CN108827747A - A kind of plasticity embedding method of multicolor fluorescence marker samples - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 32
- 239000003550 marker Substances 0.000 title claims abstract description 14
- 239000011347 resin Substances 0.000 claims abstract description 44
- 229920005989 resin Polymers 0.000 claims abstract description 44
- 239000012472 biological sample Substances 0.000 claims abstract description 26
- 239000007788 liquid Substances 0.000 claims abstract description 23
- 230000035515 penetration Effects 0.000 claims abstract description 20
- 239000000523 sample Substances 0.000 claims abstract description 17
- 230000008595 infiltration Effects 0.000 claims abstract description 9
- 238000001764 infiltration Methods 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 24
- 229930040373 Paraformaldehyde Natural products 0.000 claims description 12
- 239000005457 ice water Substances 0.000 claims description 12
- 229920002866 paraformaldehyde Polymers 0.000 claims description 12
- 235000019441 ethanol Nutrition 0.000 claims description 10
- 230000018044 dehydration Effects 0.000 claims description 8
- 238000006297 dehydration reaction Methods 0.000 claims description 8
- 239000002904 solvent Substances 0.000 claims description 5
- 230000001476 alcoholic effect Effects 0.000 claims description 4
- 241001465754 Metazoa Species 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 2
- VOZRXNHHFUQHIL-UHFFFAOYSA-N glycidyl methacrylate Chemical compound CC(=C)C(=O)OCC1CO1 VOZRXNHHFUQHIL-UHFFFAOYSA-N 0.000 description 27
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 21
- 210000001519 tissue Anatomy 0.000 description 18
- 210000005013 brain tissue Anatomy 0.000 description 15
- 238000006116 polymerization reaction Methods 0.000 description 15
- 238000002360 preparation method Methods 0.000 description 11
- 210000005003 heart tissue Anatomy 0.000 description 9
- 238000001816 cooling Methods 0.000 description 7
- 238000003384 imaging method Methods 0.000 description 7
- SOGAXMICEFXMKE-UHFFFAOYSA-N Butylmethacrylate Chemical compound CCCCOC(=O)C(C)=C SOGAXMICEFXMKE-UHFFFAOYSA-N 0.000 description 6
- 108010054624 red fluorescent protein Proteins 0.000 description 5
- 229930006000 Sucrose Natural products 0.000 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 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 239000005090 green fluorescent protein Substances 0.000 description 3
- 239000000178 monomer Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 238000001291 vacuum drying Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 108091006047 fluorescent proteins Proteins 0.000 description 2
- 102000034287 fluorescent proteins Human genes 0.000 description 2
- MURGITYSBWUQTI-UHFFFAOYSA-N fluorescin Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC=C(O)C=C2OC2=CC(O)=CC=C21 MURGITYSBWUQTI-UHFFFAOYSA-N 0.000 description 2
- 230000010412 perfusion Effects 0.000 description 2
- 238000010377 protein imaging Methods 0.000 description 2
- OALHHIHQOFIMEF-UHFFFAOYSA-N 3',6'-dihydroxy-2',4',5',7'-tetraiodo-3h-spiro[2-benzofuran-1,9'-xanthene]-3-one Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 OALHHIHQOFIMEF-UHFFFAOYSA-N 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 1
- 108010047357 Luminescent Proteins Proteins 0.000 description 1
- 102000006830 Luminescent Proteins Human genes 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000256856 Vespidae Species 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 108010021843 fluorescent protein 583 Proteins 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
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- 239000007787 solid Substances 0.000 description 1
- YCUVUDODLRLVIC-VPHDGDOJSA-N sudan black b Chemical compound C1=CC(=C23)NC(C)(C)NC2=CC=CC3=C1\N=N\C(C1=CC=CC=C11)=CC=C1\N=N\C1=CC=CC=C1 YCUVUDODLRLVIC-VPHDGDOJSA-N 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
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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
- G01N1/30—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
- G01N2001/364—Embedding or analogous mounting of samples using resins, epoxy
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- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
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- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
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Abstract
The present invention discloses a kind of multicolor fluorescence marker samples plasticity embedding method, including：Biological sample is pre-processed；Pretreated biological sample is sequentially placed into the resin solution of the SBB and DTT of different quality concentration and carries out gradient penetration, each concentration 0.5-3 hours, is then placed again into the resin solution of fresh addition SBB and DTT and continue infiltration 6-12 hours；The fluorescent marker sample that is embedded after being embedded, being heated to the biological sample after gradient penetration with resin penetration liquid.Embedding method provided by the invention is embedded suitable for various kinds of resin, while can be suitably used for multiple color fluorescent marker sample.
Description
Technical field
The present invention relates to a kind of technical field of bioengineering, and in particular to a kind of multicolor fluorescence marker samples plasticity embedding side
Method.
Background technique
Currently, by fluorescent labelling techniques various information can be marked simultaneously on a sample, to further investigation biology
The structure and function of body plays a significant role.
Existing resin embedding method embeds biological group using pure resin or the resin sample infiltration for dissolving SBB
It knits, can get the sample of background fluorescence reduction, it is disadvantageous in that：This embedding method is only applicable to green fluorescent protein
The biological tissue of label is not suitable for other fluorescence (such as red fluorescent protein, various fluoresceins probes etc.) label, thus
Its application is greatly limited, such as shown in Fig. 1.
Summary of the invention
In view of this, the present invention provides a kind of embedding method that can be suitably used for multiple color fluorescent marker sample.
In order to solve the above technical problems, the technical scheme is that being embedded using a kind of multicolor fluorescence marker samples plasticity
Method, including：
Biological sample is pre-processed；
Pretreated biological sample is sequentially placed into the alcoholic solution of the gradient concentration of 0.5-0.7wt%DTT and is carried out
Infiltration, each concentration 0.5-3 hour, then continue in the resin solution of different quality concentration permeate 6-12 hours, it is described not
Resin solution with concentration includes the DTT of the SBB and 0.7-1wt% of 0.1-0.2wt%；
The fluorescence that is embedded after being embedded, being heated to the biological sample after gradient penetration with resin penetration liquid
Marker samples.
Preferably, it is described to biological sample carry out pretreatment include：
Biological sample is fixed 12-24 hours in paraformaldehyde fixer；
Biological sample after fixation is rinsed 6-12 hours in rinsing liquid；
Biological sample after the rinsing is sequentially placed into 50%, 70% and 95% or 100% ethyl alcohol of ice water precooling
Serial dehydration is carried out in solution, 5 minutes to 2 hours every time.
Preferably, the mass concentration of the resin solution of the various concentration be 40%~60%, 60%~70%, 70~
90% and 90%~100%.
Preferably, the mass concentration of the resin solution of the various concentration is respectively 50%, 70%, 85% and 100%.
Preferably, the mass concentration of the gradient concentration alcoholic solution is respectively 50%, 70%, 85% and 100%.
Preferably, described that the biological sample after gradient penetration is embedded specially with resin penetration liquid：
Biological sample after gradient penetration is placed in imbedded mold, is then full of resin penetration liquid in a mold.
Preferably, the resin in the resin penetration liquid and resin solution is specially GMA resin.
Preferably, the fluorescent marker sample embedded after heating is specially：
The filled mold is heated 12 to 24 hours at 35 DEG C -40 DEG C.
Preferably, the solvent of the resin solution is 95% ethyl alcohol.
Preferably, the biological sample is selected from the animal tissue that can emit fluorescence.
Primary improvements of the invention are to provide a kind of plasticity embedding method of multicolor fluorescence marker samples：This method
It is permeated using DTT, and animal tissue can be made to keep original fluorescence signal completely using lower polymerization temperature, together
When carry out infiltration and lower polymerization temperature using SBB solution and can significantly reduce the background fluorescence of resin embedding biological sample,
And process of osmosis is relatively simple, can be suitable for the biological sample of various sizes；And due to first having carried out the infiltration of gradient, then
It is embedded so that sample is able to satisfy continuous slice with certain degree of hardness；It is able to maintain the antigen active of biological tissue；Gained
The biological sample of embedding can apply to the imaging of block face and combine the technical of continuous slice, can be used for quick obtaining fluorescin mark
The high-resolution three-dimensional fluorescence distribution for remembering tissue, then carries out dependency structure and functional study.
Detailed description of the invention
Fig. 1 is the brain tissue tdTomato red fluorescent protein imaging results of conventional resins embedding method preparation；
Fig. 2 is that the brain tissue tdTomato of the resin embedding method preparation for the high s/n ratio that the embodiment of the present invention 1 embeds is red
Color imaging fluorescent protein result；
Fig. 3 is that the brain tissue mCherry of the resin embedding method preparation for the high s/n ratio that the embodiment of the present invention 1 embeds is red
Imaging fluorescent protein result；
Fig. 4 is that the brain tissue DsRed red of the resin embedding method preparation for the high s/n ratio that the embodiment of the present invention 1 embeds is glimmering
Photoprotein imaging results；
Fig. 5 is the brain tissue green fluorescence egg of the resin embedding method preparation for the high s/n ratio that the embodiment of the present invention 1 embeds
White (GFP) imaging results；
Fig. 6 is the brain tissue blue-fluorescence egg of the resin embedding method preparation for the high s/n ratio that the embodiment of the present invention 1 embeds
White (BFP) imaging results；
Fig. 7 is in the heart tissue of resin embedding method preparation for the high s/n ratio that the embodiment of the present invention 2 embeds
TdTomato red fluorescent protein imaging results；
Fig. 8 is red glimmering in the myeloid tissue of the resin embedding method preparation for the high s/n ratio that the embodiment of the present invention 3 embeds
Light element imaging results.
Specific embodiment
In order to make those skilled in the art more fully understand technical solution of the present invention, With reference to embodiment
The present invention is described in further detail.
Explanation of nouns：
GMA resin：Glycidyl methacrylate.
DTT：Dithiothreitol (DTT).
SBB：Sudan black B
Technology incidence of criminal offenses of the invention is further described below by specific embodiment.
Embodiment 1
To adult VIP::Ai14 is hybridized mouse and is anaesthetized using 1% yellow Jackets, and the mode of heart perfusion is taken to be filled
Stream, is first perfused 0.01M PBS solution 3-7 minutes of 37 DEG C, and 4% paraformaldehyde fixer 7-10 points of ice water pre-cooling are perfused later
Clock.After heart perfusion, Mice brain tissues are taken, are put into 4% paraformaldehyde fixer, the rear fixed 6-24 under 4 DEG C of environment
Hour.The 4% paraformaldehyde fixer, be by 4% paraformaldehyde and 2.5% sucrose, be dissolved in 0.01M PBS prepare and
At.
The brain tissue fixed by after is put into the 0.01M PBS of ice water pre-cooling and rinses 12-24 hours, more renews therebetween
Rinsing liquid 3-5 times thoroughly washes the fixer in tissue.
After rinsing, brain tissue is sequentially placed into 50%, 70% and that 0.5%-0.7%DTT is added in ice water pre-cooling
In 95% ethanol solution, serial dehydration is carried out, it is 1-2 hours each.
After dehydration, by brain tissue be sequentially placed into be added 0.1%-0.2%SBB and 0.7%-1%DTT 50%,
70%, gradient penetration in 85% and 100% GMA solution, each gradient 2-3 hours, wherein 50%, 70% and 85%GMA seeps
The solvent of transparent liquid is 95% ethyl alcohol；Then, then by brain tissue fresh addition 0.1%-0.2%SBB and 0.7%-1% are put into
It is permeated 12-24 hours in 100% resin solution of DTT；It is pre- to be finally putting into addition 0.1%-0.2%SBB and 0.7%-1%DTT
It is permeated 2-3 days in poly- GMA embedding liquid.All steps carry out at 4 DEG C.
Wherein, the above-mentioned 100%GMA used by 52.3%GMA monomer, 3% water, 44.1% butyl methacrylate and
0.6% azobisisoheptonitrile is formulated；The preparation method of pre-polymerization GMA solution is：100%GMA solution is taken to be heated with stirring to 82
DEG C, it when generating a large amount of bubbles, is put into ice-water bath shakes rapidly, be quickly cooled down solution, formed and be similar to viscous syrupy shape
Prepolymer.
The brain tissue that infiltration terminates is put into polymerization mold, fill it up in a mold containing 0.1%-0.2%SBB and
The pre-polymerization GMA of 0.7%-1%DTT embeds liquid, is placed in vacuum drying oven, vacuumizes the oxygen removed in mold, adjusts baking oven
To 38 DEG C, it polymerize 12-24 hours.After polymerization is good, obtained brain tissue embedding sample is the fluorescent sample of high s/n ratio.
TdTomato fluorescence imaging is carried out to embedded brain tissue, gained fluorescent image is referring to Fig. 2.
Using same embodiment, uses the brain tissue of mCherry/dsRed/GFP/BFP label instead, same effect can be obtained
The image of fruit, referring to Fig. 3-6.
Embodiment 2
The mouse heart tissue for taking tdTomato to mark, is put into 4% paraformaldehyde fixer, rear solid under 4 DEG C of environment
Determine 6-24 hours.The 4% paraformaldehyde fixer is dissolved in 0.01M PBS and is matched by 4% paraformaldehyde and 2.5% sucrose
It makes.
The heart tissue fixed by after is put into the 0.01M PBS of ice water pre-cooling and rinses 12-24 hours, more renews therebetween
Rinsing liquid 3-5 times, thoroughly wash the fixer in tissue.
After rinsing, heart tissue is sequentially placed into 50%, 70% and that 0.5%-0.7%DTT is added in ice water pre-cooling
In 95% ethanol solution, serial dehydration is carried out, it is 0.5-1 hours each.
After dehydration, by heart tissue be sequentially placed into containing 0.1%-0.2%SBB and 0.7%-1%DTT 50%,
70%, gradient penetration in 85% and 100% GMA solution, each gradient 1-2 hours, wherein 50%, 70% and 85%GMA seeps
The solvent of transparent liquid is 95% ethyl alcohol；Then, then by heart tissue fresh addition 0.1%-0.2%SBB and 0.7%-1% are put into
It is permeated 6-12 hours in 100% resin solution of DTT；It is finally putting into and the pre- of 0.1%-0.2%SBB and 0.7%-1%DTT is added
It is permeated 1-2 days in poly- GMA embedding liquid.All steps carry out at 4 DEG C.
Wherein, the above-mentioned 100%GMA used by 52.3%GMA monomer, 3% water, 44.1% butyl methacrylate and
0.6% azobisisoheptonitrile is formulated；The preparation method of pre-polymerization GMA solution is：100%GMA solution is taken to be heated with stirring to 82
DEG C, it when generating a large amount of bubbles, is put into ice-water bath shakes rapidly, be quickly cooled down solution, formed and be similar to viscous syrupy shape
Prepolymer.
The heart tissue that infiltration terminates is put into polymerization mold, fill it up in a mold containing 0.1%-0.2%SBB and
The pre-polymerization GMA of 0.7%-1%DTT embeds liquid, is placed in vacuum drying oven, vacuumizes the oxygen removed in mold, adjusts baking oven
To 38 DEG C, it polymerize 12-24 hours.After polymerization is good, obtained heart tissue embedding sample is the fluorescent sample of high s/n ratio.
TdTomato fluorescence imaging is carried out to embedded heart tissue, gained fluorescent image is referring to Fig. 7.
Embodiment 3
Take red fluorescence element mark rat spinal tissues, be put into 4% paraformaldehyde fixer, under 4 DEG C of environment after
It is 6-24 hours fixed.The 4% paraformaldehyde fixer is dissolved in 0.01M PBS by 4% paraformaldehyde and 2.5% sucrose
It is formulated.
The myeloid tissue fixed by after is put into the 0.01M PBS of ice water pre-cooling and rinses 6-12 hours, more renews therebetween
Rinsing liquid 3-5 times, thoroughly wash the fixer in tissue.
After rinsing, myeloid tissue is sequentially placed into 50%, 70% and that 0.5%-0.7%DTT is added in ice water pre-cooling
In 95% ethanol solution, serial dehydration is carried out, it is 0.5-1 hours each.
After dehydration, by myeloid tissue be sequentially placed into containing 0.1%-0.2%SBB and 0.7%-1%DTT 50%,
70%, gradient penetration in 85% and 100% GMA solution, each gradient 1-2 hours, wherein 50%, 70% and 85%GMA seeps
The solvent of transparent liquid is 95% ethyl alcohol；Then, then by myeloid tissue fresh addition 0.1%-0.2%SBB and 0.7%-1% are put into
It is permeated 6-12 hours in 100% resin solution of DTT；It is finally putting into and 0.1%-0.2%SBB and 0.7%-1%DTT pre-polymerization is added
It is permeated 1-2 days in GMA embedding liquid.All steps carry out at 4 DEG C.
Wherein, the above-mentioned 100%GMA used by 52.3%GMA monomer, 3% water, 44.1% butyl methacrylate and
0.6% azobisisoheptonitrile is formulated；The preparation method of pre-polymerization GMA solution is：100%GMA solution is taken to be heated with stirring to 82
DEG C, it when generating a large amount of bubbles, is put into ice-water bath shakes rapidly, be quickly cooled down solution, formed and be similar to viscous syrupy shape
Prepolymer.
The myeloid tissue that infiltration terminates is put into polymerization mold, fill it up in a mold containing 0.1%-0.2%SBB and
The pre-polymerization GMA of 0.7%-1%DTT embeds liquid, is placed in vacuum drying oven, vacuumizes the oxygen removed in mold, adjusts baking oven
To 38 DEG C, it polymerize 12-24 hours.After polymerization is good, obtained myeloid tissue's embedding sample is the fluorescent sample of high s/n ratio.
Embedded myeloid tissue is imaged, gained fluorescent image is referring to Fig. 8.
Using same embodiment, the biological tissue of other fluorescent markers is used instead, the image of effect same can be obtained, join
According to Fig. 3-8.
The above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
Limitation of the invention, protection scope of the present invention should be defined by the scope defined by the claims..For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, several improvements and modifications can also be made, these change
It also should be regarded as protection scope of the present invention into retouching.
Claims (10)
1. a kind of multicolor fluorescence marker samples plasticity embedding method, which is characterized in that including：
Biological sample is pre-processed；
Pretreated biological sample is sequentially placed into the alcoholic solution of the gradient concentration of 0.5-0.7wt% DTT and is permeated,
Each concentration 0.5-3 hours, then continue infiltration 6-12 hours, the various concentration in the resin solution of different quality concentration
Resin solution include 0.1-0.2wt% SBB and 0.7-1wt% DTT；
The fluorescent marker that is embedded after being embedded, being heated to the biological sample after gradient penetration with resin penetration liquid
Sample.
2. the method according to claim 1, wherein it is described to biological sample carry out pretreatment include：
Biological sample is fixed 12-24 hours in paraformaldehyde fixer；
Biological sample after fixation is rinsed 6-12 hours in rinsing liquid；
By the biological sample after the rinsing be sequentially placed into ice water precooling 50%, 70% and 95% or 100% ethanol solution in into
Row serial dehydration, 5 minutes to 2 hours every time.
3. the method according to claim 1, wherein
The mass concentration of the resin solution of the various concentration is 40% ~ 60%, 60% ~ 70%, 70 ~ 90% and 90% ~ 100%.
4. according to the method described in claim 3, it is characterized in that, the mass concentration of the resin solution of the various concentration is distinguished
It is 50%, 70%, 85% and 100%.
5. the method according to claim 1, wherein the mass concentration of the gradient concentration alcoholic solution is respectively
50%, 70%, 85% and 100%.
6. the method according to claim 1, wherein it is described with resin penetration liquid to described after gradient penetration
Biological sample is embedded specially：
Biological sample after gradient penetration is placed in imbedded mold, is then full of resin penetration liquid in a mold.
7. according to the method described in claim 5, it is characterized in that, the resin in the resin penetration liquid and resin solution is specific
For GMA resin.
8. the method according to claim 1, wherein the fluorescent marker sample embedded after heating is specially：
The filled mold is heated 12 to 24 hours at 35 DEG C -40 DEG C.
9. the method according to claim 1, wherein the solvent of the resin solution is 95% ethyl alcohol.
10. method described according to claim 1 ~ 9 any one claims, which is characterized in that the biological sample is selected from
The animal tissue of fluorescence can be emitted.
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| CN109781493A (en) * | 2018-12-26 | 2019-05-21 | 华中科技大学苏州脑空间信息研究院 | It is a kind of can long-time continuous fine upper cut imaging biological tissue's hydrogel embedding method |
| CN109991055A (en) * | 2019-03-28 | 2019-07-09 | 华中科技大学苏州脑空间信息研究院 | The plasticity embedding method of multicolor fluorescence dyestuff and fluorescent protein labeling sample |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109781493A (en) * | 2018-12-26 | 2019-05-21 | 华中科技大学苏州脑空间信息研究院 | It is a kind of can long-time continuous fine upper cut imaging biological tissue's hydrogel embedding method |
| CN109991055A (en) * | 2019-03-28 | 2019-07-09 | 华中科技大学苏州脑空间信息研究院 | The plasticity embedding method of multicolor fluorescence dyestuff and fluorescent protein labeling sample |
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