CN104359741A - Application of multi-functional PAP pen in cell climbing immunohistochemical detection - Google Patents
Application of multi-functional PAP pen in cell climbing immunohistochemical detection Download PDFInfo
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- CN104359741A CN104359741A CN201410500701.1A CN201410500701A CN104359741A CN 104359741 A CN104359741 A CN 104359741A CN 201410500701 A CN201410500701 A CN 201410500701A CN 104359741 A CN104359741 A CN 104359741A
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- 230000009194 climbing Effects 0.000 title claims abstract description 24
- 238000013115 immunohistochemical detection Methods 0.000 title claims abstract description 13
- 238000004043 dyeing Methods 0.000 claims abstract description 17
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 239000011148 porous material Substances 0.000 claims abstract description 5
- QGJOPFRUJISHPQ-UHFFFAOYSA-N Carbon disulfide Chemical compound S=C=S QGJOPFRUJISHPQ-UHFFFAOYSA-N 0.000 claims description 18
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 18
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 18
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 claims description 12
- 239000012188 paraffin wax Substances 0.000 claims description 10
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 9
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 claims description 9
- RSWGJHLUYNHPMX-UHFFFAOYSA-N Abietic-Saeure Natural products C12CCC(C(C)C)=CC2=CCC2C1(C)CCCC2(C)C(O)=O RSWGJHLUYNHPMX-UHFFFAOYSA-N 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 6
- KHPCPRHQVVSZAH-HUOMCSJISA-N Rosin Natural products O(C/C=C/c1ccccc1)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 KHPCPRHQVVSZAH-HUOMCSJISA-N 0.000 claims description 6
- 235000013871 bee wax Nutrition 0.000 claims description 6
- 239000012166 beeswax Substances 0.000 claims description 6
- 230000000694 effects Effects 0.000 claims description 6
- 230000001744 histochemical effect Effects 0.000 claims description 6
- 238000012797 qualification Methods 0.000 claims description 6
- KHPCPRHQVVSZAH-UHFFFAOYSA-N trans-cinnamyl beta-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OCC=CC1=CC=CC=C1 KHPCPRHQVVSZAH-UHFFFAOYSA-N 0.000 claims description 6
- 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
- 238000004140 cleaning Methods 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
- 235000021355 Stearic acid Nutrition 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
- 239000012530 fluid Substances 0.000 claims description 3
- 239000003350 kerosene Substances 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 3
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 3
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 3
- 229920002866 paraformaldehyde Polymers 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 239000008117 stearic acid Substances 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
- 210000004027 cell Anatomy 0.000 claims 10
- 230000008878 coupling Effects 0.000 claims 1
- 238000010168 coupling process Methods 0.000 claims 1
- 238000005859 coupling reaction Methods 0.000 claims 1
- 210000004748 cultured cell Anatomy 0.000 claims 1
- 239000011521 glass Substances 0.000 abstract description 9
- 238000002474 experimental method Methods 0.000 abstract description 8
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 6
- 238000009792 diffusion process Methods 0.000 abstract description 4
- 230000002055 immunohistochemical effect Effects 0.000 abstract 3
- 238000004113 cell culture Methods 0.000 abstract 1
- 238000011532 immunohistochemical staining Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000003365 immunocytochemistry Methods 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
- 238000010186 staining Methods 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
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- 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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Abstract
The invention discloses an application of a multi-functional PAP pen in cell climbing immunohistochemical detection. The application comprises the following steps: inoculating cells in a glass slide-type culture dish for enabling cells to grow on the glass slide-type culture dish; after cell culture, conducting immunohistochemical dyeing identification. The glass slide-type culture dish is provided with a culture dish body and a culture dish cover, wherein the culture dish body comprises a single-pore chamber model and a porous chamber model; the area of each pore chamber is greater than that of the glass slide; the bottom of the culture dish body can be directly used as the glass slide; a rectangular streak line groove, the size of which is corresponding to that of the glass slide, is formed in the outer side surfaces of the bottom of the culture dish body; the bottom of the culture dish body can be pulled out of the rectangular streak line groove under the action of external force. The application is suitable for various immunohistochemical dyeing experiments on a glass slice, can remarkably reduce antibodies and the reagent dosage, avoids liquid flow and diffusion in dyeing, improves the operation speed, and is particularly suitable for large-scale large-sample and multi-group immunohistochemical dyeing in experimental study of cell growing on the glass slide-type culture dish or cell smearing.
Description
Technical field
The present invention relates to the application of a kind of SABC pen in cell climbing sheet Immunohistochemical detection.
Background technology
With specific antibody to the mark of some chemical constituents analysis in histotomy and cell climbing sheet thereof and content is organized and cell in-situ is qualitative, location or quantitative examination, this technology is called immunocytochemistry (immunocytochemistry) technology.
Usual cell climbing sheet Immunohistochemical detection carries out dying operation one by one to individual cell climbing sheet, generally can only do a kind of antibody to single sample, the detection of albumen and gene, is difficult to competent extensive, the medical experiment scientific research of multisample and many groups.
Summary of the invention
One is the object of the present invention is to provide to be applicable to the experiment of various immunohistochemical staining, can significantly reduce antibody and reagent dosage, when avoiding dyeing, liquid trickling and diffusion, improve the application of multi-usage SABC pen in cell climbing sheet Immunohistochemical detection of operating speed.
Technical solution of the present invention is:
The application of multi-usage SABC pen in cell climbing sheet Immunohistochemical detection, is characterized in that: be inoculated in by cell in microslide formula double dish and carry out cell climbing sheet, carries out immunocyte histochemical stain qualification after cell chulture terminates; Described microslide formula double dish is provided with double dish body and ware lid, double dish build number point single hole room and two kinds, porous room; Each pore chamber area is greater than microslide, and directly use as microslide 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 microslide size, the bottom of ware body can be deviate from along rectangle frame trace line chase under external force;
Described immunocyte histochemical stain qualification comprises the following steps: successively
(1) 15 min are fixed with ice acetone or 4% paraformaldehyde is fixed;
(2) flowing water rinsing, cleans sample 3 times with PBS;
(3) 10 min are hatched with 0.5%Triton X-100;
(4) 0.3%H
2o
2hatch 10 min;
(5) with after PBS cleaning sample 3 times, hot blast is fully dry;
(7) ink is drawn on cell climbing sheet a point 2-20 separation dyeing district with SABC pen;
(8) air is fully dry;
(9) 10 min are hatched with normal two antiserums are closed;
(10) mouse is dripped respectively in each cell dyeing district or the anti-first antibody of rabbit hatches 30 ~ 60 min;
(11) absorb each cell dyeing district liquid and clean sample 3 times with PBS;
(12) dropping enzyme connection mouse or rabbit second antibody working fluid hatch 30 ~ 60 min;
(13) PBS cleans sample 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 mixed by the component of following weight percents and forms: rosin 2 ~ 6%, beeswax 18 ~ 26%, paraffin 3 ~ 8%, isopropyl ether 5 ~ 10%, methylene chloride 5 ~ 10%, gasoline 25 ~ 30%, carbon disulphide 12 ~ 17%, phenixin 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%, rosin 2 ~ 4%, beeswax 13 ~ 16%, paraffin 2 ~ 4%, isopropyl ether 4 ~ 6%, methylene chloride 4 ~ 6%, gasoline 18 ~ 22%, carbon disulphide 9 ~ 11%, phenixin 9 ~ 11%, chloroform 1 ~ 2%, cyclohexanone 1 ~ 2%, kerosene 2 ~ 4%, terebinthina 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%, isopropyl ether 7 ~ 11%, carbon disulphide 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 comprises pen container, arranges spongioid right cylinder pen core, be perfused with ink in pen container in pen container, and spongioid right cylinder pen core front end arranges wooden pen core; Spongioid right cylinder pen core is placed in pen container, and pen container is rectangular parallelepiped or right cylinder, has screw thread to mate with the internal thread of the interior cap for brush and coincide outside pen container mouth; Place stainless shot one in pen container, when rocking pen container, do the effect of mixing ink; The described interior cap for brush plays closed pen container mouth, prevent ink from volatilizing and keep its liquid phase state and the effect connecting the outer cap for brush, its inside surface has the internal thread closed with pen container tone, can spiral cover pen container mouth, the outside surface of the interior cap for brush is provided with that to mate the fore shaft coincide with outer cap for brush inner lip recessed, and right cylinder pen core end side is provided with ink brush; The outer cap for brush inner lip week to be provided with fore shaft recessed, mate with interior cap for brush outer lip and coincide, to work in closing a cap for brush and fixing in the cap for brush be integrated and open pen container mouth, and play proposition ink brush.
An angle of porous room double dish is bevel-faced form, facilitates bearing mark; The bevel-faced form that the angle that an angle of double dish lid is is inclined-plane with double dish coordinates; Interleave depth >=the 10mm of double dish lid and double dish previous anastomotic periphery.
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, the standards such as reagent concentration are consistent, and the primary antibodie kind of mark is how, and comparability and reliability are significantly increased! The sample size of process is many, efficient quick.PBS cleaning, sera incubation, mark two resist, and lining dye (haematoxylin) etc. does not need the experimental implementation separated synchronously to process, and convenient and swift, experiment condition standard is consistent and easy to control.Adopt large area cell climbing sheet to carry out the temperature of its process of cell chulture, the time, the standards such as reagent concentration are consistent, and the comparability of experimental result and reliability are significantly increased.The quantity of Tissue Culture Dish hole slot reduces, easy to operate, efficient quick.Specific ink formulations fully ensure that working effect.Be applicable to the experiment of various immunohistochemical staining, comprise; Paraffin tissue sections, the immunohistochemical staining experiment of frozen tissue section and cell climbing sheet, can significantly reduce antibody and reagent dosage, and when avoiding dyeing, liquid trickling and diffusion, improve operating speed; Especially be applicable in the experimental study of cell climbing sheet or cell smear carry out extensive, the many groups of large sample immunohistochemical staining.
The cell climbing sheet that this group pen is applicable to histotomy that glass is carrier and polystyrene material carrier carries out various immunohistochemical staining experiment, can significantly reduce antibody and reagent dosage, when avoiding dyeing, liquid trickling and diffusion, improve operating speed.Can carry out extensive, the medical experiment scientific research of multisample and many groups.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the invention will be further described.
Fig. 1 is the structural representation of double dish of the present invention.
Fig. 2 is the structural representation of SABC pen.
Fig. 3 deviates from schematic diagram at the bottom of ware body.
Embodiment
The application of multi-usage SABC pen in cell climbing sheet Immunohistochemical detection, is inoculated in cell in microslide formula double dish and carries out cell climbing sheet, carries out immunocyte histochemical stain qualification after cell chulture terminates; Described microslide formula double dish is provided with double dish body 1 and ware lid 2, double dish build number point single hole room and two kinds, porous room; Each pore chamber area is greater than microslide, and at the bottom of ware body, 3 directly use as microslide, and lateral surface at the bottom of ware body is provided with the rectangle frame trace line chase 4 corresponding with microslide size, the bottom of ware body can be deviate from along rectangle frame trace line chase under external force;
Described immunocyte histochemical stain qualification comprises the following steps: successively
(1) 15 min are fixed with ice acetone or 4% paraformaldehyde is fixed;
(2) flowing water rinsing, cleans sample 3 times with PBS;
(3) 10 min are hatched with 0.5%Triton X-100;
(4) 0.3%H
2o
2hatch 10 min;
(5) with after PBS cleaning sample 3 times, hot blast is fully dry;
(7) ink is drawn on cell climbing sheet a point 2-20 separation dyeing district with SABC pen;
(8) air is fully dry;
(9) 10 min are hatched with normal two antiserums are closed;
(10) mouse is dripped respectively in each cell dyeing district or the anti-first antibody of rabbit hatches 30 ~ 60 min;
(11) absorb each cell dyeing district liquid and clean sample 3 times with PBS;
(12) dropping enzyme connection mouse or rabbit second antibody working fluid hatch 30 ~ 60 min;
(13) PBS cleans sample 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 mixed by the component of following weight percents and forms: rosin 2 ~ 6%, beeswax 18 ~ 26%, paraffin 3 ~ 8%, isopropyl ether 5 ~ 10%, methylene chloride 5 ~ 10%, gasoline 25 ~ 30%, carbon disulphide 12 ~ 17%, phenixin 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%, rosin 2 ~ 4%, beeswax 13 ~ 16%, paraffin 2 ~ 4%, isopropyl ether 4 ~ 6%, methylene chloride 4 ~ 6%, gasoline 18 ~ 22%, carbon disulphide 9 ~ 11%, phenixin 9 ~ 11%, chloroform 1 ~ 2%, cyclohexanone 1 ~ 2%, kerosene 2 ~ 4%, terebinthina 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%, isopropyl ether 7 ~ 11%, carbon disulphide 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 comprises pen container 5, arranges spongioid right cylinder pen core 6, be perfused with ink in pen container in pen container, and spongioid right cylinder pen core front end arranges wooden pen core 10; Spongioid right cylinder pen core is placed in pen container, and pen container is rectangular parallelepiped or right cylinder, has screw thread to mate with the internal thread of the interior cap for brush 7 and coincide outside pen container mouth; Place stainless shot 8 one in pen container, when rocking pen container, do the effect of mixing ink; The described interior cap for brush plays closed pen container mouth, prevent ink from volatilizing and keep its liquid phase state and the effect connecting the outer cap for brush, its inside surface has the internal thread closed with pen container tone, can spiral cover pen container mouth, the outside surface of the interior cap for brush is provided with that to mate the fore shaft coincide with the outer cap for brush 9 inner lip recessed, and right cylinder pen core end side is provided with ink brush 11; The outer cap for brush inner lip week to be provided with fore shaft recessed, mate with interior cap for brush outer lip and coincide, to work in closing a cap for brush and fixing in the cap for brush be integrated and open pen container mouth, and play proposition ink brush.The interior cap for brush is arranged ink and exchange hole 12.
An angle of porous room double dish is bevel-faced form, facilitates bearing mark; The bevel-faced form that the angle that an angle of double dish lid is is inclined-plane with double dish coordinates; Interleave depth >=the 10mm of double dish lid and double dish previous anastomotic periphery.
Claims (4)
1. the application of multi-usage SABC pen in cell climbing sheet Immunohistochemical detection, is characterized in that: be inoculated in by cell in microslide formula double dish and carry out cell climbing sheet, carries out immunocyte histochemical stain qualification after cell chulture terminates; Described microslide formula double dish is provided with double dish body and ware lid, double dish build number point single hole room and two kinds, porous room; Each pore chamber area is greater than microslide, directly use as microslide 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 microslide size, the bottom of ware body can be deviate from along rectangle frame trace line chase under external force, obtain and have cultured cell face just as at the bottom of the ware of microslide, be ready for use on various Immunohistochemical detection;
Described immunocyte histochemical stain qualification comprises the following steps: successively
(1) 15 min are fixed with ice acetone or 4% paraformaldehyde is fixed;
(2) flowing water rinsing, cleans sample 3 times with PBS;
(3) 10 min are hatched with 0.5%Triton X-100;
(4) 0.3%H
2o
2hatch 10 min;
(5) with after PBS cleaning sample 3 times, hot blast is fully dry;
(7) ink is drawn on cell climbing sheet a point 2-20 separation dyeing district with SABC pen;
(8) air is fully dry;
(9) 10 min are hatched with normal two antiserums are closed;
(10) mouse is dripped respectively in each cell dyeing district or the anti-first antibody of rabbit hatches 30 ~ 60 min;
(11) absorb each cell dyeing district liquid and clean sample 3 times with PBS;
(12) dropping enzyme connection mouse or rabbit second antibody working fluid hatch 30 ~ 60 min;
(13) PBS cleans sample 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.
2. the application of multi-usage SABC pen according to claim 1 in cell climbing sheet Immunohistochemical detection, it is characterized in that: described ink is mixed by the component of following weight percents and forms: rosin 2 ~ 6%, beeswax 18 ~ 26%, paraffin 3 ~ 8%, isopropyl ether 5 ~ 10%, methylene chloride 5 ~ 10%, gasoline 25 ~ 30%, carbon disulphide 12 ~ 17%, phenixin 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%, rosin 2 ~ 4%, beeswax 13 ~ 16%, paraffin 2 ~ 4%, isopropyl ether 4 ~ 6%, methylene chloride 4 ~ 6%, gasoline 18 ~ 22%, carbon disulphide 9 ~ 11%, phenixin 9 ~ 11%, chloroform 1 ~ 2%, cyclohexanone 1 ~ 2%, kerosene 2 ~ 4%, terebinthina 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%, isopropyl ether 7 ~ 11%, carbon disulphide 15 ~ 20%, chloroform 15 ~ 20%; Above-mentioned each amounts of components sum is 100%.
3. the application of multi-usage SABC pen according to claim 1 and 2 in cell climbing sheet Immunohistochemical detection, 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.
4. the application of SABC pen according to claim 1 and 2 in cell climbing sheet Immunohistochemical detection, it is characterized in that: described SABC pen comprises pen container, spongioid right cylinder pen core is set in pen container, be perfused with ink in pen container, spongioid right cylinder pen core front end arranges wooden pen core; Spongioid right cylinder pen core is placed in pen container, and pen container is rectangular parallelepiped or right cylinder, has screw thread to mate with the internal thread of the interior cap for brush and coincide outside pen container mouth; Place stainless shot one in pen container, when rocking pen container, do the effect of mixing ink; The described interior cap for brush plays closed pen container mouth, prevent ink from volatilizing and keep its liquid phase state and the effect connecting the outer cap for brush, its inside surface has the internal thread closed with pen container tone, can spiral cover pen container mouth, the outside surface of the interior cap for brush is provided with that to mate the fore shaft coincide with outer cap for brush inner lip recessed, and right cylinder pen core end side is provided with ink brush; The outer cap for brush inner lip week to be provided with fore shaft convex, coincide with the fore shaft of the interior cap for brush recessed coupling, to work in closing a cap for brush and fixing in the cap for brush be integrated and open pen container mouth, and play proposition ink brush.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610404342.9A 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 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 |
| CN201610404343.3A CN106092703B (en) | 2014-09-26 | 2014-09-26 | Application of the easy to operate groupization pen in the detection of cell climbing sheet immunohistochemistry |
| 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 |
| CN201410500701.1A CN104359741B (en) | 2014-09-26 | 2014-09-26 | Application of multi-functional PAP pen in cell climbing immunohistochemical detection |
Applications Claiming Priority (1)
| 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 |
Related Child Applications (4)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610404345.2A Division 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 |
| CN201610404344.8A Division 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 |
| CN201610404343.3A Division 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 Division 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 |
Publications (2)
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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 |
| CN106248464A (en) * | 2016-09-02 | 2016-12-21 | 四川大学 | A kind of efficient dyeing apparatus of cell climbing sheet |
| 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 |
| CN109100503A (en) * | 2018-09-20 | 2018-12-28 | 同济大学 | A kind of immunohistochemistry liquid and preparation method thereof |
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| 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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Also Published As
| Publication number | Publication date |
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| CN104359741B (en) | 2017-01-11 |
| CN105938065B (en) | 2019-03-12 |
| CN106092703A (en) | 2016-11-09 |
| CN106092703B (en) | 2018-07-17 |
| CN105938065A (en) | 2016-09-14 |
| CN105938064B (en) | 2019-03-19 |
| CN105938064A (en) | 2016-09-14 |
| CN106092702B (en) | 2018-07-06 |
| CN106092702A (en) | 2016-11-09 |
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