experiment packet, and otch is relative, inserts to sample tissue bag into skin histology, is made
Bury skeleton（As shown in Figure 3,4）.
1.6 will treat investing tissue's routinely dehydration, and transparent, waxdip, according to above-mentioned cut-off shape, is repaired tissue block, will prepared
Well multiple are treated in the embedding area that investing tissue's block is separately positioned in sample tissue embedding skeleton, and are advised with 0.2mm × 15mm
The pin of lattice will treat that investing tissue's block is fixed in cut-off Animal Skin adjacent thereto, then integral with paraffin embedding
Structure.And record group position.Tissue location is as shown in Figure 5, Figure 6 in wax stone after embedding.
1.7th, cut into slices, produced along the coronal-plane of paraffin mass.Its cross sectional shape is to embed areas with four, and at four
The section of four sample tissues is embedded with embedding area.By section adsorption on the coated slide of poly-D-lysine, and in mark
The mark of correlations such as group position are carried out at note.Piece is baked in 60 DEG C of drying boxes 2 hours, conventional dewaxing carries out related experiment technology
Subsequent operation.After section, Tissue distribution is as shown in Figure 6.
1.8 is different according to experiment packet, can design the skin cut-off of various configuration, each slice plane figure such as Fig. 7, Fig. 8,
Shown in Fig. 9, Figure 10 and Figure 11.
Dotted border is wax stone border after embedding in accompanying drawing；The two longer region surrounded that separates is named as in each configuration
First quartile, arranged counterclockwise is respectively the second quadrant, third quadrant, fourth quadrant, the 5th quadrant ... so as to separator
Different groups.
Made for 4 groups of samples and the cut-off of the above, two sides of first quartile are fabricated to respectively：First quartile and end
Tail quadrant interval is set as 7mm；First quartile and the second quadrant interval are set as 6mm.After so designing, during piece is dragged for,
Without recording face anyway, order has been fixed.
Claims (2)
1. the common embedding method of a kind of multigroup sample tissue, it is characterised in that comprise the steps：
（1）, prepare sample tissue embedding skeleton：
Same kind buck scrotum skin is taken, the skin bit being sized is cut into, again with scalpel by hypodermis and fat after depilation
Fat rejects clean, is immersed in 4-6% paraformaldehyde solutions more than 24 hours, then conventional dehydration, and waxdip will be dynamic after waxdip
Thing skin is flattened, it is fixed on the supporting plate, in being cooled down under 0 DEG C of environment, after cooling by the Animal Skin of waxdip under 0 DEG C of environment by using
Need to be cut into the animal skin bit being sized and shaped, and needed by embedding sample tissue quantity, above-mentioned animal skin bit is pressed one
Determine angle and be vertically arranged and correspondingly connect into the sample tissue embedding skeleton at least with two sample tissue embedding areas, it is standby；
（2）, tissue to be embedded is cut into multiple sample tissues being sized and shaped, multiple sample tissues are set respectively
In the sample tissue embedding area for the sample tissue embedding skeleton being made in step 1, while each piece of sample tissue is fixed on into sample
In this organization embedding skeleton in cut-off Animal Skin adjacent thereto；Then the integral structure of paraffin embedding is used, embedding is constituted
Multiphase sample tissue paraffin mass；
Above-mentioned embedding multiphase sample tissue paraffin mass, cuts into slices along coronal-plane, and sample tissue and Animal Skin interval are in every section
Multiple rectangles, multiple square, multiple triangles or petal-shaped arrangement；
（3）, sample tissue film is adsorbed on the coated slide of poly-D-lysine, and carry out group position etc. in mark
Mark of correlation；
（4）, bake piece 2 hours in 60 DEG C of drying boxes, conventional dewaxing carries out the subsequent operation of related experiment technology, you can.
2. a kind of common embedding method of multigroup sample tissue according to claim 1, it is characterised in that described sample skeleton
In interval with animal skin bit stretch out paraffin it is external.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710437349.5A CN107271248B (en) | 2017-06-12 | 2017-06-12 | A kind of multiple groups sample tissue is total to embedding method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710437349.5A CN107271248B (en) | 2017-06-12 | 2017-06-12 | A kind of multiple groups sample tissue is total to embedding method |
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| Publication Number | Publication Date |
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| CN107271248A true CN107271248A (en) | 2017-10-20 |
| CN107271248B CN107271248B (en) | 2019-10-25 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201710437349.5A Active CN107271248B (en) | 2017-06-12 | 2017-06-12 | A kind of multiple groups sample tissue is total to embedding method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114383921A (en) * | 2020-10-19 | 2022-04-22 | 北京龙迈达斯科技开发有限公司 | Tissue chip and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114383921A (en) * | 2020-10-19 | 2022-04-22 | 北京龙迈达斯科技开发有限公司 | Tissue chip and preparation method thereof |
| CN114383921B (en) * | 2020-10-19 | 2023-11-21 | 北京龙迈达斯科技开发有限公司 | Tissue chip and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107271248B (en) | 2019-10-25 |
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