CN107860632B - Automatic immunohistochemical device of fruit bat wing bud - Google Patents
Automatic immunohistochemical device of fruit bat wing bud Download PDFInfo
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- CN107860632B CN107860632B CN201711087015.6A CN201711087015A CN107860632B CN 107860632 B CN107860632 B CN 107860632B CN 201711087015 A CN201711087015 A CN 201711087015A CN 107860632 B CN107860632 B CN 107860632B
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- 230000002055 immunohistochemical effect Effects 0.000 title claims abstract description 16
- 240000003380 Passiflora rubra Species 0.000 title claims description 3
- 210000000007 bat wing Anatomy 0.000 title claims description 3
- 235000013399 edible fruits Nutrition 0.000 title claims description 3
- 239000007788 liquid Substances 0.000 claims abstract description 178
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 60
- 239000000243 solution Substances 0.000 claims abstract description 35
- 239000011521 glass Substances 0.000 claims abstract description 24
- 239000007977 PBT buffer Substances 0.000 claims abstract description 22
- 239000006059 cover glass Substances 0.000 claims abstract description 12
- 239000002699 waste material Substances 0.000 claims abstract description 11
- 241000255588 Tephritidae Species 0.000 claims abstract description 9
- 238000011534 incubation Methods 0.000 claims description 51
- 239000000523 sample Substances 0.000 claims description 8
- 238000002474 experimental method Methods 0.000 abstract description 9
- 238000005406 washing Methods 0.000 abstract description 5
- 238000000034 method Methods 0.000 abstract description 4
- 238000004043 dyeing Methods 0.000 abstract description 3
- 238000010186 staining Methods 0.000 abstract description 3
- 239000007850 fluorescent dye Substances 0.000 abstract description 2
- 238000011532 immunohistochemical staining Methods 0.000 abstract description 2
- 230000012447 hatching Effects 0.000 description 5
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 2
- 238000009529 body temperature measurement Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 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
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram 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
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229940099259 vaseline Drugs 0.000 description 1
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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)
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Abstract
An automatic immunohistochemical device for fruit fly wing buds relates to biological tissue antibody or fluorescent dye dyeing equipment. The invention solves the technical problems of time and labor consumption of manual operation of the conventional fruit fly wing bud immunohistochemical experiment. The invention comprises a glass slide, a cover glass, a primary antibody liquid storage tank, a secondary antibody liquid storage tank, a PBT buffer solution liquid storage tank, a formaldehyde fixing solution liquid storage tank, a liquid inlet pipe, a liquid outlet pipe, a plurality of connecting pipes and a waste liquid tank; the slide glass is equipped with the rectangle and cultivates the chamber, the cover glass is established in rectangle cultivation chamber top, rectangle cultivation chamber one side lateral wall is equipped with a plurality of cultivation chamber inlets, rectangle cultivation chamber opposite side lateral wall is equipped with a plurality of cultivation chamber liquid outlets. The invention adopts the specially-made glass slide, is suitable for the immunohistochemical staining experiment of thinner biological tissues, can automatically complete the immunohistochemical process, accurately control the staining and washing time of each step, save time and energy and ensure the quality of the experiment.
Description
Technical Field
The invention belongs to the technical field of biological tissue culture antibody or fluorescent dye dyeing equipment, and particularly relates to an automatic immunohistochemical device for drosophila wing buds.
Background
Immunohistochemistry is the research of determining the antigen polypeptide and protein in tissue cell by applying the basic principle of immunology, namely the antigen-antibody reaction, namely the principle of specific combination of antigen and antibody, and developing the color developing agent fluorescein, enzyme, metal ion and isotope of the labeled antibody through chemical reaction, and carrying out positioning, qualitative and quantitative research on the antigen polypeptide and protein.
The immunohistochemical experiment of the fruit fly wing bud comprises a plurality of steps, and the main steps comprise: dissecting drosophila larvae, fixing wing bud tissues in formaldehyde fixing solution for 40-60 minutes, and then washing the fixing solution for 1 hour; placing the tissue in primary antibody with a certain concentration, incubating for 8-12 hours at a low temperature (4-6 ℃), and then washing the primary antibody; secondary antibody was added for staining for 1 hour and the secondary antibody was washed. The whole process is complex in steps, manual sample adding and liquid changing and washing are needed at regular intervals, time and energy are consumed, and if some steps miss sample adding time or dyeing time is too long, the final immunohistochemical effect can be influenced.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides an automatic immunohistochemical device for fruit fly wing buds, and solves the technical problems of time and labor consumption caused by manual operation of the conventional fruit fly wing bud immunohistochemical experiment.
The invention is realized by the following technical scheme:
an automatic immunohistochemical device of fruit fly wing bud, wherein: comprises a glass slide, a cover glass, a primary antibody liquid storage tank, a secondary antibody liquid storage tank, a PBT buffer solution liquid storage tank, a formaldehyde fixing liquid storage tank, a liquid inlet pipe, a liquid outlet pipe, a plurality of connecting pipes and a waste liquid tank;
the glass slide is provided with a rectangular incubation cavity, the cover glass is arranged above the rectangular incubation cavity, a plurality of incubation cavity liquid inlets are formed in the side wall of one side of the rectangular incubation cavity, and a plurality of incubation cavity liquid outlets are formed in the side wall of the other side of the rectangular incubation cavity;
the first anti-liquid storage tank, the second anti-liquid storage tank, the PBT buffer solution storage tank and the formaldehyde fixing solution storage tank are arranged above one side of the glass slide, the waste liquid tank is arranged below the other side of the glass slide, outlets of the first anti-liquid storage tank, the second anti-liquid storage tank, the PBT buffer solution storage tank and the formaldehyde fixing solution storage tank are respectively connected with a liquid inlet pipe through connecting pipes, and the liquid inlet pipe is connected with the rectangular incubation cavity through liquid inlets of the incubation cavities; liquid outlets of the incubation cavities are connected with a waste liquid tank through liquid outlet pipes;
the connecting pipes among the primary anti-liquid storage tank, the secondary anti-liquid storage tank, the PBT buffer solution storage tank, the formaldehyde stationary liquid storage tank and the liquid inlet pipe are all provided with miniature liquid control valves controlled by the singlechip control module, and the liquid outlet pipe is provided with a liquid pump controlled by the singlechip control module;
the first anti-liquid storage tank, the second anti-liquid storage tank and the incubation cavity are provided with temperature control devices controlled by a single chip microcomputer control module, and the glass slide is provided with an infrared temperature measuring probe for detecting the temperature of the rectangular incubation cavity.
Furthermore, 2-5 liquid inlets of the incubation cavity are provided; the liquid outlets of the incubation cavity and the liquid inlet of the incubation cavity are consistent in quantity.
The cover glass can realize the sealing of the rectangular incubation cavity, so that the incubation liquid is prevented from being infected by external microorganisms; the arrangement of the primary anti-liquid storage tank, the secondary anti-liquid storage tank, the PBT buffer solution storage tank, the formaldehyde stationary liquid storage tank, the connecting pipe, the micro liquid control valve, the liquid inlet pipe, the liquid outlet pipe and the waste liquid tank can replace the incubation liquid in the rectangular incubation cavity as required, particularly, the micro liquid control valve is arranged between the formaldehyde stationary liquid, the primary anti-liquid, the secondary anti-liquid, the PBT buffer solution storage tank and the liquid inlet pipe, and the liquid pump is arranged on the liquid outlet pipe, so that the automatic operation can be realized; the arrangement of the low-temperature control device on the primary antibody and secondary antibody liquid storage tanks and the arrangement of the infrared temperature measuring probe on the glass slide ensure that the fin bud tissue is incubated at low temperature when the primary antibody is dyed.
The invention adopts the specially-made glass slide, is suitable for the immunohistochemical staining experiment of thinner biological tissues, can automatically complete the immunohistochemical process, accurately control the staining and washing time of each step, save time and energy and ensure the quality of the experiment.
Drawings
FIG. 1 is a schematic structural view of an automatic immunohistochemical apparatus for fruit fly wing buds;
FIG. 2 is a schematic view of a slide and cover slip configuration;
FIG. 3 is a top view of FIG. 2;
fig. 4 is a schematic block diagram of a single chip microcomputer control circuit.
Detailed Description
The invention is described in further detail below with reference to the figures and examples.
As shown in fig. 1 to 3, the automatic immunohistochemical device for fruit fly wing buds in the present embodiment includes a glass slide 1, a cover glass 2, a primary antibody liquid storage tank 10, a secondary antibody liquid storage tank 11, a PBT buffer solution liquid storage tank 12, a formaldehyde fixing liquid storage tank 13, a liquid inlet pipe 4, a liquid outlet pipe 5, a plurality of connecting pipes 3, and a waste liquid tank 6;
the glass slide 1 is provided with a rectangular incubation cavity 7, the cover glass 2 is arranged above the rectangular incubation cavity 7, a plurality of incubation cavity liquid inlets 8 are formed in the side wall of one side of the rectangular incubation cavity 7, and a plurality of incubation cavity liquid outlets 9 are formed in the side wall of the other side of the rectangular incubation cavity 7;
the first anti-liquid storage tank 10, the second antibody liquid storage tank 11, the PBT buffer solution liquid storage tank 12 and the formaldehyde fixing liquid storage tank 13 are arranged above one side of the glass slide 1, the waste liquid tank 6 is arranged below the other side of the glass slide 1, outlets of the first anti-liquid storage tank 10, the second antibody liquid storage tank 11, the PBT buffer solution liquid storage tank 12 and the formaldehyde fixing liquid storage tank 13 are respectively connected with the liquid inlet pipe 4 through the connecting pipe 3, and the liquid inlet pipe 4 is connected with the rectangular incubation cavity 7 through a plurality of incubation cavity liquid inlets 8; the liquid outlets 9 of the incubation cavities are connected with a waste liquid tank 6 through a liquid outlet pipe 5;
and the connecting pipes 3 between the first anti-liquid storage tank 10, the second anti-liquid storage tank 11, the PBT buffer solution storage tank 12, the formaldehyde fixing solution storage tank 13 and the liquid inlet pipe 4 are respectively provided with a micro liquid control valve 14 controlled by a singlechip control module, and the liquid outlet pipe 5 is provided with a liquid pump 15 controlled by the singlechip control module.
The first anti-liquid storage tank and the second anti-liquid storage tank are provided with low-temperature control devices controlled by a single-chip microcomputer control module, and the glass slide 1 is provided with an infrared temperature measuring probe 16 for detecting the temperature of the rectangular incubation cavity 7.
2-5 (2 in the embodiment) liquid inlets of the incubation cavity; the liquid outlets of the incubation cavity and the liquid inlet of the incubation cavity are consistent in quantity.
As shown in fig. 4, the micro liquid control valve 14 and the liquid pump 15 arranged on the connecting pipe 3 between the first anti-liquid storage tank 10, the second anti-liquid storage tank 11, the PBT buffer solution storage tank 12, the formaldehyde fixing liquid storage tank 13 and the liquid inlet pipe 4 are all connected with the single chip microcomputer control module, thereby realizing the automation of the operation.
As shown in fig. 4, the infrared temperature measurement probe 16 is connected with the single chip microcomputer control module to measure the temperature in the glass slide incubation cavity in real time, the low temperature control devices of the first-resistance liquid storage tank 10 and the second-resistance liquid storage tank 11 are connected with the single chip microcomputer control module, data measured by the infrared temperature measurement probe are fed back to the single chip microcomputer, and the single chip microcomputer controls the low temperature control devices of the first-resistance liquid storage tank 10 and the second-resistance liquid storage tank 11, so that the temperature of the incubation cavity is ensured to be 4-6 ℃ during incubation of the first-resistance solution, and the temperature of the incubation cavity is controlled at room temperature.
The use process of the invention is as follows:
when in use, the wing buds dissected in the phosphate buffer solution are transferred into the rectangular incubation cavity 7, the cover glass 2 is covered, and the periphery of the cover glass 2 is sealed by neutral resin or vaseline;
through singlechip control circuit:
firstly, opening a micro liquid control valve 14 and a liquid drawing pump 15 on a connecting pipe below a formaldehyde fixing liquid storage tank 13, after ensuring that a glass slide storage tank is filled with fixing liquid, closing the micro liquid control valve 14 and the liquid drawing pump 15 on the connecting pipe below the formaldehyde fixing liquid storage tank 13, and fixing for 40-60 min;
then, the micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the PBT buffer solution storage tank 12 are opened, and after the buffer solution continuously cleans the formaldehyde stationary liquid for 1-1.5 h, the micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the PBT buffer solution storage tank 12 are closed;
and simultaneously, the micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the primary anti-low temperature liquid storage tank 10 are opened to ensure that the primary anti-low temperature solution is fully filled in the slide liquid storage tank and fully soaks the fin bud tissue, the micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the primary anti-low temperature liquid storage tank 10 are closed, and the incubation is carried out for 8-12 hours at low temperature.
The micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the PBT buffer solution storage tank 12 are started again, and after the buffer solution continuously cleans the primary antibody solution for 1-1.5 hours, the micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the PBT buffer solution storage tank 12 are closed;
and simultaneously, the micro liquid control valve 14 on the connecting pipe below the secondary antibody liquid storage tank 11 and the liquid pump 15 on the liquid outlet pipe 5 are opened to ensure that the secondary antibody low-temperature solution is filled in the glass slide liquid storage tank and fully soaks the fin bud tissue, the micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the secondary antibody liquid storage tank 10 are closed, and the incubation is carried out for 1-1.2 hours at low temperature.
And finally, starting the micro liquid control valve 14 on the connecting pipe below the PBT buffer solution storage tank 12 and the liquid pump 15 on the liquid outlet pipe 5, continuously cleaning the secondary antibody solution by the buffer solution for 1-1.5 h, and then closing the micro liquid control valve 14 and the liquid pump 15 on the connecting pipe below the PBT buffer solution storage tank 12 to complete the experiment.
After the experiment is finished, the glass slide is directly placed under a fluorescence microscope or a laser confocal microscope for photographing.
The cover glass is covering and is hatching the chamber to and after using adhesive such as neutral resin to seal all around, hatching the chamber, hatching chamber inlet, hatching chamber liquid outlet and the feed liquor pipe that links to each other, the drain pipe forms inclosed space to the liquid pump can take away the liquid of hatching the intracavity.
As the present invention may be embodied in several forms without departing from the spirit or essential characteristics thereof, it should also be understood that the above-described embodiments are not limited by any of the details of the foregoing description, but rather should be construed broadly within its scope as defined in the appended claims, and therefore all changes and modifications that fall within the meets and bounds of the claims, or equivalences of such meets and bounds are therefore intended to be embraced by the appended claims.
Claims (2)
1. The utility model provides an automatic immunohistochemical device of fruit bat wing bud which characterized in that: comprises a glass slide (1), a cover glass (2), a primary antibody liquid storage tank (10), a secondary antibody liquid storage tank (11), a PBT buffer solution liquid storage tank (12), a formaldehyde stationary liquid storage tank (13), a liquid inlet pipe (4), a liquid outlet pipe (5), a plurality of connecting pipes (3) and a waste liquid tank (6);
the glass slide (1) is provided with a rectangular incubation cavity (7), the cover glass (2) is arranged above the rectangular incubation cavity (7), a plurality of incubation cavity liquid inlets (8) are formed in the side wall of one side of the rectangular incubation cavity (7), and a plurality of incubation cavity liquid outlets (9) are formed in the side wall of the other side of the rectangular incubation cavity (7);
the first anti-liquid storage tank (10), the second antibody liquid storage tank (11), the PBT buffer solution liquid storage tank (12) and the formaldehyde fixing solution liquid storage tank (13) are arranged above one side of the glass slide (1), the waste liquid tank (6) is arranged below the other side of the glass slide (1), outlets of the first anti-liquid storage tank (10), the second antibody liquid storage tank (11), the PBT buffer solution liquid storage tank (12) and the formaldehyde fixing solution liquid storage tank (13) are respectively connected with the liquid inlet pipe (4) through connecting pipes (3), and the liquid inlet pipe (4) is connected with the rectangular incubation cavity (7) through a plurality of incubation cavity liquid inlets (8); the liquid outlets (9) of the incubation cavities are connected with a waste liquid tank (6) through a liquid outlet pipe (5);
a micro liquid control valve (14) controlled by a singlechip control module is arranged on each connecting pipe (3) between the first anti-liquid storage tank (10), the second anti-liquid storage tank (11), the PBT buffer solution storage tank (12), the formaldehyde fixing liquid storage tank (13) and the liquid inlet pipe (4), and a liquid pump (15) controlled by the singlechip control module is arranged on the liquid outlet pipe (5);
the first anti-liquid storage tank (10) and the second anti-liquid storage tank (12) are provided with low-temperature control devices controlled by the singlechip control module， An infrared temperature measuring probe (16) for detecting the temperature of the rectangular incubation cavity (7) is arranged on the glass slide (1).
2. The automatic immunohistochemical device of fruit fly wing bud according to claim 1, characterized in that: 2-5 liquid inlets (8) of the incubation cavity are provided; the number of the incubation cavity liquid outlets (9) is consistent with that of the incubation cavity liquid inlets (8).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201711087015.6A CN107860632B (en) | 2017-11-07 | 2017-11-07 | Automatic immunohistochemical device of fruit bat wing bud |
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| 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 |
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| CN107860632A CN107860632A (en) | 2018-03-30 |
| CN107860632B true CN107860632B (en) | 2020-01-03 |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| NL2021969B1 (en) * | 2018-10-05 | 2020-05-12 | Illumina Inc | Multi-valve fluid cartridge |
| 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 |
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2017
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