CN106092702A - The application in cell climbing sheet SABC detects of the groupization pen of minimizing antibody consumption - Google Patents
The application in cell climbing sheet SABC detects of the groupization pen of minimizing antibody consumption Download PDFInfo
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- CN106092702A CN106092702A CN201610404342.9A CN201610404342A CN106092702A CN 106092702 A CN106092702 A CN 106092702A CN 201610404342 A CN201610404342 A CN 201610404342A CN 106092702 A CN106092702 A CN 106092702A
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- 230000009194 climbing Effects 0.000 title claims abstract description 26
- 238000004043 dyeing Methods 0.000 claims abstract description 14
- 239000011148 porous material Substances 0.000 claims abstract description 10
- 230000001744 histochemical effect Effects 0.000 claims abstract description 7
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 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
- 230000000694 effects Effects 0.000 claims description 10
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims description 10
- 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
- 238000013115 immunohistochemical detection Methods 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
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 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
- 239000007791 liquid phase Substances 0.000 claims description 3
- 229920002866 paraformaldehyde Polymers 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
- 230000008878 coupling Effects 0.000 claims 1
- 238000010168 coupling process Methods 0.000 claims 1
- 238000005859 coupling reaction Methods 0.000 claims 1
- 239000011435 rock Substances 0.000 claims 1
- 238000011532 immunohistochemical staining Methods 0.000 abstract description 7
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 abstract description 6
- 238000009792 diffusion process Methods 0.000 abstract description 4
- 238000012797 qualification Methods 0.000 abstract description 3
- 239000011521 glass Substances 0.000 abstract description 2
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 6
- 238000000034 method Methods 0.000 description 4
- 238000002372 labelling Methods 0.000 description 3
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 238000003365 immunocytochemistry Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 150000002576 ketones Chemical class 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
- 238000011160 research Methods 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 238000012360 testing method 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
- 238000005096 rolling process Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000000126 substance Substances 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
-
- 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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- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
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- Analytical Chemistry (AREA)
- Urology & Nephrology (AREA)
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Abstract
The invention discloses the application in cell climbing sheet SABC detects of a kind of groupization pen reducing antibody consumption, be inoculated in by cell in microscope slide formula culture dish and carry out cell climbing sheet, cell is cultivated after terminating and is 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.The present invention is applicable to the various immunohistochemical stainings experiment of glass slice, can substantially reduce antibody and reagent dosage, it is to avoid liquid trickling and diffusion during dyeing, improves speed of operation.It is especially applicable in the experimentation of cell climbing sheet or cell smear carrying out on a large scale, the immunohistochemical staining of the many groups of large sample.
Description
The application is application number: 201410500701.1, the applying date: 2014.9.26, title: " multipurpose SABC pen
Application in cell climbing sheet Immunohistochemical detection " divisional application.
Technical field
The present invention relates to the application in cell climbing sheet Immunohistochemical detection of a kind of SABC pen.
Background technology
By specific antibody, labelling and the content thereof of some chemical constituents analysis in tissue slice and cell climbing sheet thereof are entered
Row organizes, location qualitative with cell in-situ or quantitative study, and this technology is referred to as immunocytochemistry
(immunocytochemistry) technology.
Generally cell climbing sheet Immunohistochemical detection is individual cell climbing sheet to carry 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 science of multisample and many groups is real
Test scientific research.
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 can be substantially reduced
And reagent dosage, it is to avoid liquid trickling and diffusion during dyeing, improve the multipurpose SABC pen of speed of operation at cell climbing sheet
Application in Immunohistochemical detection.
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: connect by cell
Planting and carry out cell climbing sheet in microscope slide formula culture dish, cell is cultivated after terminating and is 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 length corresponding with microscope slide size
Square box trace line chase, can deviate from the bottom of ware body along rectangle frame trace line chase 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%, ring
Ketone 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 the most recessed with sharp keen cutter
Ditch is drawn, is hooked, and then adds in ware bottom surface and can be deviate from by the single hole bottom land of ware body by external force.
Described SABC pen includes pen container, arranges spongioid cylinder pen core, be perfused with ink in pen container in pen container,
Spongioid cylinder pen core front end arranges wooden pen core；Spongioid cylinder pen core is placed in pen container, and pen container is rectangular
Body or cylinder, have 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, shakes
Make to mix the effect of ink during rolling pen container；The described interior cap for brush rise closing pen container mouth, prevent ink volatilization keep its liquid phase state and
Couple the effect of the outer cap for brush, its inner surface have and pen container tone close female thread, can spiral cover pen container mouth, the outer surface of the interior cap for brush sets
Have mate with outer cap for brush inner lip coincide fore shaft recessed, cylinder pen core end side is provided with ink brush；The inner lip of the outer cap for brush
It is recessed that week 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 in cultivation
Ware is the bevel-faced form of the angle cooperation on inclined-plane；Culture dish lid and the interleave depth >=10mm of culture dish anastomotic stoma periphery.
The present invention is conducive to the dyeing of SABC, and staining procedure is similar to Immunohistochemistry, but sample process
Temperature, the time, the standard such as reagent concentration is consistent, and an anti-kind of labelling is many, and comparability and reliability are significantly increased！Process
Sample size many, efficient quick.PBS, sera incubation, labelling two resist, and lining dye (haematoxylin) etc. need not the reality separated
Testing operation can synchronize to process, convenient and swift, experiment condition standard is consistent and easy to control.Large area cell climbing sheet is used to carry out
Cell cultivates its temperature processed, time, and the standard such as reagent concentration is consistent, makes the comparability of experimental result and reliability the most aobvious
Write and improve.The quantity of Tissue Culture Dish hole slot reduces, easy to operate, efficient quick.Specific ink formulations has fully ensured that work
Make effect.It is applicable to the experiment of various immunohistochemical staining, including；Paraffin tissue sections, frozen tissue section and cell are climbed
The immunohistochemical staining experiment of sheet, can substantially reduce antibody and reagent dosage, it is to avoid liquid trickling and diffusion during dyeing, carries
High speed of operation；It is especially applicable in the experimentation of cell climbing sheet or cell smear carrying out on a large scale, 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 stainings are tested, and can substantially reduce antibody and reagent dosage, it is to avoid liquid trickling and diffusion during dyeing, carry
High 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 load by cell
Carrying out cell climbing sheet in slide formula culture dish, cell is cultivated after terminating and is carried out immunocyte histochemical stain qualification；Described load
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
More than microscope slide, 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 corresponding with microscope slide size
Frame trace line chase 4, can deviate from the bottom of ware body along rectangle frame trace line chase 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%, ring
Ketone 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 the most recessed with sharp keen cutter
Ditch is drawn, is hooked, and then adds in ware bottom surface and can be deviate from by the single hole bottom land of ware body by external force.
Described SABC pen includes pen container 5, arranges spongioid cylinder pen core 6, be perfused with oil in pen container in pen container
Ink, 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, have 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, make to mix the effect of ink；The described interior cap for brush plays closing pen container mouth, prevents ink volatilization from keeping its liquid phase
State and couple the outer cap for brush effect, its inner surface have and pen container tone conjunction female thread, can spiral cover pen container mouth, outside the interior cap for brush
Surface 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；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, rise in closing the cap for brush and fixing in the cap for brush be integrated out
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 in cultivation
Ware is the bevel-faced form of the angle cooperation on inclined-plane；Culture dish lid and the interleave depth >=10mm of culture dish anastomotic stoma periphery.
Claims (4)
1. reduce the application in cell climbing sheet SABC detects of the groupization pen of antibody consumption, it is characterized in that: by cell
Being inoculated in microscope slide formula culture dish and carry out cell climbing sheet, cell is cultivated after terminating and is carried out immunocyte histochemical stain mirror
Fixed；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 hole
Room 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 corresponding with microscope slide size
Rectangle frame trace line chase, can deviate from the bottom of ware body along rectangle frame trace line chase, it is thus achieved that have cultivation under external force
Cell face, just as at the bottom of the ware of microscope slide, is 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%；
Described SABC pen includes pen container, arranges spongioid cylinder pen core, be perfused with ink, sponge in pen container in pen container
The cylinder pen core front end of sample arranges wooden pen core；Spongioid cylinder pen core is placed in pen container, pen container be 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, rocks pen
Make to mix the effect of ink during cylinder；The described interior cap for brush plays closing pen container mouth, prevents ink volatilization from keeping its liquid phase state and connection
The effect of the outer cap for brush, its inner surface have and pen container tone close female thread, can spiral cover pen container mouth, the outer surface of the interior cap for brush be provided with
The fore shaft that outer cap for brush inner lip coupling is coincide is recessed, and cylinder pen core end side is provided with ink brush；The inner lip week of the outer cap for brush sets
Having fore shaft convex, mate recessed with the fore shaft of the interior cap for brush, 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；
Before the bottom of ware body can being deviate from along rectangle frame trace line chase under external force, first with sharp keen cutter chase along the line
Draw, hook, then the single hole bottom land of ware body can be deviate from by external force a little in ware bottom surface.
The application in cell climbing sheet SABC detects of the groupization pen of minimizing antibody consumption the most according to claim 1,
It is characterized in that: an angle of many pore chambers culture dish is bevel-faced form, facilitates bearing mark.
The application in cell climbing sheet SABC detects of the groupization pen of minimizing antibody consumption the most according to claim 2,
It is characterized in that: an angle of culture dish lid is in the bevel-faced form coordinated with the angle that culture dish is inclined-plane.
The application in cell climbing sheet SABC detects of the groupization pen of minimizing antibody consumption the most according to claim 3,
It is characterized in that: culture dish lid and the interleave depth >=10mm of culture dish anastomotic stoma periphery.
Priority Applications (1)
| 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 |
Applications Claiming Priority (2)
| 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 |
| CN201410500701.1A CN104359741B (en) | 2014-09-26 | 2014-09-26 | Application of multi-functional PAP pen in cell climbing immunohistochemical detection |
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 |
|---|---|
| CN106092702A true CN106092702A (en) | 2016-11-09 |
| CN106092702B CN106092702B (en) | 2018-07-06 |
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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 |
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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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| CN106092703B (en) | 2018-07-17 |
| CN104359741A (en) | 2015-02-18 |
| CN105938064B (en) | 2019-03-19 |
| CN106092702B (en) | 2018-07-06 |
| CN106092703A (en) | 2016-11-09 |
| CN105938065B (en) | 2019-03-12 |
| CN105938064A (en) | 2016-09-14 |
| CN104359741B (en) | 2017-01-11 |
| CN105938065A (en) | 2016-09-14 |
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