CN105910861B - A kind of automatic method of batch dyeing production histotomy - Google Patents
A kind of automatic method of batch dyeing production histotomy Download PDFInfo
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- CN105910861B CN105910861B CN201610228177.6A CN201610228177A CN105910861B CN 105910861 B CN105910861 B CN 105910861B CN 201610228177 A CN201610228177 A CN 201610228177A CN 105910861 B CN105910861 B CN 105910861B
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- sample
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- working fluid
- layer part
- histotomy
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- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000004043 dyeing Methods 0.000 title claims abstract description 17
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 12
- 239000000975 dye Substances 0.000 claims abstract description 19
- 210000004556 brain Anatomy 0.000 claims abstract description 18
- 239000012530 fluid Substances 0.000 claims abstract description 12
- 239000012466 permeate Substances 0.000 claims abstract description 3
- 230000035515 penetration Effects 0.000 claims description 3
- 230000003204 osmotic effect Effects 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 2
- 210000005013 brain tissue Anatomy 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000000653 nervous system 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
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- 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
- G01N1/31—Apparatus therefor
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses the automatic methods that a kind of batch dyes production histotomy, this method comprises: sample to be processed is immersed in the working fluid containing architecture cytoarchitectonic dyestuff by (1)；(2) dye molecule in working fluid permeates from sample surface to sample depths, by after a certain period of time, the shallow-layer part architecture cytoarchitectonic of sample surface is colored；(3) it is sliced using the cutter shallow-layer part dyed to sample；(4) dye molecule in working fluid infiltrates into shallow-layer part architecture cytoarchitectonic to sample depths from slice profile and is colored, and will be controlled by computer, and sample is lifted height identical with slice thickness；(5) repeat the above steps (3)~(4), until entire sample to be cut into the brain piece of architecture cytoarchitectonic dyeing.The method of the present invention, using the naturally osmotic in slice gap dyestuff, avoids manually operated interference in dyeing course, time saving and energy saving, is particularly suitable for batch dyeing, production histotomy.
Description
Technical field
The present invention relates to the production of biological tissue section, and the automation of production histotomy is dyed in particular to a kind of batch
Method belongs to life science technical field.
Background technique
In order to be situated between see or microscopic scale research Different brain region neuromechanism and functional information, researcher usually require by
Brain tissue is cut into 100 microns or thinner of brain piece, recycle label and imaging means to dye these brain pieces piecewise, at
Picture.
Since nervous system is divided with brain area, there are larger differences for the nerve of Different brain region, therefore, to neuromechanism with
When functional information is analyzed, need first to position brain area where it.Currently, the method for realizing brain area positioning is mainly
It is carried out using the otherness of Different brain region architecture cytoarchitectonic distribution, after carrying out architecture cytoarchitectonic dyeing to brain piece, the brain can be obtained
The architecture cytoarchitectonic information of piece, control reference brain map such as " Allen Reference Atlas " compare similar imaging results, with
Brain area where confirmation.However, such acquisition of information, the manual process usually dyed piecewise again after experience slice, i.e.,
Make Mouse brain outer profile size also up to more than 10 millimeters, needs to be cut into tens or hundreds of brain pieces.Biological experiment generally requires
Many experiments are carried out to obtain statistical rule, eliminate individual difference bring error.Therefore, this inefficient laborious to cut
Piece, colouring method are not able to satisfy the needs of existing research.Urgently develop a kind of side for automating batch dyeing, production histotomy
Method.
Summary of the invention
Present invention aims to overcome that above-mentioned the deficiencies in the prior art and a kind of batch dyeing production histotomy is provided
Automatic method, method includes the following steps:
(1) sample to be processed is immersed in the working fluid containing architecture cytoarchitectonic dyestuff；
(2) dye molecule in working fluid permeates from sample surface to sample depths, by after a certain period of time, sample table
The shallow-layer part architecture cytoarchitectonic in face is colored；
(3) it is sliced using the cutter shallow-layer part dyed to sample；
(4) dye molecule in working fluid infiltrates into shallow-layer part architecture cytoarchitectonic to sample depths from slice profile and is contaminated
Color will be controlled by computer, and sample is lifted height identical with slice thickness；
(5) repeat the above steps (3)~(4), until entire sample to be cut into the brain piece of architecture cytoarchitectonic dyeing.
In the above-mentioned technical solutions, the sample side is coated with the medium for preventing dyestuff penetration.
The method of the present invention has the advantage that
Dyeing is carried out with slicing processes automation, the operation of the method for the present invention is first thin to the shallow-layer part of sample surface
Born of the same parents construct dyeing, and then the shallow-layer part dyed to sample is sliced, and recycle the operation of above-mentioned " dyeing-slice ", because
And the method for the present invention can be realized by way of automatic control.This method is first sliced the working method dyed piecewise again with original
It compares, improves the efficiency handled piecewise brain piece.Dyeing course utilizes the naturally osmotic in slice gap dyestuff, avoids
Manually operated interference, it is time saving and energy saving.
Present invention is especially suited for batch dyeing, production histotomy, for the extensive experiment number for obtaining Neuroscience Research
According to providing new experimental tool.
Detailed description of the invention
Fig. 1 is the schematic diagram of batch dyeing production histotomy by the method for the invention.
Specific embodiment
The following further describes the present invention in detail with reference to the accompanying drawings and specific embodiments.
As shown in Figure 1, sample 1 is first fixed in sink 3 by the method for the present invention, it is filled in sink 3 containing architecture cytoarchitectonic
The working fluid 2 of dyestuff, sample 1 is flooded completely.The side of sample used in the present invention is coated with the medium for preventing dyestuff penetration, such as
Nail polish, so that dyestuff can not be permeated from the side of sample.After standing enough time, working fluid 2 is permeated into sample 1, so that
The architecture cytoarchitectonic of surface shallow-layer sample is colored.Sample 1 is sliced using cutter 5, so that dyed 4 quilt of shallow-layer part
Cut off sample, the next surface to be dyed, slice of exposure.Dye molecule in working fluid is from slice profile to sample depths
It infiltrates into shallow-layer part architecture cytoarchitectonic to be colored, by computer controlled automatic, sample is lifted height identical with slice thickness
Degree.Dyeing-slicing processes circulation can automate and repeat, until sample is cut.
Claims (2)
1. a kind of automatic method of batch dyeing production histotomy characterized by comprising
(1) sample to be processed is immersed in the working fluid containing architecture cytoarchitectonic dyestuff；
(2) dye molecule in working fluid permeates from sample surface to sample depths, by after a certain period of time, sample surface
Shallow-layer part architecture cytoarchitectonic is colored；
(3) it is sliced using the cutter shallow-layer part dyed to sample；
(4) dye molecule in working fluid infiltrates into shallow-layer part architecture cytoarchitectonic to sample depths from slice profile and is colored,
It will be controlled by computer, sample is lifted height identical with slice thickness；
(5) repeat the above steps (3)~(4), until entire sample to be cut into the brain piece of architecture cytoarchitectonic dyeing.
2. the automatic method of batch dyeing production histotomy according to claim 1, it is characterised in that: the sample side
Face is coated with the medium for preventing dyestuff penetration.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610228177.6A CN105910861B (en) | 2016-04-13 | 2016-04-13 | A kind of automatic method of batch dyeing production histotomy |
| PCT/CN2017/079847 WO2017177870A1 (en) | 2016-04-13 | 2017-04-10 | Automated batch method for staining and making tissue slices |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610228177.6A CN105910861B (en) | 2016-04-13 | 2016-04-13 | A kind of automatic method of batch dyeing production histotomy |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105910861A CN105910861A (en) | 2016-08-31 |
| CN105910861B true CN105910861B (en) | 2018-12-11 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610228177.6A Active CN105910861B (en) | 2016-04-13 | 2016-04-13 | A kind of automatic method of batch dyeing production histotomy |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN105910861B (en) |
| WO (1) | WO2017177870A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105910861B (en) * | 2016-04-13 | 2018-12-11 | 武汉沃亿生物有限公司 | A kind of automatic method of batch dyeing production histotomy |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101458182A (en) * | 2008-12-22 | 2009-06-17 | 华中科技大学 | Method for producing brain sample of small animal |
| CN201532333U (en) * | 2009-11-09 | 2010-07-21 | 徐华 | Histiocyte section totally-enclosed full-automatic dyeing apparatus |
| CN101832884A (en) * | 2009-03-13 | 2010-09-15 | 江远仕 | Section of trigeminal nerve root and method for preparing same |
| CN102243154A (en) * | 2011-04-02 | 2011-11-16 | 中山大学附属第一医院 | Method for dyeing peripheral nerves slice |
| CN103185673A (en) * | 2011-12-30 | 2013-07-03 | 李朝林 | Integrative box type multifunctional histotome |
| CN103674677A (en) * | 2013-12-17 | 2014-03-26 | 哈尔滨医科大学 | Preparation method of brain tissue rapid freezing section |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1004904B (en) * | 1987-06-29 | 1989-07-26 | 蒋俊辉 | One step polychromatic dyeing technique for plant tissue and the specified dyestuff composite thereof |
| CN100567941C (en) * | 2005-08-15 | 2009-12-09 | 段增强 | Technique for fabricating label type biologic slice |
| US8158381B2 (en) * | 2008-03-27 | 2012-04-17 | Richard-Allan Scientific Company | Methods for integrated tissue processing and staining |
| CN101324493A (en) * | 2008-07-29 | 2008-12-17 | 孙翔宇 | Method for rapidly manufacturing large tissue specimen slice |
| CN103528872A (en) * | 2013-10-14 | 2014-01-22 | 中国农业大学 | Fluorescent staining technique of plant root tissue living body section and application of technique |
| CN106033058A (en) * | 2015-03-11 | 2016-10-19 | 李保华 | Histology observation method of rot pathogen in and out of branches of apple trees and pear trees |
| CN105300757A (en) * | 2015-09-22 | 2016-02-03 | 王善宗 | Quick and simple pathological frozen specimen manufacturing method |
| CN105910861B (en) * | 2016-04-13 | 2018-12-11 | 武汉沃亿生物有限公司 | A kind of automatic method of batch dyeing production histotomy |
-
2016
- 2016-04-13 CN CN201610228177.6A patent/CN105910861B/en active Active
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2017
- 2017-04-10 WO PCT/CN2017/079847 patent/WO2017177870A1/en active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101458182A (en) * | 2008-12-22 | 2009-06-17 | 华中科技大学 | Method for producing brain sample of small animal |
| CN101832884A (en) * | 2009-03-13 | 2010-09-15 | 江远仕 | Section of trigeminal nerve root and method for preparing same |
| CN201532333U (en) * | 2009-11-09 | 2010-07-21 | 徐华 | Histiocyte section totally-enclosed full-automatic dyeing apparatus |
| CN102243154A (en) * | 2011-04-02 | 2011-11-16 | 中山大学附属第一医院 | Method for dyeing peripheral nerves slice |
| CN103185673A (en) * | 2011-12-30 | 2013-07-03 | 李朝林 | Integrative box type multifunctional histotome |
| CN103674677A (en) * | 2013-12-17 | 2014-03-26 | 哈尔滨医科大学 | Preparation method of brain tissue rapid freezing section |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105910861A (en) | 2016-08-31 |
| WO2017177870A1 (en) | 2017-10-19 |
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Effective date of registration: 20161223 Address after: 430074 Hubei Province, East Lake Development Zone, high tech Avenue, No. 999, the future of science and Technology City, building C2, building, floor, building 1 Applicant after: Wuhan Woyi Biology Co., Ltd. Address before: 430074 Hubei Province, Wuhan city Hongshan District Luoyu Road No. 1037 Applicant before: Huazhong Science &. Technology Univ. |
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