CN105910861A - Automatic method for making tissue slices through batch dyeing - Google Patents
Automatic method for making tissue slices through batch dyeing Download PDFInfo
- Publication number
- CN105910861A CN105910861A CN201610228177.6A CN201610228177A CN105910861A CN 105910861 A CN105910861 A CN 105910861A CN 201610228177 A CN201610228177 A CN 201610228177A CN 105910861 A CN105910861 A CN 105910861A
- Authority
- CN
- China
- Prior art keywords
- sample
- layer part
- dye
- slices
- dyeing
- 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
- 238000004043 dyeing Methods 0.000 title claims abstract description 17
- 238000000034 method Methods 0.000 title abstract description 20
- 210000004556 brain Anatomy 0.000 claims abstract description 18
- 230000035515 penetration Effects 0.000 claims abstract description 4
- 239000000975 dye Substances 0.000 claims description 16
- 239000012530 fluid Substances 0.000 claims description 9
- 239000012466 permeate Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 abstract 3
- 238000003384 imaging method Methods 0.000 description 3
- 238000006424 Flood reaction Methods 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009940 knitting Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 238000004064 recycling Methods 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
- G01N1/286—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
- G01N1/31—Apparatus therefor
-
- 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
- G01N1/312—Apparatus therefor for samples mounted on planar substrates
-
- 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
Landscapes
- 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 an automatic method for making tissue slices through batch dyeing. The method comprises the following steps: 1, immersing a sample to be processed in a processing liquid containing a cytoarchitectonic dye; 2, allowing dye molecules in the processing liquid to penetrate to the depth of the sample from the surface of the sample in order to dye cytoarchitecture at the shallow layer part of the surface of the sample after a period of time; 3, slicing the dyed shallow layer part of the sample by using a cutter; 4, allowing the dye molecules in the processing liquid to penetrate to the shallow layer part cytoarchitecture from the sections of slices in order to dye the shallow layer part cytoarchitecture, and carrying out computer control to lift the sample to be in a same height with the thickness of the slices; and 5, repeating step 3 to step 4 until the whole sample is cut to form cytoarchitecture dyed brain slices. The method avoids manual operation interference and saves time and labor through utilizing natural penetration of the dye in slice gaps in the dyeing process, and is especially suitable for batch dyeing and making tissue slices.
Description
Technical field
The present invention relates to the making of biological tissue section, make tissue in particular to one batch dyeing and cut
The automatic mode of sheet, belongs to life science technical field.
Background technology
In order to study neuromechanism and the function information of Different brain region in Jie's sight or microscopic scale, researcher leads to
Often needing to be cut into cerebral tissue the brain sheet of 100 microns or thinner, recycling labelling and imaging means are to this
A little brain sheets carry out dyeing piecewise, imaging.
Owing to nervous system exists larger difference with brain Division, the nerve of Different brain region, therefore, right
When neuromechanism and function information are analyzed, need first its brain district, place to be positioned.At present,
The diversity that is distributed mainly by Different brain region cytoarchitectonic of method realizing location, brain district is carried out,
After brain sheet is carried out cytoarchitectonic dyeing, it is possible to obtain the cytoarchitectonic information of this brain sheet, control reference
Brain map, such as " Allen Reference Atlas ", the similar imaging results of comparison, to confirm brain district, place.
But, such acquisition of information, it is common that carry out the manual process dyeed after experience section the most piecewise,
Even if Mouse brain outer profile size also reaches more than 10 millimeters, need to be cut into tens or hundreds of brain sheets.Raw
Thing experiment generally requires and carries out great many of experiments to obtain statistically rule, eliminates individual difference and brings
Error.Therefore, this poor efficiency is laborious section, colouring method can not meet the needs of existing research.
The method urgently develop the batch dyeing of a kind of automatization, making tissue slice.
Summary of the invention
Present invention aim to overcome that above-mentioned the deficiencies in the prior art provide a kind of batch dyeing making group
Knitting the automatic mode of section, the method comprises the following steps:
(1) sample to be processed is immersed in the working fluid containing cytoarchitectonic dyestuff；
(2) dye molecule in working fluid permeates to sample depths at sample surface, through a timing
After between, the shallow-layer part cytoarchitectonic of sample surface is colored；
(3) utilize cutter that the shallow-layer part that sample is dyed is cut into slices；
(4) to infiltrate into shallow-layer part to sample depths at slice profile thin for the dye molecule in working fluid
Born of the same parents construct and are colored, and will be controlled by computer, by height identical with slice thickness for sample lifting；
(5) repeat the above steps (3)～(4), until being cut into cytoarchitectonic dyeing by whole sample
Brain sheet.
In technique scheme, described sample side scribbles the medium preventing dyestuff penetration.
The inventive method has the advantage that
Dyeing being carried out with slicing processes automatization, the operation of the inventive method is first to sample surface
Shallow-layer part cytoarchitectonic dyes, and then cuts into slices the shallow-layer part that sample is dyed, and circulates
The operation of above-mentioned " dyeing-section ", it is thus possible to realize the inventive method by the way of automatically controlling.
The method, compared with original working method first cut into slices and dye the most piecewise, improves and carries out brain sheet piecewise
The efficiency processed.Dyeing course utilizes the naturally osmotic at section gap dyestuff, it is to avoid manual operations
Interference, time saving and energy saving.
Present invention is especially suited for batch to dye, make tissue slice, for obtaining Neuroscience Research on a large scale
Experimental data provide new experimental tool.
Accompanying drawing explanation
Fig. 1 is the schematic diagram being made tissue slice by the inventive method batch dyeing.
Detailed description of the invention
The present invention is described in further detail with specific embodiment below in conjunction with the accompanying drawings.
As it is shown in figure 1, sample 1 is first fixed in tank 3 by the inventive method, tank 3 is full of
Working fluid 2 containing cytoarchitectonic dyestuff, floods completely by sample 1.The side of sample used by the present invention
Topcoating has the medium preventing dyestuff penetration, such as nial polish so that dyestuff cannot ooze from the side of sample
Thoroughly.Standing after enough time, working fluid 2 permeates in sample 1 so that surface shallow-layer sample thin
Born of the same parents construct and are colored.Utilize cutter 5 that sample 1 is cut into slices so that dyed shallow-layer part 4
Cut off sample, exposed next to be dyed, the surface of section.Dye molecule in working fluid is from cutting
Sheet section part infiltrates into shallow-layer part cytoarchitectonic and is colored to sample depths, is automatically controlled by computer
System, by height identical with slice thickness for sample lifting.The circulation of dyeing-slicing processes can automatization repeat
Carry out, until sample is cut.
Claims (2)
1. the automatic mode of a batch dyeing making tissue slice, it is characterised in that including:
(1) sample to be processed is immersed in the working fluid containing cytoarchitectonic dyestuff；
(2) dye molecule in working fluid permeates to sample depths at sample surface, through a timing
After between, the shallow-layer part cytoarchitectonic of sample surface is colored；
(3) utilize cutter that the shallow-layer part that sample is dyed is cut into slices；
(4) to infiltrate into shallow-layer part to sample depths at slice profile thin for the dye molecule in working fluid
Born of the same parents construct and are colored, and will be controlled by computer, by height identical with slice thickness for sample lifting；
(5) repeat the above steps (3)～(4), until being cut into cytoarchitectonic dyeing by whole sample
Brain sheet.
The most according to claim 1, batch dyeing makes the automatic mode of tissue slice, and it is special
Levy and be: described sample side scribbles the medium 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 true CN105910861A (en) | 2016-08-31 |
| CN105910861B CN105910861B (en) | 2018-12-11 |
Family
ID=56746967
Family Applications (1)
| 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) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017177870A1 (en) * | 2016-04-13 | 2017-10-19 | 武汉沃亿生物有限公司 | Automated batch method for staining and making tissue slices |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1916590A (en) * | 2005-08-15 | 2007-02-21 | 段增强 | Technique for fabricating label type biologic slice |
| CN101324493A (en) * | 2008-07-29 | 2008-12-17 | 孙翔宇 | Method for rapidly manufacturing large tissue specimen slice |
| 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 |
| CN105300757A (en) * | 2015-09-22 | 2016-02-03 | 王善宗 | Quick and simple pathological frozen specimen manufacturing method |
Family Cites Families (5)
| 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 |
| US8158381B2 (en) * | 2008-03-27 | 2012-04-17 | Richard-Allan Scientific Company | Methods for integrated tissue processing and staining |
| 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 |
| 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
-
2017
- 2017-04-10 WO PCT/CN2017/079847 patent/WO2017177870A1/en active Application Filing
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1916590A (en) * | 2005-08-15 | 2007-02-21 | 段增强 | Technique for fabricating label type biologic slice |
| CN101324493A (en) * | 2008-07-29 | 2008-12-17 | 孙翔宇 | Method for rapidly manufacturing large tissue specimen slice |
| 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 |
| CN105300757A (en) * | 2015-09-22 | 2016-02-03 | 王善宗 | Quick and simple pathological frozen specimen manufacturing method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017177870A1 (en) * | 2016-04-13 | 2017-10-19 | 武汉沃亿生物有限公司 | Automated batch method for staining and making tissue slices |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2017177870A1 (en) | 2017-10-19 |
| CN105910861B (en) | 2018-12-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105910861A (en) | Automatic method for making tissue slices through batch dyeing | |
| DE112010002246T5 (en) | DNA analyzer | |
| CA3034310A1 (en) | Methods and apparatus for manufacturing a microfluidic arrangement, and a microfluidic arrangement | |
| EP3173490A3 (en) | Cell selection method, cell detection method, cell selection apparatus, and cell detection apparatus | |
| CN109975090B (en) | Preparation method of thyroid and mammary gland fine needle punctured cell tissue block | |
| DE4409436A1 (en) | Process for processing nucleic acids | |
| CN106102872A (en) | Electrophoretic techniques is used to extract from unicellular synchronization and separate RNA and DNA | |
| CN106447644A (en) | New method for quantitative analysis of microscopic oil displacement experiment | |
| CN104764645A (en) | Amphibian phalanx paraffin section manufacturing method | |
| CN103471898A (en) | Rapid neuron staining method based on Golgi silver staining method | |
| Wang et al. | Use of threshold flux concept to aid selection of sustainable operating flux: A multi-scale study from laboratory to full scale | |
| CN211602632U (en) | Cell chip | |
| CN110423673B (en) | In-situ culture device and method for separating and screening microorganisms at high flux | |
| CN205368351U (en) | Single cellular localization microporous membrane and unicellular automatic acquisition device | |
| CN102175880B (en) | Sampling height adjuster of porous plate | |
| CN105891208B (en) | For constructing the architecture cytoarchitectonic information acquisition method of standard diagram data set | |
| CN105259001B (en) | A kind of paraffin section pre-treating method for tissue biopsy | |
| Masri | Uncovering the potential of ultra scale-down tools to enable cost-effective “Quality by Design” | |
| CN106029234B (en) | Porous membrane patterning techniques | |
| CN204249018U (en) | A kind of drawer's panel puncher | |
| DE102017130395A1 (en) | Method for spatially resolved, fluid-assisted examination of a microscopic sample, compartmentation device and examination system arranged on a microscope slide | |
| Lopez Pena | Computational derivation of conditions for upscalability of bioclogging in pore network models | |
| CN206399632U (en) | A kind of optical wave plate optical axis labelling apparatus | |
| CN207983449U (en) | A kind of pcb board trimming device | |
| DE3237831C2 (en) | Device for the analysis of proteins by electrophoresis and photometry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
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. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |