CN110361242A - It is a kind of for the fixer of eyeball tissue and the preprocess method of eyeball tissue film-making - Google Patents
It is a kind of for the fixer of eyeball tissue and the preprocess method of eyeball tissue film-making Download PDFInfo
- Publication number
- CN110361242A CN110361242A CN201910749874.XA CN201910749874A CN110361242A CN 110361242 A CN110361242 A CN 110361242A CN 201910749874 A CN201910749874 A CN 201910749874A CN 110361242 A CN110361242 A CN 110361242A
- Authority
- CN
- China
- Prior art keywords
- eyeball
- fixer
- eyeball tissue
- tissue
- alcohol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 210000005252 bulbus oculi Anatomy 0.000 title claims abstract description 125
- 238000000034 method Methods 0.000 title claims abstract description 61
- 210000001519 tissue Anatomy 0.000 claims abstract description 83
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 54
- 210000001742 aqueous humor Anatomy 0.000 claims abstract description 23
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 21
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims abstract description 20
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 18
- 229960000935 dehydrated alcohol Drugs 0.000 claims abstract description 18
- 239000000644 isotonic solution Substances 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000004202 carbamide Substances 0.000 claims abstract description 10
- 239000001103 potassium chloride Substances 0.000 claims abstract description 10
- 235000011164 potassium chloride Nutrition 0.000 claims abstract description 10
- 239000011780 sodium chloride Substances 0.000 claims abstract description 9
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 38
- 239000012188 paraffin wax Substances 0.000 claims description 36
- 230000008014 freezing Effects 0.000 claims description 14
- 238000007710 freezing Methods 0.000 claims description 14
- 230000018044 dehydration Effects 0.000 claims description 7
- 238000006297 dehydration reaction Methods 0.000 claims description 7
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 claims description 2
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims 3
- -1 30 min of 1 h Chemical compound 0.000 claims 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- 238000005660 chlorination reaction Methods 0.000 claims 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 210000001525 retina Anatomy 0.000 abstract description 10
- 238000013467 fragmentation Methods 0.000 abstract description 5
- 238000006062 fragmentation reaction Methods 0.000 abstract description 5
- 230000007170 pathology Effects 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 4
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 2
- 230000001575 pathological effect Effects 0.000 abstract description 2
- 238000012545 processing Methods 0.000 description 12
- 238000007796 conventional method Methods 0.000 description 8
- 238000004043 dyeing Methods 0.000 description 6
- 229930040373 Paraformaldehyde Natural products 0.000 description 5
- 229920002866 paraformaldehyde Polymers 0.000 description 5
- 238000005520 cutting process Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 108010077544 Chromatin Proteins 0.000 description 3
- 206010038848 Retinal detachment Diseases 0.000 description 3
- 210000003483 chromatin Anatomy 0.000 description 3
- 238000005336 cracking Methods 0.000 description 3
- 229960004756 ethanol Drugs 0.000 description 3
- 210000000633 nuclear envelope Anatomy 0.000 description 3
- 230000004264 retinal detachment Effects 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 238000004040 coloring Methods 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000003161 choroid Anatomy 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 210000003786 sclera Anatomy 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
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
- G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- 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
-
- 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
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
-
- 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
- G01N2001/305—Fixative compositions
Abstract
The invention belongs to pathology reagent technique fields, and in particular to a kind of for the fixer of eyeball tissue and the preprocess method of eyeball tissue film-making.The each component of the fixer presses volume percentage are as follows: aqueous humor isotonic solution 20~50%, acetic acid 1%~10%, formaldehyde 1~15%, dehydrated alcohol 30%~50%, pure water 10~20%；The aqueous humor isotonic solution is made of sodium chloride, urea, potassium chloride and pure water, and the concentration of sodium chloride is 136~154mM, and the concentration of urea is 10~15mM, and the concentration of potassium chloride is 1~3mM.The integrality that can guarantee the morphosis of eyeball using fixer of the present invention and preprocess method film-making can prevent the fragmentation of retina during film-making and fall off, and to the subsequent morphology of eyeball, histology, pathological research has actively and profound significance.
Description
Technical field
The invention belongs to pathology reagent technique fields, and in particular to a kind of fixer and eyeball group for eyeball tissue
Weave the preprocess method of piece.
Background technique
The precise structure of eyeball is complicated, and the soft or hard degree of each layer tissue of wall of eyeball differs greatly, and retina, choroid, Gong
Connectivity is poor between film.In fixation procedure, conventional fixer is difficult to penetrate fine and close sclera, leads to not using existing
Paraffin section method prepares eyeball paraffin section.Then existing frequently-used paraformaldehyde, which is first fixed, uses ethanol dehydration, still
Since the shrinking percentage of each layer can generate tissue separation, especially causes the separation of retina and fall off.
Further, since there is the pressure aqueous humor and vitreum maintained inside eyeball in eyeball, which in turns increases eyeballs
The difficulty of film-making, if operator directly removes aqueous humor and vitreum by slightly cutting, it is easy to lead to notch out-of-flatness, even
Occur to destroy eyeball structure, leads to retinal detachment and fragmentation.
Summary of the invention
The present invention is in view of the deficiencies of the prior art, and it is an object of the present invention to provide a kind of fixer and eyeball for eyeball tissue
Organize the preprocess method of film-making.
For achieving the above object, the technical scheme adopted by the invention is as follows:
A kind of fixer for eyeball tissue, is made of aqueous humor isotonic solution, acetic acid, formaldehyde, dehydrated alcohol and pure water, institute
It states aqueous humor isotonic solution to be made of sodium chloride, urea, potassium chloride and pure water, each component of the fixer presses volume percentage
Are as follows: aqueous humor isotonic solution 20~50%, acetic acid 1%~10%, formalin 1~15%, dehydrated alcohol 30%~50%, pure water 10
~20%.
In above scheme, the concentration of sodium chloride is 136~154mM in the aqueous humor isotonic solution, the concentration of urea is 10~
15mM, the concentration of potassium chloride are 1~3mM.
In above scheme, the volumetric concentration of the formalin is 37~40%.
A kind of preprocess method of eyeball tissue film-making, includes the following steps:
(1) using the fixed eyeball tissue of the fixer, then by fixed eyeball tissue frozen section embedding medium
Embedding；
(2) eyeball tissue after step (1) embedding is freezed, eyeball tissue is made to be hardened；Then to the eyeball tissue of freezing
It is slightly cut, removes the aqueous humor and vitreum in eyeball tissue；
(3) eyeball tissue that step (2) eliminates aqueous humor and vitreum is fixed with the fixer again；Fixation terminates
Eyeball tissue carries out to gradient alcohol dehydration, dimethylbenzene is transparent, paraffin waxdip embeds again afterwards.
In above scheme, step (1) the frozen section embedding medium is OCT embedding medium.
In above scheme, step (1) time using the fixed eyeball tissue of fixer is 22~for 24 hours.
In above scheme, the freezing of step (2) described eyeball tissue and thick cut carry out all on freezing microtome.
In above scheme, the temperature of step (2) the freezing eyeball tissue is -20 DEG C to -25 DEG C.
In above scheme, step (2) form slightly cut are as follows: the sagittal plane direction along eyeball tissue cuts the eyeball
The 1/4 to 1/3 of tissue.
In above scheme, step (3) described eyeball tissue is again 24~48h with the fixer regular time.
In above scheme, the detailed process of step (3) described gradient alcohol dehydration are as follows: 75% alcohol 4h, 85% alcohol 2h,
90% alcohol 2h, 95% alcohol 1h, dehydrated alcohol I 30min, dehydrated alcohol II30min.
In above scheme, the transparent detailed process of the dimethylbenzene are as follows: dimethylbenzene 5~10min of I, dimethylbenzene II 5~
10min。
In above scheme, the detailed process of the paraffin waxdip embedding are as follows: paraffin I 1h, paraffin II 1h, paraffin III 1h
Beneficial effects of the present invention:
(1) fixer of the present invention quickly can either be infiltrated into fully in eyeball tissue, and will not make each of eyeball
Structure sheaf expands, and is conducive to the structural intergrity for keeping eyeball；Preprocess method of the present invention, which passes through, first freezes eyeball,
Then eyeball is carried out with frozen section knife cutting repairing, to remove the aqueous humor and vitreum inside eyeball, eyeball tissue is through cold
Tissue, which is hardened, after freezing cuts machine maintenance convenient for ice below and cuts, and than repairing by hand, face earnestly is smooth to prevent the falling off of retina, broken with this
It splits, and eyeball is prevented to deform in sample making course.
(2) eyeball slicing treatment, each layer tissue structure of eyeball are carried out using fixer of the present invention and preprocess method
Completely, no obvious structure changes, and has no falling off and separating, each layer tissue of eyeball no crack clear in structure for retina；Slice thick
It is thin uniform, there is no flake phenomenon；No cracking deforms between histocyte, and cell-free excess shrinkage and expansion have no the self-dissolving of tissue；
Entoblast, nuclear membrane are clear, and chromatin is uniform, bright color, and color contrast is obvious；It can preferably guarantee eyeball shape, prevent
It the fragmentation of retina and falls off during film-making, to the subsequent morphology of eyeball, histology, pathological research has positive and far-reaching
Meaning.
Detailed description of the invention
Fig. 1 is the HE colored graph of the eyeball slice obtained using conventional method.
Fig. 2 is that embodiment 1 is contaminated using the HE of the eyeball slice prepared after fixer of the invention and preprocess method processing
Chromatic graph.
Fig. 3 is that embodiment 2 is contaminated using the HE of the eyeball slice prepared after fixer of the invention and preprocess method processing
Chromatic graph.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is furture elucidated, but it is of the invention
Content is not limited solely to the following examples.
Embodiment 1
The ingredient of the fixer of eyeball tissue is as follows: aqueous humor isotonic solution 35mL, formaldehyde 10mL, acetic acid 5mL, dehydrated alcohol
37.5mL, pure water 12.5mL.The aqueous humor isotonic solution is made of sodium chloride, urea, potassium chloride and pure water, and the concentration of sodium chloride is
137.9mM, the concentration of urea are 13.43mM, and the concentration of potassium chloride is 2.67mM.Aforesaid liquid is mixed, it is heavy to obtain colourless nothing
The solution in shallow lake is the fixer of the present embodiment.
A kind of preprocess method of eyeball tissue film-making, includes the following steps:
(1) using the fixed eyeball tissue 22h of the fixer；
(2) eyeball tissue fixed using OCT embedding medium embedding step (1)；
(3) eyeball tissue that step (2) embed is placed at -20 DEG C and is freezed；
(4) eyeball tissue for the freezing for obtaining step (3) is under the conditions of on freezing microtome -20 DEG C, along sagittal plane side
To cutting the 1/4 of the eyeball tissue, the aqueous humor and vitreum in the eyeball tissue are removed；
(5) by step (4) treated eyeball tissue return in the fixer it is fixed for 24 hours；
(6) eyeball tissue for fixing step (5) uses gradient alcohol dehydration, and detailed process is 75% alcohol 4h,
85% alcohol 2h, 90% alcohol 2h, 95% alcohol 1h, dehydrated alcohol I 30min, dehydrated alcohol II 30min；
(7) it is transparent to reuse dimethylbenzene, detailed process are as follows: 5~10min of dimethylbenzene 5~10min of I, dimethylbenzene II；
(8) embedding of paraffin waxdip, detailed process are as follows: paraffin I 1h, paraffin II 1h, paraffin III 1h are reused.
Finally by the eyeball tissue embedded section by waxdip processing, paraffin section is obtained.Pre- through this embodiment it will locate
The paraffin section that reason method obtains carries out HE dyeing, and sealing, processing of taking pictures is to get effect shown in Fig. 2.(note: HE staining procedure is
Colouring method well-known to those skilled in the art).
HE dyeing is carried out with the eyeball paraffin section that conventional tabletting method obtains as control.It is described to be made of conventional method
The process of eyeball slice are as follows:
(1) for 24 hours using the fixed eyeball tissue of 4% paraformaldehyde；
(2) eyeball tissue of step (1) after fixed is dehydrated with graded ethanol, detailed process are as follows: 75% alcohol 4h,
85% alcohol 2h, 90% alcohol 2h, 95% alcohol 1h, dehydrated alcohol I 30min, dehydrated alcohol II 30min；
(3) transparent using dimethylbenzene, detailed process are as follows: dimethylbenzene I 5-10min, dimethylbenzene II 5-10min；
(4) it is embedded using paraffin waxdip, detailed process are as follows: paraffin I 1h, paraffin II 1h, paraffin III 1h；
By the eyeball tissue embedded section by waxdip processing, production eyeball slice is then subjected to HE dyeing, sealing is clapped
According to processing to get effect shown in Fig. 1.
Fig. 1 is the HE colored graph of the eyeball slice obtained using conventional method, can be seen that use from result shown in Fig. 1
Paraformaldehyde is fixed and is used in the eyeball paraffin section of conventional method film-making, and retinal detachment, eyeball structure is badly deformed.
Fig. 2 is the HE colored graph using the eyeball slice prepared after fixer of the invention and preprocess method processing.From
As can be seen that each layer tissue structural integrity of eyeball in Fig. 2, no obvious structure changes, and has no falling off and separating, eyeball for retina
Each layer tissue no crack clear in structure；It is uniform to be sliced thickness, does not have flake phenomenon；No cracking deforms between histocyte, cell-free
Excess shrinkage and expansion have no the self-dissolving of tissue；Entoblast, nuclear membrane are clear, and chromatin is uniform, bright color, color contrast
Obviously.
The comparison of Fig. 2 and Fig. 1, it can be deduced that fixation and film-making for eyeball, the present invention in eyeball fixer and match
The preprocess method of the eyeball tissue film-making of set can be preferably than conventional tabletting method known to those skilled in the art of the present technique
The form for guaranteeing eyeball prevents the fragmentation of retina during film-making and falls off, to the subsequent morphology of eyeball, histology, pathology
Research has actively and profound significance.
Embodiment 2
The ingredient of the fixer of eyeball tissue is as follows: aqueous humor isotonic solution 40mL, formaldehyde 7mL, acetic acid 3mL, dehydrated alcohol
30mL, pure water 20mL.The aqueous humor isotonic solution is made of sodium chloride, urea, potassium chloride and pure water, and wherein sodium chloride concentration is
150mM, urea concentration 3mM, potassium chloride concentration 1mM.Aforesaid liquid is mixed, the solution for obtaining colourless no precipitating is this
The fixer of embodiment.
A kind of preprocess method of eyeball tissue film-making, includes the following steps:
(1) for 24 hours using the fixed eyeball tissue of the fixer；
(2) eyeball tissue fixed using OCT embedding medium embedding step (1)；
(3) eyeball tissue that step (2) embed is placed in -25 DEG C of freezings；
(4) eyeball tissue for the freezing for obtaining step (3) is under the conditions of on freezing microtome -25 DEG C, along sagittal plane side
To cutting the 1/3 of the eyeball tissue, the aqueous humor and vitreum in the eyeball tissue are removed；
(5) step (4) treated eyeball tissue is returned in the fixer and fixes 48h；
(6) eyeball tissue for fixing step (5) uses gradient alcohol dehydration, detailed process are as follows: 75% alcohol 4h,
85% alcohol 2h, 90% alcohol 2h, 95% alcohol 1h, dehydrated alcohol I 30min, dehydrated alcohol II 30min；
(7) it is transparent to reuse dimethylbenzene, detailed process are as follows: 5~10min of dimethylbenzene 5~10min of I, dimethylbenzene II；
(8) embedding of paraffin waxdip, detailed process are as follows: paraffin I 1h, paraffin II 1h, paraffin III 1h are reused；
Finally by the eyeball tissue embedded section by waxdip processing, paraffin section is obtained.Pre- through this embodiment it will locate
The paraffin section that reason method obtains carries out HE dyeing, and sealing, processing of taking pictures is to get effect shown in Fig. 3.(note: HE staining procedure is
Colouring method well-known to those skilled in the art).
HE dyeing is carried out with the eyeball paraffin section that conventional tabletting method obtains as control.It is described to be made of conventional method
The process of eyeball slice are as follows:
(1) for 24 hours using the fixed eyeball tissue of 4% paraformaldehyde；
(2) eyeball tissue of step (1) after fixed is dehydrated with graded ethanol, detailed process are as follows: 75% alcohol 4h,
85% alcohol 2h, 90% alcohol 2h, 95% alcohol 1h, dehydrated alcohol I 30min, dehydrated alcohol II 30min；
(3) transparent using dimethylbenzene, detailed process are as follows: 5~10min of dimethylbenzene 5~10min of I, dimethylbenzene II；
(4) it is embedded using paraffin waxdip, detailed process are as follows: paraffin I 1h, paraffin II 1h, paraffin III 1h；
By the eyeball tissue embedded section by waxdip processing.Conventional method production eyeball slice is subjected to HE dyeing, envelope
Gu processing of taking pictures is to get effect shown in Fig. 1.
Fig. 1 is the HE colored graph of the eyeball slice obtained using conventional method, can be seen that use from result shown in Fig. 1
Paraformaldehyde is fixed and is used in the eyeball paraffin section of conventional method film-making, and retinal detachment, eyeball structure is badly deformed.
Fig. 3 is the HE colored graph using the eyeball slice prepared after fixer of the invention and preprocess method processing.From
As can be seen that each layer tissue structural integrity of eyeball in Fig. 3, no obvious structure changes, and has no falling off and separating, eyeball for retina
Each layer tissue no crack clear in structure；It is uniform to be sliced thickness, does not have flake phenomenon；No cracking deforms between histocyte, cell-free
Excess shrinkage and expansion have no the self-dissolving of tissue；Entoblast, nuclear membrane are clear, and chromatin is uniform, bright color, color contrast
Obviously.
The comparison of Fig. 3 and Fig. 1, it can be deduced that fixation and film-making for eyeball, the present invention in eyeball fixer and match
The preprocess method of the eyeball tissue film-making of set can be preferably than conventional tabletting method known to those skilled in the art of the present technique
The form for guaranteeing eyeball prevents the fragmentation of retina during film-making and falls off, to the subsequent morphology of eyeball, histology, pathology
Research has actively and profound significance.
Obviously, above-described embodiment is only intended to clearly illustrate made example, and is not the limitation to embodiment.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of variation or
It changes.There is no necessity and possibility to exhaust all the enbodiments.And the obvious variation or change therefore amplified
It moves within still in the protection scope of the invention.
Claims (10)
1. a kind of fixer for eyeball tissue, which is characterized in that the fixer is by aqueous humor isotonic solution, acetic acid, formaldehyde, nothing
Water-ethanol and pure water composition, the aqueous humor isotonic solution is made of sodium chloride, urea, potassium chloride and pure water, the fixer it is each
Component presses volume percentage are as follows: aqueous humor isotonic solution 20 ~ 50%, acetic acid 1% ~ 10%, formaldehyde 1 ~ 15%, dehydrated alcohol 30% ~ 50%, pure
Water 10 ~ 20%.
2. being used for the fixer of eyeball tissue according to claim 1, which is characterized in that chlorination in the aqueous humor isotonic solution
The concentration of sodium is 136 ~ 154 mM, and the concentration of urea is 10 ~ 15 mM, and the concentration of potassium chloride is 1 ~ 3 mM.
3. being used for the fixer of eyeball tissue according to claim 1, which is characterized in that the volume of the formalin is dense
Degree is 37 ~ 40%.
4. carrying out the preprocess method of eyeball tissue film-making using any fixer of claim 1 ~ 3, which is characterized in that packet
Include following steps:
(1) eyeball tissue is fixed using the fixer, then fixed eyeball tissue frozen section embedding medium is embedded；
(2) eyeball tissue after step (1) embedding is freezed, eyeball tissue is made to be hardened；Then the eyeball tissue of freezing is carried out
It slightly cuts, removes the aqueous humor and vitreum in eyeball tissue；
(3) eyeball tissue that step (2) eliminates aqueous humor and vitreum is fixed with the fixer again；After fixation again
Eyeball tissue carries out to gradient alcohol dehydration, dimethylbenzene is transparent, the embedding of paraffin waxdip.
5. preprocess method according to claim 4, which is characterized in that step (1) the frozen section embedding medium is OCT
Embedding medium, the time using the fixed eyeball tissue of fixer is 22 ~ 24 h.
6. preprocess method according to claim 4, which is characterized in that the freezing of step (2) described eyeball tissue and thick
It cuts and is carried out all on freezing microtome；The form slightly cut are as follows: cut the eyeball group along the sagittal plane direction of eyeball tissue
1/4 to 1/3 knitted.
7. preprocess method according to claim 4, which is characterized in that the temperature of step (2) the freezing eyeball tissue
It is -20 DEG C to -25 DEG C；Step (3) described eyeball tissue is again 24 ~ 48 h with the fixer regular time.
8. preprocess method according to claim 4, which is characterized in that step (3) described gradient alcohol dehydration it is specific
Process are as follows: 75% alcohol, 4 h, 85% alcohol, 2 h, 90% alcohol, 2 h, 95% alcohol, 30 min of 1 h, dehydrated alcohol I, dehydrated alcohol
II 30 min。
9. preprocess method according to claim 4, which is characterized in that the transparent detailed process of the dimethylbenzene are as follows: two
5 ~ 10 min of toluene I 5 ~ 10 min, dimethylbenzene II.
10. preprocess method according to claim 4, which is characterized in that the detailed process of the paraffin waxdip embedding are as follows:
1 h of paraffin I 1 h, paraffin II 1 h, paraffin III.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910749874.XA CN110361242B (en) | 2019-08-14 | 2019-08-14 | Fixing liquid for eyeball tissue and pretreatment method for preparing eyeball tissue slices |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910749874.XA CN110361242B (en) | 2019-08-14 | 2019-08-14 | Fixing liquid for eyeball tissue and pretreatment method for preparing eyeball tissue slices |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110361242A true CN110361242A (en) | 2019-10-22 |
| CN110361242B CN110361242B (en) | 2022-01-18 |
Family
ID=68223757
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910749874.XA Active CN110361242B (en) | 2019-08-14 | 2019-08-14 | Fixing liquid for eyeball tissue and pretreatment method for preparing eyeball tissue slices |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110361242B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111238901A (en) * | 2020-03-04 | 2020-06-05 | 四川省人民医院 | Manufacturing method of improved mouse eyeball frozen section |
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
| CN112781960A (en) * | 2021-02-05 | 2021-05-11 | 四川省人民医院 | Method for making eye frozen section |
Citations (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5460797A (en) * | 1991-05-08 | 1995-10-24 | Streck Laboratories, Inc. | Method for fixing tissues and cells for analysis using oxazolidine compounds |
| WO2001063294A2 (en) * | 2000-02-24 | 2001-08-30 | Oxford Glycosciences (Uk) Limited | Diagnosis of bipolar affective disorder (bad) and unipolar depression |
| US20020009453A1 (en) * | 2000-06-16 | 2002-01-24 | Symphogen A/S | Polyclonal antibody composition for treating allergy |
| US20030032773A1 (en) * | 2000-02-24 | 2003-02-13 | Herath Herath Mudiyanselage Athula Chandrasiri | Proteins, genes and their use for diagnosis and treatment of bipolar affective disorder (BAD) and unipolar depression |
| WO2003066560A1 (en) * | 2002-02-05 | 2003-08-14 | Biesecker James L | Stabilized powdered formaldehyde |
| WO2003096876A2 (en) * | 2001-04-01 | 2003-11-27 | Sommer Phillipe L | Continuously operational diagnostic device |
| US20040110938A1 (en) * | 2000-02-24 | 2004-06-10 | Parekh Rajesh Bhikhu | Proteins, genes and their use for diagnosis and treatment of schizophrenia |
| CN1738641A (en) * | 2002-12-06 | 2006-02-22 | 思罗姆-X股份有限公司 | Pharmacological vitreolysis |
| US20090035761A1 (en) * | 1999-12-20 | 2009-02-05 | University Of Southern California | Method for isolation of DNA, RNA and proteins from formalin-fixed paraffin-embedded tissue specimens |
| EP2052080A1 (en) * | 2006-08-17 | 2009-04-29 | Department Of Biotechnology | Resistance of plants to biotic and abiotic stresses by overexpression of protochlorophyllide oxidoreductase c and its isoforms |
| US20110105726A1 (en) * | 1997-03-07 | 2011-05-05 | Human Genome Sciences, Inc. | Antibodies to hsdek49 polypeptides |
| WO2013028807A2 (en) * | 2011-08-22 | 2013-02-28 | Somalogic, Inc. | Renal cell carcinoma biomarkers and uses thereof |
| CN103083298A (en) * | 2012-11-20 | 2013-05-08 | 蒋春红 | Application of salvianolic acid A composition in preparing medicines for protecting ischemic brain tissue damage |
| CN103083297A (en) * | 2012-11-20 | 2013-05-08 | 蒋春红 | Application of salvianolic acid A composition in preparing medicines for improving neural function symptom after cerebral ischemia |
| CN103471903A (en) * | 2013-09-29 | 2013-12-25 | 吴天骥 | Method for preparing stationary liquid for slicing and application of stationary liquid for slicing |
| WO2014152211A1 (en) * | 2013-03-14 | 2014-09-25 | Moderna Therapeutics, Inc. | Formulation and delivery of modified nucleoside, nucleotide, and nucleic acid compositions |
| CN104663647A (en) * | 2013-11-27 | 2015-06-03 | 深圳先进技术研究院 | Heart tissue fixing agent as well as preparation method and application of heart tissue fixing agent |
| CN105241686A (en) * | 2015-08-10 | 2016-01-13 | 河南科技大学 | Preparation method of retina microscopic tissue slice of hynobiidae animals |
| CN105571916A (en) * | 2015-12-24 | 2016-05-11 | 四川省农业科学院水产研究所 | Acipenser dabryanus oosperm tissue slicing method |
| CN105560270A (en) * | 2016-03-05 | 2016-05-11 | 广东宏盈科技有限公司 | Lubricating liquid for corneal contact lens |
| CN106908294A (en) * | 2017-03-24 | 2017-06-30 | 漯河医学高等专科学校 | A kind of processing method of Pathologic specimen |
| CN110426259A (en) * | 2019-08-14 | 2019-11-08 | 武汉赛维尔生物科技有限公司 | Polyethylene glycol is used for the application of animal tissue sections grease dyeing |
| CN110823663A (en) * | 2019-10-09 | 2020-02-21 | 中国人民解放军总医院第五医学中心 | Method for manufacturing pathological section of animal eyeball |
| CN110887719A (en) * | 2019-12-20 | 2020-03-17 | 云南中医药大学 | Method for processing tissue sample of histopathology |
| CN111238901A (en) * | 2020-03-04 | 2020-06-05 | 四川省人民医院 | Manufacturing method of improved mouse eyeball frozen section |
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
-
2019
- 2019-08-14 CN CN201910749874.XA patent/CN110361242B/en active Active
Patent Citations (27)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5460797A (en) * | 1991-05-08 | 1995-10-24 | Streck Laboratories, Inc. | Method for fixing tissues and cells for analysis using oxazolidine compounds |
| US20110105726A1 (en) * | 1997-03-07 | 2011-05-05 | Human Genome Sciences, Inc. | Antibodies to hsdek49 polypeptides |
| US20090035761A1 (en) * | 1999-12-20 | 2009-02-05 | University Of Southern California | Method for isolation of DNA, RNA and proteins from formalin-fixed paraffin-embedded tissue specimens |
| US20040110938A1 (en) * | 2000-02-24 | 2004-06-10 | Parekh Rajesh Bhikhu | Proteins, genes and their use for diagnosis and treatment of schizophrenia |
| US20030032773A1 (en) * | 2000-02-24 | 2003-02-13 | Herath Herath Mudiyanselage Athula Chandrasiri | Proteins, genes and their use for diagnosis and treatment of bipolar affective disorder (BAD) and unipolar depression |
| WO2001063294A2 (en) * | 2000-02-24 | 2001-08-30 | Oxford Glycosciences (Uk) Limited | Diagnosis of bipolar affective disorder (bad) and unipolar depression |
| US20020009453A1 (en) * | 2000-06-16 | 2002-01-24 | Symphogen A/S | Polyclonal antibody composition for treating allergy |
| WO2003096876A2 (en) * | 2001-04-01 | 2003-11-27 | Sommer Phillipe L | Continuously operational diagnostic device |
| WO2003066560A1 (en) * | 2002-02-05 | 2003-08-14 | Biesecker James L | Stabilized powdered formaldehyde |
| US20150064161A1 (en) * | 2002-12-06 | 2015-03-05 | Thrombogenics Nv | Pharmacological vitreolysis |
| CN1738641A (en) * | 2002-12-06 | 2006-02-22 | 思罗姆-X股份有限公司 | Pharmacological vitreolysis |
| EP2052080A1 (en) * | 2006-08-17 | 2009-04-29 | Department Of Biotechnology | Resistance of plants to biotic and abiotic stresses by overexpression of protochlorophyllide oxidoreductase c and its isoforms |
| WO2013028807A2 (en) * | 2011-08-22 | 2013-02-28 | Somalogic, Inc. | Renal cell carcinoma biomarkers and uses thereof |
| CN103083298A (en) * | 2012-11-20 | 2013-05-08 | 蒋春红 | Application of salvianolic acid A composition in preparing medicines for protecting ischemic brain tissue damage |
| CN103083297A (en) * | 2012-11-20 | 2013-05-08 | 蒋春红 | Application of salvianolic acid A composition in preparing medicines for improving neural function symptom after cerebral ischemia |
| WO2014152211A1 (en) * | 2013-03-14 | 2014-09-25 | Moderna Therapeutics, Inc. | Formulation and delivery of modified nucleoside, nucleotide, and nucleic acid compositions |
| CN103471903A (en) * | 2013-09-29 | 2013-12-25 | 吴天骥 | Method for preparing stationary liquid for slicing and application of stationary liquid for slicing |
| CN104663647A (en) * | 2013-11-27 | 2015-06-03 | 深圳先进技术研究院 | Heart tissue fixing agent as well as preparation method and application of heart tissue fixing agent |
| CN105241686A (en) * | 2015-08-10 | 2016-01-13 | 河南科技大学 | Preparation method of retina microscopic tissue slice of hynobiidae animals |
| CN105571916A (en) * | 2015-12-24 | 2016-05-11 | 四川省农业科学院水产研究所 | Acipenser dabryanus oosperm tissue slicing method |
| CN105560270A (en) * | 2016-03-05 | 2016-05-11 | 广东宏盈科技有限公司 | Lubricating liquid for corneal contact lens |
| CN106908294A (en) * | 2017-03-24 | 2017-06-30 | 漯河医学高等专科学校 | A kind of processing method of Pathologic specimen |
| CN110426259A (en) * | 2019-08-14 | 2019-11-08 | 武汉赛维尔生物科技有限公司 | Polyethylene glycol is used for the application of animal tissue sections grease dyeing |
| CN110823663A (en) * | 2019-10-09 | 2020-02-21 | 中国人民解放军总医院第五医学中心 | Method for manufacturing pathological section of animal eyeball |
| CN110887719A (en) * | 2019-12-20 | 2020-03-17 | 云南中医药大学 | Method for processing tissue sample of histopathology |
| CN111238901A (en) * | 2020-03-04 | 2020-06-05 | 四川省人民医院 | Manufacturing method of improved mouse eyeball frozen section |
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
Non-Patent Citations (15)
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111238901A (en) * | 2020-03-04 | 2020-06-05 | 四川省人民医院 | Manufacturing method of improved mouse eyeball frozen section |
| CN111238901B (en) * | 2020-03-04 | 2023-03-03 | 四川省人民医院 | Manufacturing method of improved mouse eyeball frozen section |
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
| CN112781960A (en) * | 2021-02-05 | 2021-05-11 | 四川省人民医院 | Method for making eye frozen section |
| CN112781960B (en) * | 2021-02-05 | 2024-02-23 | 四川省人民医院 | Method for making frozen eye slices |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110361242B (en) | 2022-01-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110361242A (en) | It is a kind of for the fixer of eyeball tissue and the preprocess method of eyeball tissue film-making | |
| Untersee et al. | Visualization of extracellular lining layer of lung alveoli by freeze-etching | |
| EP3090704B1 (en) | Method for preparing artificial heart valve leaflets | |
| CN106279728B (en) | Preparation method of multiphase mixed linked cross-linked hyaluronic acid | |
| CN111238901B (en) | Manufacturing method of improved mouse eyeball frozen section | |
| CN110823663A (en) | Method for manufacturing pathological section of animal eyeball | |
| Schoenwolf et al. | Descriptive studies of occlusion and reopening of the spinal canal of the early chick embryo | |
| CN109400698B (en) | Preparation method of low endotoxin collagen | |
| CN110132673B (en) | Method for preparing needle mushroom fruiting body tissue slices | |
| CN105037752A (en) | Modified polyvinyl alcohol cornea repair material with collagen in graded distribution and preparation method | |
| Wilson et al. | Differential swelling in compartments of the corneal stroma. | |
| Koay et al. | Fascin expression in cervical normal squamous epithelium, cervical intraepithelial neoplasia, and superficially invasive (stage IA1) squamous carcinoma of the cervix | |
| Banfield et al. | Electronmicrographs of thin sections of Molluscum contagiosum. | |
| CN107057108B (en) | Chitosan material with controllable structure and performance and preparation method thereof | |
| CN112293407A (en) | Method for programmed cryopreservation of ovarian tissues | |
| CN114088493B (en) | Animal eyeball pathological section manufacturing method | |
| CN109646705A (en) | Composite sponge and preparation method thereof and negative pressure drainage dressing, device and Medical Devices | |
| CN204855183U (en) | Eyeball pathologic sampling mould | |
| RU2734280C1 (en) | Method for isolated extraction and fabrication of histological structures of vitreous body | |
| Hyde et al. | Ultrastructure of zoosporogenesis in Phytophthora cinnamomi | |
| CN113702148A (en) | Freezing embedding agent and application thereof in frozen section | |
| CN107976353B (en) | Preparation method of crop tender root paraffin slice | |
| Robert Charles et al. | Lichen striatus: A clinical, histologic and electron microscopic study of an unusual case | |
| RU2538383C1 (en) | Method for performing complete corneal stromectomy | |
| CN112779286A (en) | Construction method and application of Alport syndrome mouse model |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |