CN107860632A - A kind of drosophila wing bud active immunity group makeup is put - Google Patents
A kind of drosophila wing bud active immunity group makeup is put Download PDFInfo
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- CN107860632A CN107860632A CN201711087015.6A CN201711087015A CN107860632A CN 107860632 A CN107860632 A CN 107860632A CN 201711087015 A CN201711087015 A CN 201711087015A CN 107860632 A CN107860632 A CN 107860632A
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- 241000255581 Drosophila <fruit fly, genus> Species 0.000 title claims abstract description 13
- 230000036039 immunity Effects 0.000 title claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 84
- 239000012530 fluid Substances 0.000 claims abstract description 83
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 60
- 239000006059 cover glass Substances 0.000 claims abstract description 12
- 239000002699 waste material Substances 0.000 claims abstract description 11
- 238000011534 incubation Methods 0.000 claims description 19
- 239000000523 sample Substances 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 abstract description 7
- 238000004043 dyeing Methods 0.000 abstract description 5
- 230000002055 immunohistochemical effect Effects 0.000 abstract description 3
- 238000011532 immunohistochemical staining Methods 0.000 abstract description 2
- 238000005406 washing Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 6
- 239000007853 buffer solution Substances 0.000 description 3
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 229940099259 vaseline Drugs 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
- G01N1/312—Apparatus therefor for samples mounted on planar substrates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
A kind of drosophila wing bud active immunity group makeup is put, and is related to biological tissue's antibody or fluorescent dyeing equipment.The present invention solves existing drosophila wing bud immunohistochemical experiment manual operation, the technical problem of time and effort consuming.The present invention includes slide, cover glass, primary antibody fluid reservoir, secondary antibody fluid reservoir, PBT cushioning liquid fluid reservoir, formaldehyde fixer fluid reservoir, feed tube, drain pipe, some connecting tubes and waste liquid tank；The slide is provided with rectangle culture chamber, and the cover glass is located above rectangle culture chamber, and the rectangle culture chamber side side wall is provided with some culture chamber inlets, and the rectangle culture chamber opposite side side wall is provided with some culture chamber liquid outlets.The present invention uses special slide, is tested suitable for the immunohistochemical staining of relatively thin biological tissue, can be automatically performed SABC flow, is accurately controlled the time of each step dyeing and washing, saves time and efforts, ensure that the quality of experiment.
Description
Technical field
The invention belongs to biological tissue's culture antibody or fluorescent dyeing equipment technical field, more particularly to a kind of drosophila
The makeup of wing bud active immunity group is put.
Background technology
SABC, is applied immunology general principle --- antigen-antibody reaction, i.e. antigen is combined with antibody specificity
Principle, the developer fluorescein of labelled antibody, enzyme, metal ion, isotope colour developing is determined tissue by chemical reaction
Intracellular antigen peptide and protein, it is positioned, be qualitative and determine quantifier elimination.
The immunohistochemical experiment of drosophila wing bud includes many steps, and key step has：Drosophila larvae is dissected, wing bud tissue
40~60 minutes are fixed in formaldehyde fixer, then washes 1 hour of fixer；Tissue is placed in certain density primary antibody again
In, it is incubated 8~12 hours under low temperature (4~6 DEG C), is washed out primary antibody；Add two anti-dye 1 hour, wash secondary antibody.Entirely
Flow not only complex steps, and per, it is necessary to be manually loaded and change clothes liquid, expending time and efforts, such as at regular intervals
The some steps of fruit miss loading time or dyeing time is long, can all influence final SABC effect.
The content of the invention
The shortcomings that it is an object of the invention to overcome prior art, there is provided a kind of drosophila wing bud active immunity group makeup is put,
Solve existing drosophila wing bud immunohistochemical experiment manual operation, the technical problem of time and effort consuming.
The present invention is achieved by the following technical solutions：
A kind of drosophila wing bud active immunity group makeup is put, wherein：Including slide, cover glass, primary antibody fluid reservoir, secondary antibody storage
Flow container, PBT cushioning liquid fluid reservoir, formaldehyde fixer fluid reservoir, feed tube, drain pipe, some connecting tubes and waste liquid tank；
The slide is provided with rectangle and is incubated chamber, and the cover glass is located at rectangle and is incubated above chamber, and the rectangle is incubated chamber
Side side wall is provided with some incubation chamber inlets, and the rectangle is incubated chamber opposite side side wall and goes out liquid provided with some incubation chambers
Mouthful；
The primary antibody fluid reservoir, secondary antibody fluid reservoir, PBT cushioning liquid fluid reservoir and formaldehyde fixer fluid reservoir are located at load glass
Above piece side, the waste liquid tank is located at below slide opposite side, and the primary antibody fluid reservoir, secondary antibody fluid reservoir, PBT bufferings are molten
The outlet of liquid fluid reservoir, formaldehyde fixer fluid reservoir is connected by connecting tube with feed tube respectively, and the feed tube passes through some
Chamber inlet is incubated to be connected with rectangle incubation chamber；Some incubation chamber liquid outlets are connected by drain pipe with waste liquid tank；
The primary antibody fluid reservoir, secondary antibody fluid reservoir, PBT cushioning liquid fluid reservoir, formaldehyde fixer fluid reservoir and feed tube
Between connecting tube on be equipped with by single chip control module control minisize liquid control valve, the drain pipe be provided with by list
The drawing liquid pump of piece machine control module control；
The primary antibody fluid reservoir, secondary antibody fluid reservoir and incubation chamber are provided with the temperature control controlled by single chip control module
Device processed, the slide are provided with the infrared temperature probe that detection rectangle is incubated chamber temperature.
Further, the incubation chamber inlet has 2~5；The quantity for being incubated chamber liquid outlet and being incubated chamber inlet
Unanimously.
Cover glass in the present invention can realize that rectangle is incubated the sealing of chamber, avoid external microbe from infecting Incubating Solution；It is described
Primary antibody fluid reservoir, secondary antibody fluid reservoir, PBT cushioning liquid fluid reservoir, formaldehyde fixer fluid reservoir, connecting tube, minisize liquid control
The setting of valve, feed tube, drain pipe and waste liquid tank, the Incubating Solution that can be incubated intracavitary to rectangle as needed be changed, especially
It is that minisize liquid control valve is provided between formaldehyde fixer, primary antibody, secondary antibody, PBT cushioning liquid, fluid reservoir and feed tube,
And drawing liquid pump has been put on drain pipe, it can be achieved to be automatically brought into operation；The setting of low-temperature control device on primary antibody, secondary antibody fluid reservoir, carry
Slide is provided with infrared temperature probe, it is ensured that is incubated at low temperature during wing bud one anti-dye of tissue.
The present invention uses special slide, is tested suitable for the immunohistochemical staining of relatively thin biological tissue, can be automatic
SABC flow is completed, the time of each step dyeing and washing is accurately controlled, saves time and efforts, ensure that experiment
Quality.
Brief description of the drawings
Fig. 1 is the structural representation that a kind of drosophila wing bud active immunity group makeup is put；
Fig. 2 is the structural representation of slide, cover glass；
Fig. 3 is Fig. 2 top view；
Fig. 4 is single chip machine controlling circuit theory diagram.
Embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples.
As shown in Figure 1 to Figure 3, a kind of drosophila wing bud active immunity group makeup in the present embodiment is put, including slide 1,
Cover glass 2, primary antibody fluid reservoir 10, secondary antibody fluid reservoir 11, PBT cushioning liquid fluid reservoir 12, formaldehyde fixer fluid reservoir 13, feed liquor
Pipe 4, drain pipe 5, some connecting tubes 3 and waste liquid tank 6；
The slide 1 is provided with rectangle and is incubated chamber 7, and the cover glass 2 is located at rectangle and is incubated the top of chamber 7, and the rectangle is incubated
Educate the side side wall of chamber 7 and be provided with some incubation chamber inlets 8, the rectangle is incubated the opposite side side wall of chamber 7 and is provided with some incubations
Chamber liquid outlet 9；
The primary antibody fluid reservoir 10, secondary antibody fluid reservoir 11, PBT cushioning liquid fluid reservoir 12 and formaldehyde fixer fluid reservoir 13
It is located above the side of slide 1, the waste liquid tank 6 is located at below the opposite side of slide 1, the primary antibody fluid reservoir 10, secondary antibody storage
The outlet of flow container 11, PBT cushioning liquid fluid reservoir 12, formaldehyde fixer fluid reservoir 13 is connected by connecting tube 3 and feed tube 4 respectively
Connect, the feed tube 4 is incubated chamber 7 with rectangle by some incubation chamber inlets 8 and is connected；Some incubation chamber liquid outlets 9 are logical
Drain pipe 5 is crossed to be connected with waste liquid tank 6；
The primary antibody fluid reservoir 10, secondary antibody fluid reservoir 11, PBT cushioning liquid fluid reservoir 12, formaldehyde fixer fluid reservoir 13
Be equipped with connecting tube 3 between feed tube 4 by single chip control module control minisize liquid control valve 14, it is described go out liquid
Pipe 5 is provided with the drawing liquid pump 15 controlled by single chip control module.
The primary antibody fluid reservoir, secondary antibody fluid reservoir are provided with the low-temperature control device controlled by single chip control module, institute
State slide 1 and be provided with infrared temperature probe 16, detection rectangle is incubated the temperature of chamber 7.
The incubation chamber inlet has 2~5 (the present embodiment is 2)；The incubation chamber liquid outlet is with being incubated chamber feed liquor
The quantity of mouth is consistent.
As shown in figure 4, the primary antibody fluid reservoir 10, secondary antibody fluid reservoir 11, PBT cushioning liquid fluid reservoir 12, formaldehyde are fixed
The minisize liquid control valve 14 and drawing liquid pump 15 set in connecting tube 3 between liquid fluid reservoir 13 and feed tube 4, and single-chip microcomputer
Control module connects, so as to realize operation automation.
As shown in figure 4, the infrared temperature probe 16 is connected with single chip control module, slide is measured in real time and is incubated chamber
Interior temperature, the primary antibody fluid reservoir 10, the low-temperature control device of secondary antibody fluid reservoir 11 are connected with single chip control module, infrared
Temperature probe surveys data feedback to single-chip microcomputer, and single-chip microcomputer controls the low temperature control of primary antibody fluid reservoir 10, secondary antibody fluid reservoir 11 again
Device, so that it is guaranteed that when primary antibody solution is incubated, the temperature for being incubated chamber is 4~6 DEG C, other staining procedures, is incubated the temperature of chamber
Control in room temperature.
The use process of the present invention：
It is incubated in use, the wing bud that gets off will be dissected in phosphate buffer and be transferred to rectangle in chamber 7, covered 2,
It is and with resinene or vaseline that cover glass 2 is sealed around；
Pass through single chip machine controlling circuit：
The minisize liquid control valve 14 and drawing liquid pump 15 being first turned in the lower section connecting tube of formaldehyde fixer fluid reservoir 13, really
Slide reservoir is protected full of the minisize liquid control valve after fixer, closed in the lower section connecting tube of formaldehyde fixer fluid reservoir 13
14 and drawing liquid pump 15, fix 40~60min；
Then, the minisize liquid control valve 14 and drawing liquid pump 15 in the lower section connecting tube of PBT cushioning liquid fluid reservoir 12 are opened,
After buffer solution persistently cleans 1~1.5h of formaldehyde fixer, the miniature liquid in the lower section connecting tube of closing PBT cushioning liquid fluid reservoir 12
Body control valve 14 and drawing liquid pump 15；
The minisize liquid control valve 14 and drawing liquid pump 15 in the lower section connecting tube of primary antibody fluid reservoir 10 are opened simultaneously, it is ensured that primary antibody
Cryogenic fluid is closed micro- in the lower section connecting tube of primary antibody fluid reservoir 10 full of slide reservoir and fully after immersion wing bud tissue
Type liquid control valve 14 and drawing liquid pump 15, low temperature are incubated 8~12h.
The minisize liquid control valve 14 and drawing liquid pump 15 being again turned in the lower section connecting tube of PBT cushioning liquid fluid reservoir 12,
After buffer solution persistently cleans 1~1.5h of primary antibody solution, the minisize liquid in the lower section connecting tube of closing PBT cushioning liquid fluid reservoir 12
Control valve 14 and drawing liquid pump 15；
The drawing liquid pump in the minisize liquid control valve 14 and drain pipe 5 in the lower section connecting tube of secondary antibody fluid reservoir 11 is opened simultaneously
15, it is ensured that secondary antibody cryogenic fluid closes the lower section of secondary antibody fluid reservoir 10 full of slide reservoir and fully after immersion wing bud tissue
Minisize liquid control valve 14 and drawing liquid pump 15 in connecting tube, low temperature are incubated 1~1.2h.
Finally open in the minisize liquid control valve 14 and drain pipe 5 in the lower section connecting tube of PBT cushioning liquid fluid reservoir 12
Drawing liquid pump 15, after buffer solution persistently cleans 1~1.5h of two corresponding anti-solution, close the lower section connecting tube of PBT cushioning liquid fluid reservoir 12
On minisize liquid control valve 14 and drawing liquid pump 15, complete experiment.
After the completion of experiment, slide is directly placed and taken pictures under fluorescence microscope or laser confocal microscope.
The cover glass is covering incubation chamber, and with after the adhesive seal surrounding such as resinene, is incubated chamber, is incubated chamber
Inlet, is incubated chamber liquid outlet and connected feed tube, drain pipe form closed space, so that drawing liquid pump will can be incubated
The liquid of intracavitary is taken away.
The present invention can be embodied without departing from the spiritual or substantive of invention in a variety of forms, it should therefore be appreciated that on
State embodiment and be not limited to foregoing details, and should widely be explained in claim limited range, therefore fall into right
It is required that or the change in its equivalent scope and remodeling all should be claim and covered.
Claims (2)
1. a kind of drosophila wing bud active immunity group makeup is put, it is characterised in that：Including slide (1), cover glass (2), primary antibody storage
Flow container (10), secondary antibody fluid reservoir (11), PBT cushioning liquid fluid reservoir (12), formaldehyde fixer fluid reservoir (13), feed tube (4),
Drain pipe (5), some connecting tubes (3) and waste liquid tank (6)；
The slide (1) is provided with rectangle and is incubated chamber (7), and the cover glass (2) is located at rectangle and is incubated above chamber (7), the square
Shape is incubated chamber (7) side side wall and is provided with some incubation chamber inlets (8), and the rectangle is incubated in chamber (7) opposite side side wall and set
There are some incubation chamber liquid outlets (9)；
The primary antibody fluid reservoir (10), secondary antibody fluid reservoir (11), PBT cushioning liquid fluid reservoir (12) and formaldehyde fixer fluid reservoir
(13) it is located above slide (1) side, the waste liquid tank (6) is located at below slide (1) opposite side, the primary antibody fluid reservoir
(10), the outlet of secondary antibody fluid reservoir (11), PBT cushioning liquid fluid reservoir (12), formaldehyde fixer fluid reservoir (13) passes through respectively
Connecting tube (3) is connected with feed tube (4), and the feed tube (4) is incubated chamber (7) by some incubation chamber inlets (8) with rectangle
Connection；Some incubation chamber liquid outlets (9) are connected by drain pipe (5) with waste liquid tank (6)；
The primary antibody fluid reservoir (10), secondary antibody fluid reservoir (11), PBT cushioning liquid fluid reservoir (12), formaldehyde fixer fluid reservoir
(13) the minisize liquid control valve controlled by single chip control module is equipped with the connecting tube (3) between feed tube (4)
(14), the drain pipe (5) is provided with the drawing liquid pump (15) controlled by single chip control module；
The primary antibody fluid reservoir (10), secondary antibody fluid reservoir (12) are provided with controls dress by the low temperature of single chip control module control
Put, the slide (1) is provided with the infrared temperature probe (16) that detection rectangle is incubated chamber (7) temperature.
A kind of 2. biological tissue's culture apparatus according to claim 1, it is characterised in that：The incubation chamber inlet (8)
There are 2~5；The incubation chamber liquid outlet (9) is consistent with the quantity for being incubated chamber inlet (8).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711087015.6A CN107860632B (en) | 2017-11-07 | 2017-11-07 | Automatic immunohistochemical device of fruit bat wing bud |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711087015.6A CN107860632B (en) | 2017-11-07 | 2017-11-07 | Automatic immunohistochemical device of fruit bat wing bud |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107860632A true CN107860632A (en) | 2018-03-30 |
| CN107860632B CN107860632B (en) | 2020-01-03 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711087015.6A Active CN107860632B (en) | 2017-11-07 | 2017-11-07 | Automatic immunohistochemical device of fruit bat wing bud |
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| Country | Link |
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| CN (1) | CN107860632B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109254160A (en) * | 2018-11-09 | 2019-01-22 | 上海交通大学医学院附属第九人民医院 | A kind of automatic Western Blot antibody incubation box |
| CN111006041A (en) * | 2018-10-05 | 2020-04-14 | 伊鲁米那股份有限公司 | Multi-valve fluid cartridge |
| US12000854B2 (en) | 2022-06-02 | 2024-06-04 | Illumina, Inc. | Multi-valve fluid cartridge |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111006041A (en) * | 2018-10-05 | 2020-04-14 | 伊鲁米那股份有限公司 | Multi-valve fluid cartridge |
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| US12000854B2 (en) | 2022-06-02 | 2024-06-04 | Illumina, Inc. | Multi-valve fluid cartridge |
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| Publication number | Publication date |
|---|---|
| CN107860632B (en) | 2020-01-03 |
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