CN105938064A - Application of reagent dose reducing histochemical pen in immunohistochemical detection of cell growing slides - Google Patents
Application of reagent dose reducing histochemical pen in immunohistochemical detection of cell growing slides Download PDFInfo
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- CN105938064A CN105938064A CN201610404344.8A CN201610404344A CN105938064A CN 105938064 A CN105938064 A CN 105938064A CN 201610404344 A CN201610404344 A CN 201610404344A CN 105938064 A CN105938064 A CN 105938064A
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- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 13
- 230000001744 histochemical effect Effects 0.000 title claims abstract description 10
- 238000013115 immunohistochemical detection Methods 0.000 title claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 230000009194 climbing Effects 0.000 claims description 23
- QGJOPFRUJISHPQ-UHFFFAOYSA-N Carbon disulfide Chemical compound S=C=S QGJOPFRUJISHPQ-UHFFFAOYSA-N 0.000 claims description 21
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 14
- 238000004043 dyeing Methods 0.000 claims description 13
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 claims description 10
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims description 10
- 239000011148 porous material Substances 0.000 claims description 9
- 239000012188 paraffin wax Substances 0.000 claims description 8
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- 241000699670 Mus sp. Species 0.000 claims description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 6
- RSWGJHLUYNHPMX-ONCXSQPRSA-N abietic acid Chemical compound C([C@@H]12)CC(C(C)C)=CC1=CC[C@@H]1[C@]2(C)CCC[C@@]1(C)C(O)=O RSWGJHLUYNHPMX-ONCXSQPRSA-N 0.000 claims description 5
- 239000007766 cera flava Substances 0.000 claims description 5
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 claims description 4
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 claims description 4
- 239000004793 Polystyrene Substances 0.000 claims description 4
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 3
- 229930040373 Paraformaldehyde Natural products 0.000 claims description 3
- 229920004890 Triton X-100 Polymers 0.000 claims description 3
- 239000013504 Triton X-100 Substances 0.000 claims description 3
- 239000012122 aqueous mounting media Substances 0.000 claims description 3
- 230000004069 differentiation Effects 0.000 claims description 3
- 238000004821 distillation Methods 0.000 claims description 3
- 239000003350 kerosene Substances 0.000 claims description 3
- 229920002866 paraformaldehyde Polymers 0.000 claims description 3
- 238000012797 qualification Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000012224 working solution Substances 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 abstract description 8
- 239000011521 glass Substances 0.000 abstract description 8
- 238000011532 immunohistochemical staining Methods 0.000 abstract description 7
- 238000009792 diffusion process Methods 0.000 abstract description 4
- 238000010186 staining Methods 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 abstract 5
- 238000004113 cell culture Methods 0.000 abstract 1
- 210000002865 immune cell Anatomy 0.000 abstract 1
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000003365 immunocytochemistry Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000003245 working effect Effects 0.000 description 1
Classifications
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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
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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
- G01N1/31—Apparatus therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- Urology & Nephrology (AREA)
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Abstract
The invention discloses application of a reagent dose reducing histochemical pen in immunohistochemical detection of cell growing slides. Specifically, cells are inoculated into a glass slide type culture dish to let cells grow on a glass slide, and immune cell histochemical staining identification is carried out at the end of cell culture. The glass slide type culture dish is provided with a culture dish body and a dish cover, the culture dish body falls into two types, i.e. a single-hole chamber and a porous chamber. The area of each hole chamber is larger than the glass slide, the dish bottom is directly used as a glass slide, the outside of the dish bottom is provided with a rectangular frame trace line dent corresponding to the glass slide size, and under the action of an external force, the dish bottom can get off along with the rectangular frame trace line dent. The histochemical pen is applicable to various immunohistochemical staining experiments of glass slides, can significantly reduce the dosage of antibodies and reagents, avoid liquid flow and diffusion during staining, and improve the operation speed, and is particularly applicable large-scale, large sample and multi-group immunohistochemical staining in the experimental study of cell growing slides or cell smears.
Description
The application is application number: 201410500701.1, the applying date: 2014.9.26, title: the divisional application of " application in cell climbing sheet Immunohistochemical detection of the multipurpose SABC pen ".
Technical field
The present invention relates to the application in cell climbing sheet Immunohistochemical detection of a kind of SABC pen.
Background technology
With specific antibody in tissue slice and cell climbing sheet thereof some chemical constituents analysis labelling and content is organized and cell in-situ is qualitative, location or quantitative study, this technology is referred to as immunocytochemistry (immunocytochemistry) technology.
Generally cell climbing sheet Immunohistochemical detection is that individual cell climbing sheet carries out dying operation one by one, typically single sample can only be done a kind of antibody, albumen and the detection of gene, it is difficult to competent extensive, the medical experiment scientific research of multisample and many groups.
Summary of the invention
It is an object of the invention to provide one and be applicable to the experiment of various immunohistochemical staining, antibody and reagent dosage can be substantially reduced, liquid trickling and diffusion when avoiding dyeing, the application in cell climbing sheet Immunohistochemical detection of the multipurpose SABC pen of raising speed of operation.
The technical solution of the present invention is:
The application in cell climbing sheet Immunohistochemical detection of a kind of multipurpose SABC pen, is characterized in that: is inoculated in by cell in microscope slide formula culture dish and carries out cell climbing sheet, and cell is cultivated after terminating and carried out immunocyte histochemical stain qualification；Described microscope slide formula culture dish is provided with culture dish body and ware lid, culture dish build number point single hole room and many pore chambers two kinds；Each pore chamber area is more than microscope slide, uses directly as microscope slide at the bottom of ware body, and lateral surface at the bottom of ware body is provided with the rectangle frame trace line chase corresponding with microscope slide size, can be deviate from along rectangle frame trace line chase the bottom of ware body under external force；
Described immunocyte histochemical stain is identified and is comprised the following steps: successively
(1) fix 15 min with ice acetone or 4% paraformaldehyde is fixed；
(2) flowing water rinsing, by PBS specimen 3 times；
(3) 10 min are hatched with 0.5%Triton X-100；
(4) 0.3%H2O2Hatch 10 min；
(5) with after PBS specimen 3 times, hot blast is the driest；
(7) with SABC pen, ink is drawn on cell climbing sheet a point 2-20 separation dyeing district；
(8) air is the driest；
(9) 10 min are hatched with normal two antiserums closings；
(10) drip mice respectively in each cell dyeing district or the anti-first antibody of rabbit hatches 30～60 min；
(11) each cell dyeing district liquid is absorbed and by PBS specimen 3 times；
(12) dropping enzyme connection mice or rabbit second antibody working solution hatch 30～60 min；
(13) PBS specimen 3 times；
(14) DAB colour developing, lucifuge, Microscopic observation；
(15) distillation washing；
(16) haematoxylin lining dye；
(17) hydrochloride alcohol differentiation, washes from the beginning；
(18) aqueous mounting medium mounting.
Described ink is prepared from the following ingredients in percentage by mixing and forms: Colophonium 2 ~ 6%, Cera Flava 18 ~ 26%, paraffin 3 ~ 8%, diisopropyl ether 5 ~ 10%, dichloromethane 5 ~ 10%, gasoline 25 ~ 30%, Carbon bisulfide 12 ~ 17%, carbon tetrachloride 2 ~ 6%, chloroform 2 ~ 6%, cyclohexanone 2 ~ 6%；Above-mentioned each amounts of components sum is 100%；
Or described ink is made up of following component mixing: polystyrene 2 ~ 4%, DMF9 ~ 11%, THF9 ~ 11%, Colophonium 2 ~ 4%, Cera Flava 13 ~ 16%, paraffin 2 ~ 4%, diisopropyl ether 4 ~ 6%, dichloromethane 4 ~ 6%, gasoline 18 ~ 22%, Carbon bisulfide 9 ~ 11%, carbon tetrachloride 9 ~ 11%, chloroform 1 ~ 2%, cyclohexanone 1 ~ 2%, kerosene 2 ~ 4%, Oleum Terebinthinae 1 ~ 2%；Above-mentioned each amounts of components sum is 100%；
Or described ink is made up of following component mixing: stearic acid 30 ~ 36%, benzene 15 ~ 20%, 46 DEG C of paraffin 9 ~ 15%, diisopropyl ether 7 ~ 11%, Carbon bisulfide 15 ~ 20%, chloroform 15 ~ 20%；Above-mentioned each amounts of components sum is 100%.
Before the bottom of ware body can being deviate from along rectangle frame trace line chase under external force, first draw with sharp keen cutter chase along the line, hook, then add in ware bottom surface and the single hole bottom land of ware body can be deviate from by external force.
Described SABC pen includes pen container, arranges spongioid cylinder pen core in pen container, is perfused with ink in pen container, and spongioid cylinder pen core front end arranges wooden pen core；Spongioid cylinder pen core is placed in pen container, and pen container is cuboid or cylinder, has screw thread to mate with the female thread of the interior cap for brush and coincide outside pen container mouth；Pen container is placed stainless shot one, when rocking pen container, makees to mix the effect of ink；The described interior cap for brush plays closing pen container mouth, prevent ink volatilization from keeping its liquid phase state and coupling the effect of the outer cap for brush, its inner surface has the female thread closed with pen container tone, can spiral cover pen container mouth, the outer surface of the interior cap for brush be provided with mate with outer cap for brush inner lip coincide fore shaft recessed, cylinder pen core end side is provided with ink brush；It is recessed that the inner lip week of the outer cap for brush is provided with fore shaft, mates with interior cap for brush outer lip and coincide, and in a closing, the cap for brush and the fixing interior cap for brush are integrated and open pen container mouth, and act the effect proposing ink brush.
One angle of many pore chambers culture dish is bevel-faced form, facilitates bearing mark；One angle of culture dish lid is in the bevel-faced form coordinated with the angle that culture dish is inclined-plane；Culture dish lid and the interleave depth >=10mm of culture dish anastomotic stoma periphery.
The standards such as the present invention is conducive to the dyeing of SABC, and staining procedure is similar to Immunohistochemistry, but the temperature of sample process, the time, reagent concentration are consistent, and an anti-kind of labelling is many, and comparability and reliability are significantly increased！The sample size processed is many, efficient quick.PBS, sera incubation, labelling two resist, and the experimental implementation that lining dye (haematoxylin) etc. need not separate can synchronize to process, and convenient and swift, experiment condition standard is consistent and easy to control.Using large area cell climbing sheet to carry out cell and cultivate its temperature processed, time, the standard such as reagent concentration is consistent, makes the comparability of experimental result and reliability be significantly increased.The quantity of Tissue Culture Dish hole slot reduces, easy to operate, efficient quick.Specific ink formulations has fully ensured that working effect.It is applicable to the experiment of various immunohistochemical staining, including；The immunohistochemical staining experiment of paraffin tissue sections, frozen tissue section and cell climbing sheet, can substantially reduce antibody and reagent dosage, it is to avoid liquid trickling and diffusion during dyeing, improves speed of operation；Be especially applicable in the experimentation of cell climbing sheet or cell smear carrying out extensive, the many groups of large sample immunohistochemical staining.
This group pen is applicable on the cell climbing sheet of tissue slice that glass is carrier and polystyrene material carrier carry out various immunohistochemical staining experiment, can substantially reduce antibody and reagent dosage, it is to avoid liquid trickling and diffusion during dyeing, improves speed of operation.Can carry out extensive, the medical experiment scientific research of multisample and many groups.
Accompanying drawing explanation
The invention will be further described with embodiment below in conjunction with the accompanying drawings.
Fig. 1 is the structural representation of culture dish of the present invention.
Fig. 2 is the structural representation of SABC pen.
Fig. 3 is abjection schematic diagram at the bottom of ware body.
Detailed description of the invention
The application in cell climbing sheet Immunohistochemical detection of a kind of multipurpose SABC pen, is inoculated in cell in microscope slide formula culture dish and carries out cell climbing sheet, and cell is cultivated after terminating and carried out immunocyte histochemical stain qualification；Described microscope slide formula culture dish is provided with culture dish body 1 and ware lid 2, culture dish build number point single hole room and many pore chambers two kinds；Each pore chamber area is more than microscope slide, and at the bottom of ware body, 3 use directly as microscope slide, and lateral surface at the bottom of ware body is provided with the rectangle frame trace line chase 4 corresponding with microscope slide size, can be deviate from along rectangle frame trace line chase the bottom of ware body under external force；
Described immunocyte histochemical stain is identified and is comprised the following steps: successively
(1) fix 15 min with ice acetone or 4% paraformaldehyde is fixed；
(2) flowing water rinsing, by PBS specimen 3 times；
(3) 10 min are hatched with 0.5%Triton X-100；
(4) 0.3%H2O2Hatch 10 min；
(5) with after PBS specimen 3 times, hot blast is the driest；
(7) with SABC pen, ink is drawn on cell climbing sheet a point 2-20 separation dyeing district；
(8) air is the driest；
(9) 10 min are hatched with normal two antiserums closings；
(10) drip mice respectively in each cell dyeing district or the anti-first antibody of rabbit hatches 30～60 min；
(11) each cell dyeing district liquid is absorbed and by PBS specimen 3 times；
(12) dropping enzyme connection mice or rabbit second antibody working solution hatch 30～60 min；
(13) PBS specimen 3 times；
(14) DAB colour developing, lucifuge, Microscopic observation；
(15) distillation washing；
(16) haematoxylin lining dye；
(17) hydrochloride alcohol differentiation, washes from the beginning；
(18) aqueous mounting medium mounting.
Described ink is prepared from the following ingredients in percentage by mixing and forms: Colophonium 2 ~ 6%, Cera Flava 18 ~ 26%, paraffin 3 ~ 8%, diisopropyl ether 5 ~ 10%, dichloromethane 5 ~ 10%, gasoline 25 ~ 30%, Carbon bisulfide 12 ~ 17%, carbon tetrachloride 2 ~ 6%, chloroform 2 ~ 6%, cyclohexanone 2 ~ 6%；Above-mentioned each amounts of components sum is 100%；
Or described ink is made up of following component mixing: polystyrene 2 ~ 4%, DMF9 ~ 11%, THF9 ~ 11%, Colophonium 2 ~ 4%, Cera Flava 13 ~ 16%, paraffin 2 ~ 4%, diisopropyl ether 4 ~ 6%, dichloromethane 4 ~ 6%, gasoline 18 ~ 22%, Carbon bisulfide 9 ~ 11%, carbon tetrachloride 9 ~ 11%, chloroform 1 ~ 2%, cyclohexanone 1 ~ 2%, kerosene 2 ~ 4%, Oleum Terebinthinae 1 ~ 2%；Above-mentioned each amounts of components sum is 100%；
Or described ink is made up of following component mixing: stearic acid 30 ~ 36%, benzene 15 ~ 20%, 46 DEG C of paraffin 9 ~ 15%, diisopropyl ether 7 ~ 11%, Carbon bisulfide 15 ~ 20%, chloroform 15 ~ 20%；Above-mentioned each amounts of components sum is 100%.
Before the bottom of ware body can being deviate from along rectangle frame trace line chase under external force, first draw with sharp keen cutter chase along the line, hook, then add in ware bottom surface and the single hole bottom land of ware body can be deviate from by external force.
Described SABC pen includes pen container 5, arranges spongioid cylinder pen core 6 in pen container, is perfused with ink in pen container, and spongioid cylinder pen core front end arranges wooden pen core 10；Spongioid cylinder pen core is placed in pen container, and pen container is cuboid or cylinder, has screw thread to mate with the female thread of the interior cap for brush 7 and coincide outside pen container mouth；Pen container is placed stainless shot 8 one, when rocking pen container, makees to mix the effect of ink；The described interior cap for brush plays closing pen container mouth, prevent ink volatilization from keeping its liquid phase state and coupling the effect of the outer cap for brush, its inner surface has the female thread closed with pen container tone, can spiral cover pen container mouth, the outer surface of the interior cap for brush be provided with mate with the outer cap for brush 9 inner lip coincide fore shaft recessed, cylinder pen core end side is provided with ink brush 11；It is recessed that the inner lip week of the outer cap for brush is provided with fore shaft, mates with interior cap for brush outer lip and coincide, and in a closing, the cap for brush and the fixing interior cap for brush are integrated and open pen container mouth, and act the effect proposing ink brush.Ink exchange hole 12 is set on the interior cap for brush.
One angle of many pore chambers culture dish is bevel-faced form, facilitates bearing mark；One angle of culture dish lid is in the bevel-faced form coordinated with the angle that culture dish is inclined-plane；Culture dish lid and the interleave depth >=10mm of culture dish anastomotic stoma periphery.
Claims (4)
1. the group pen reducing reagent dosage is applied in cell climbing sheet SABC detects, and it is characterized in that: be inoculated in by cell in microscope slide formula culture dish and carry out cell climbing sheet, and cell is cultivated after terminating and carried out immunocyte histochemical stain qualification；Described microscope slide formula culture dish is provided with culture dish body and ware lid, culture dish build number point single hole room and many pore chambers two kinds；Each pore chamber area is more than microscope slide, use directly as microscope slide at the bottom of ware body, lateral surface at the bottom of ware body is provided with the rectangle frame trace line chase corresponding with microscope slide size, the bottom of ware body can be deviate from along rectangle frame trace line chase under external force, obtain and have cultivation cell face just as at the bottom of the ware of microscope slide, be ready for use on various Immunohistochemical detection；
Described immunocyte histochemical stain is identified and is comprised the following steps: successively
(1) fix 15 min with ice acetone or 4% paraformaldehyde is fixed；
(2) flowing water rinsing, by PBS specimen 3 times；
(3) 10 min are hatched with 0.5%Triton X-100；
(4) 0.3%H2O2Hatch 10 min；
(5) with after PBS specimen 3 times, hot blast is the driest；
(7) with SABC pen, ink is drawn on cell climbing sheet a point 2-20 separation dyeing district；
(8) air is the driest；
(9) 10 min are hatched with normal two antiserums closings；
(10) drip mice respectively in each cell dyeing district or the anti-first antibody of rabbit hatches 30～60 min；
(11) each cell dyeing district liquid is absorbed and by PBS specimen 3 times；
(12) dropping enzyme connection mice or rabbit second antibody working solution hatch 30～60 min；
(13) PBS specimen 3 times；
(14) DAB colour developing, lucifuge, Microscopic observation；
(15) distillation washing；
(16) haematoxylin lining dye；
(17) hydrochloride alcohol differentiation, washes from the beginning；
(18) aqueous mounting medium mounting；
Described ink is made up of following component mixing: polystyrene 2 ~ 4%, DMF9 ~ 11%, THF9 ~ 11%, Colophonium 2 ~ 4%, Cera Flava 13 ~ 16%, paraffin 2 ~ 4%, diisopropyl ether 4 ~ 6%, dichloromethane 4 ~ 6%, gasoline 18 ~ 22%, Carbon bisulfide 9 ~ 11%, carbon tetrachloride 9 ~ 11%, chloroform 1 ~ 2%, cyclohexanone 1 ~ 2%, kerosene 2 ~ 4%, Oleum Terebinthinae 1 ~ 2%；Above-mentioned each amounts of components sum is 100%.
The group pen of minimizing reagent dosage the most according to claim 1 is applied in cell climbing sheet SABC detects, it is characterized in that: before the bottom of ware body can being deviate from along rectangle frame trace line chase under external force, first draw with sharp keen cutter chase along the line, hook, then the single hole bottom land of ware body can be deviate from by external force a little in ware bottom surface.
The group pen of minimizing reagent dosage the most according to claim 1 and 2 is applied in cell climbing sheet SABC detects, it is characterized in that: described SABC pen includes pen container, spongioid cylinder pen core is set in pen container, being perfused with ink in pen container, spongioid cylinder pen core front end arranges wooden pen core.
The group pen of minimizing reagent dosage the most according to claim 1 and 2 is applied in cell climbing sheet SABC detects, and it is characterized in that: an angle of many pore chambers culture dish is bevel-faced form, facilitates bearing mark；One angle of culture dish lid is in the bevel-faced form coordinated with the angle that culture dish is inclined-plane；Culture dish lid and the interleave depth >=10mm of culture dish anastomotic stoma periphery.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610404344.8A CN105938064B (en) | 2014-09-26 | 2014-09-26 | The group pen for reducing reagent dosage is applied in the detection of cell climbing sheet immunohistochemistry |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410500701.1A CN104359741B (en) | 2014-09-26 | 2014-09-26 | Application of multi-functional PAP pen in cell climbing immunohistochemical detection |
| CN201610404344.8A CN105938064B (en) | 2014-09-26 | 2014-09-26 | The group pen for reducing reagent dosage is applied in the detection of cell climbing sheet immunohistochemistry |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410500701.1A Division CN104359741B (en) | 2014-09-26 | 2014-09-26 | Application of multi-functional PAP pen in cell climbing immunohistochemical detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105938064A true CN105938064A (en) | 2016-09-14 |
| CN105938064B CN105938064B (en) | 2019-03-19 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610404343.3A Active CN106092703B (en) | 2014-09-26 | 2014-09-26 | Application of the easy to operate groupization pen in the detection of cell climbing sheet immunohistochemistry |
| CN201610404342.9A Active CN106092702B (en) | 2014-09-26 | 2014-09-26 | The group pen for reducing reagent dosage is applied in the detection of cell climbing sheet immunohistochemistry |
| CN201610404345.2A Active CN105938065B (en) | 2014-09-26 | 2014-09-26 | The fast group pen of service speed is applied in the detection of cell climbing sheet immunohistochemistry |
| CN201410500701.1A Active CN104359741B (en) | 2014-09-26 | 2014-09-26 | Application of multi-functional PAP pen in cell climbing immunohistochemical detection |
| CN201610404344.8A Active CN105938064B (en) | 2014-09-26 | 2014-09-26 | The group pen for reducing reagent dosage is applied in the detection of cell climbing sheet immunohistochemistry |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201610404343.3A Active CN106092703B (en) | 2014-09-26 | 2014-09-26 | Application of the easy to operate groupization pen in the detection of cell climbing sheet immunohistochemistry |
| CN201610404342.9A Active CN106092702B (en) | 2014-09-26 | 2014-09-26 | The group pen for reducing reagent dosage is applied in the detection of cell climbing sheet immunohistochemistry |
| CN201610404345.2A Active CN105938065B (en) | 2014-09-26 | 2014-09-26 | The fast group pen of service speed is applied in the detection of cell climbing sheet immunohistochemistry |
| CN201410500701.1A Active CN104359741B (en) | 2014-09-26 | 2014-09-26 | Application of multi-functional PAP pen in cell climbing immunohistochemical detection |
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| CN105865879A (en) * | 2016-05-19 | 2016-08-17 | 四川金域医学检验中心有限公司 | Operation method for immunohistochemical staining of frozen sections |
| CN106248464B (en) * | 2016-09-02 | 2020-06-09 | 四川大学 | Cell climbing sheet efficient dyeing device |
| CN107699486A (en) * | 2017-09-22 | 2018-02-16 | 山东省农业科学院畜牧兽医研究所 | For dyeing the Tissue Culture Dish and cell culture dyeing, observational technique of observation |
| CN109100503B (en) * | 2018-09-20 | 2021-02-02 | 同济大学 | Immunohistochemical pen liquid and preparation method thereof |
| CN110887720A (en) * | 2019-12-11 | 2020-03-17 | 武汉原谷生物科技有限责任公司 | Immunohistochemical pen semisolid pen paste and preparation method thereof |
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| CN106092703A (en) | 2016-11-09 |
| CN106092703B (en) | 2018-07-17 |
| CN106092702A (en) | 2016-11-09 |
| CN105938064B (en) | 2019-03-19 |
| CN104359741B (en) | 2017-01-11 |
| CN106092702B (en) | 2018-07-06 |
| CN104359741A (en) | 2015-02-18 |
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| CN105938065B (en) | 2019-03-12 |
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