CN101126688A - Large scale cell micro specimen preparation method - Google Patents
Large scale cell micro specimen preparation method Download PDFInfo
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- CN101126688A CN101126688A CNA2007100770478A CN200710077047A CN101126688A CN 101126688 A CN101126688 A CN 101126688A CN A2007100770478 A CNA2007100770478 A CN A2007100770478A CN 200710077047 A CN200710077047 A CN 200710077047A CN 101126688 A CN101126688 A CN 101126688A
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Abstract
The utility model relates to a method for large-scale preparation of cell microscopic samples, which comprises the following steps under sterile conditions: cover slips with ferrous powders are arranged in the microscopic hole of a microplate, or a metal gasket attractable by magnetism is positioned at the bottom of the microscopic hole in advance, and then the ordinary cover slips are arranged on the metal gasket; single cell suspension is arranged on the cover slips and cultured; when the growth of the cell meets the requirements, the culture fluid is sucked out, and the cell is washed; the biochemical reaction is conducted in the microscopic hole; when the reaction is completed, the cell is washed again; a magnetically attractable board is flatly arranged for holding the cover slips, and stationary liquid for the cover slips is applied on the glass slide, then the microplate is inversely arranged on the glass slide, thus the cover slips are shifted on the glass slide under the influence of the magnetic force of the magnetic board. By adopting the cover slips with ferrous powders or arranging the metal gasket which can be adsorbed by the magnetism at the bottom of the cover slips, all the cover slips are sucked out and shifted on the glass slider via the magnetic board. The utility model has the advantages of convenient operation and high efficiency.
Description
Technical field
The present invention relates to a kind of method for preparing cell micro specimen, be meant a kind of method of large scale cell micro specimen preparation especially.
Background technology
The preparation cell micro specimen is life sciences such as biology, medical science and pharmacy experimental techniques commonly used.The method of the micro-sample of existing making cell is normally utilized such as cultivation vessel such as well plates earlier and is turned out cell, again cultured cell is built from cultivating in the vessel to take out to move on the microslide and with cover glass again, thereby finished the making of a cell micro specimen.There are distinct issues in the method for the micro-sample of existing this making cell: with cell when cultivating vessel and be transferred to the operating cost of microslide, inefficiency.
Summary of the invention
Technical matters to be solved by this invention is: provide a kind of method of large scale cell micro specimen preparation, its simple operation, high efficiency.
For solving the problems of the technologies described above, the present invention adopts following technical scheme: a kind of method of large scale cell micro specimen preparation, and it is included in the following steps of carrying out under the aseptic condition:
Place the cover glass step, the cover glass that will contain the iron powder composition is positioned in the micropore of microwell plate, and cover glass contacts with the micropore bottom comprehensively;
Cell culture step adds to the single cell suspension for preparing in advance on the cover glass by concrete requirement of experiment and to cultivate;
Clean the cell step, behind the cell growth certain hour, after the density of cell or state reached concrete requirement of experiment, the sucking-off nutrient solution cleaned cell again, and each shaking was cleaned 4-5 minute, inhaled and removed clean-out system;
The biochemical reaction step is carried out biochemical reaction in micropore, concrete reactions steps is carried out according to concrete experiment;
Clean the cell step once more, biochemical reaction cleans cell after finishing once more, and each shaking was cleaned 4-5 minute, to remove unnecessary reaction reagent, inhales after the cleaning again and removes clean-out system;
Take out the cover glass step, magnetic plate horizontal with surfacing with magnetic adsorptive power, magnetic plate area is consistent with the microwell plate area, again microslide numbering, one one are layered on above the magnetic plate, and on microslide coating cover glass immobile liquid, then microwell plate is inverted on the microslide, cover glass promptly is displaced downwardly on the microslide at the magneticaction of magnetic plate.
Further improvement in the technical proposal is: in the cell culture step, adopt 96 orifice plates, the single cell suspension addition is 200 microlitres/hole.
Further improvement in the technical proposal is: cleaning step and once more in the cleaning step, the clean-out system of employing is HBSS or PBS.
Further improvement in the technical proposal is: cleaning step reaches once more in the cleaning step, utilizes clean-out system to clean cell 3 times.
The utility model also provides following a kind of technical scheme: a kind of method of large scale cell micro specimen preparation, and it is included in the following steps of carrying out under the aseptic condition:
Place the cover glass step, earlier will with the cover glass micropore bottom that can be positioned over microwell plate of the same size by the metallic gasket that magnetic is adsorbed in, again the common lid slide is positioned on the metallic gasket, make and be close to tight between the two;
Cell culture step adds to the single cell suspension for preparing in advance on the cover glass by concrete requirement of experiment and to cultivate;
Clean the cell step, behind the cell growth certain hour, after the density of cell or state reached concrete requirement of experiment, the sucking-off nutrient solution cleaned cell again, and each shaking was cleaned 4-5 minute, inhaled and removed clean-out system;
The biochemical reaction step is carried out biochemical reaction in micropore, concrete reactions steps is carried out according to concrete experiment;
Clean the cell step once more, biochemical reaction cleans cell after finishing once more, and each shaking was cleaned 4-5 minute, to remove unnecessary reaction reagent, inhales after the cleaning again and removes clean-out system;
Take out the cover glass step, magnetic plate horizontal with surfacing with magnetic adsorptive power, magnetic plate area is consistent with the microwell plate area, again microslide numbering, one one are layered on above the magnetic plate, and on microslide coating cover glass immobile liquid, then microwell plate is inverted on the microslide, metallic gasket promptly promotes cover glass and moves on the microslide under the magneticaction of magnetic plate.
Further improvement in the technical proposal is: the stainless steel substrates of described metallic gasket for being adsorbed by magnetic.
The invention has the beneficial effects as follows: the cover glass that contains the iron powder composition by direct utilization places and comes cultured cell in the microwell plate; perhaps by cultured cell on the common lid slide that is lined with the metallic gasket that can be adsorbed by magnetic in the bottom surface; after cultured cells reaches requirement; and recycling magnetic plate with magnetic adsorptive power moves to the disposable whole sucking-offs of all cover glasses and makes cell micro specimen on the microslide; simple to operate; it is very convenient that the cover glass that especially will have a cell moves on the microslide; work efficiency is very high; can realize large scale cell micro specimen preparation; cooperate again and adopt 48 holes; the microwell plate in 96 holes comes cultured cell, and the slices that the efficiency ratio of its preparation sample is traditional are taken out the method for cover glass and wanted high more than 10 times.Method of the present invention can be widely used in subjects such as cell biology, pharmacy, medical science, science of heredity and carry out the cell micro specimen preparation.
Embodiment
In first embodiment of the invention, the invention provides a kind of method of large scale cell micro specimen preparation, it is included in the following steps of carrying out under the aseptic condition:
Place the cover glass step, the cover glass that will contain the iron powder composition is positioned in the micropore of microwell plate, and cover glass contacts with the micropore bottom comprehensively;
Cell culture step adds to the single cell suspension for preparing in advance on the cover glass by concrete requirement of experiment and to cultivate, and is example with the microwell plate that adopts 96 holes, and the addition of its single cell suspension is 200 microlitres/hole;
Clean the cell step, behind the cell growth certain hour, for example grow clone ball and need cultivate 10-20 days, after the density of cell or state reach concrete requirement of experiment, the sucking-off nutrient solution, clean cell again, each shaking was cleaned 4-5 minute, inhaled and removed clean-out system, preferably, the clean-out system that adopts is HBSS or PBS, generally cleans cell 3 times;
The biochemical reaction step is carried out biochemical reaction in micropore, for example antigen-antibody reaction, dyeing etc., and concrete reactions steps is carried out according to concrete experiment;
Clean the cell step once more, after biochemical reaction finishes, clean cell once more, each shaking was cleaned 4-5 minute, removing unnecessary reaction reagent, inhale again after the cleaning and remove clean-out system, and clean the cell step in the same manner, it is clean-out system that this step also preferably adopts HBSS or PBS, generally cleans cell 3 times;
Take out the cover glass step, the magnetic plate that will have magnetic attraction places the experiment desktop, magnetic plate area is consistent with the microwell plate area, again microslide numbering, one one are layered on above the magnetic plate, and on microslide coating cover glass immobile liquid, then microwell plate is inverted on the microslide, cover glass promptly is displaced downwardly on the microslide at the magneticaction of magnetic plate.
In the above-described first embodiment, be directly to have adopted the cover glass that contains the iron powder composition, so that when taking out cover glass, cover glass is drawn on the microslide by the magnetic plate, and in second embodiment of the invention, then carried out following variation: placing the cover glass step, do not adopt the cover glass that contains the iron powder composition, but earlier will with the cover glass micropore bottom that can be positioned over microwell plate of the same size by the metallic gasket that magnetic is adsorbed in, again the common lid slide is positioned on the metallic gasket, makes and be close to tight between the two; Like this, in taking out the cover glass step, when microwell plate was inverted on the microslide, this metallic gasket promptly can promote cover glass and move on the microslide under magneticaction.Thereby can easily cover glass be taken out in micropore and move to equally and form cell micro specimen on the microslide.
The cover glass that contains the iron powder composition by direct utilization places and comes cultured cell in the microwell plate; perhaps by cultured cell on the common lid slide that is lined with the metallic gasket that can be adsorbed by magnetic in the bottom surface; after cultured cells reaches requirement; and recycling magnetic plate with magnetic adsorptive power moves to the disposable whole sucking-offs of all cover glasses and makes cell micro specimen on the microslide; simple to operate; it is very convenient that the cover glass that especially will have a cell moves on the microslide; work efficiency is very high; can realize large scale cell micro specimen preparation; cooperate again and adopt 48 holes; the microwell plate in 96 holes comes cultured cell, and the slices that the efficiency ratio of its preparation sample is traditional are taken out the method for cover glass and wanted high more than 10 times.Method of the present invention can be widely used in subjects such as cell biology, pharmacy, medical science, science of heredity and carry out the cell micro specimen preparation.
Claims (9)
1. the method for a large scale cell micro specimen preparation is characterized in that, is included in the following steps of carrying out under the aseptic condition:
Place the cover glass step, the cover glass that will contain the iron powder composition is positioned in the micropore of microwell plate, and cover glass contacts with the micropore bottom comprehensively;
Cell culture step adds to the single cell suspension for preparing in advance on the cover glass by concrete requirement of experiment and to cultivate;
Clean the cell step, behind the cell growth certain hour, after the density of cell or state reached concrete requirement of experiment, the sucking-off nutrient solution cleaned cell again, and each shaking was cleaned 4-5 minute, inhaled and removed clean-out system;
The biochemical reaction step is carried out biochemical reaction in micropore, concrete reactions steps is carried out according to concrete experiment;
Clean the cell step once more, biochemical reaction cleans cell after finishing once more, and each shaking was cleaned 4-5 minute, to remove unnecessary reaction reagent, inhales after the cleaning again and removes clean-out system;
Take out the cover glass step, magnetic plate horizontal with surfacing with magnetic adsorptive power, magnetic plate area is consistent with the microwell plate area, again microslide numbering, one one are layered on above the magnetic plate, and on microslide coating cover glass immobile liquid, then microwell plate is inverted on the microslide, cover glass promptly is displaced downwardly on the microslide at the magneticaction of magnetic plate.
2. the method for a kind of large scale cell micro specimen preparation as claimed in claim 1, it is characterized in that: in the cell culture step, adopt 96 orifice plates, the single cell suspension addition is 200 microlitres/hole.
3. the method for a kind of large scale cell micro specimen preparation as claimed in claim 1 is characterized in that: cleaning step and once more in the cleaning step, the clean-out system of employing is HBSS or PBS.
4. as the method for claim 1 or 3 described a kind of large scale cell micro specimen preparations, it is characterized in that: cleaning step reaches once more in the cleaning step, utilizes clean-out system to clean cell 3 times.
5. the method for a large scale cell micro specimen preparation is characterized in that, is included in the following steps of carrying out under the aseptic condition:
Place the cover glass step, will with the cover glass micropore bottom that can be positioned over microwell plate earlier of the same size by the metallic gasket that magnetic is adsorbed in, again the common lid slide is positioned on the metallic gasket, make and be close to tight between the two;
Cell culture step adds to the single cell suspension for preparing in advance on the cover glass by concrete requirement of experiment and to cultivate;
Clean the cell step, behind the cell growth certain hour, after the density of cell or state reached concrete requirement of experiment, the sucking-off nutrient solution cleaned cell again, and each shaking was cleaned 4-5 minute, inhaled and removed clean-out system;
The biochemical reaction step is carried out biochemical reaction in micropore, concrete reactions steps is carried out according to concrete experiment;
Clean the cell step once more, biochemical reaction cleans cell after finishing once more, and each shaking was cleaned 4-5 minute, to remove unnecessary reaction reagent, inhales after the cleaning again and removes clean-out system;
Take out the cover glass step, magnetic plate horizontal with surfacing with magnetic adsorptive power, magnetic plate area is consistent with the microwell plate area, again microslide numbering, one one are layered on above the magnetic plate, and on microslide coating cover glass immobile liquid, then microwell plate is inverted on the microslide, metallic gasket promptly promotes cover glass and moves on the microslide under the magneticaction of magnetic plate.
6. the method for a kind of large scale cell micro specimen preparation as claimed in claim 5, it is characterized in that: in the cell culture step, adopt 96 orifice plates, the single cell suspension addition is 200 microlitres/hole.
7. the method for a kind of large scale cell micro specimen preparation as claimed in claim 5 is characterized in that: cleaning step and once more in the cleaning step, the clean-out system of employing is HBSS or PBS.
8. as the method for claim 5 or 7 described a kind of large scale cell micro specimen preparations, it is characterized in that: cleaning step reaches once more in the cleaning step, utilizes clean-out system to clean cell 3 times.
9. the method for a kind of large scale cell micro specimen preparation as claimed in claim 5 is characterized in that: the stainless steel substrates of described metallic gasket for being adsorbed by magnetic.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710077047A CN101126688B (en) | 2007-09-11 | 2007-09-11 | Large scale cell micro specimen preparation method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710077047A CN101126688B (en) | 2007-09-11 | 2007-09-11 | Large scale cell micro specimen preparation method |
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| CN101126688A true CN101126688A (en) | 2008-02-20 |
| CN101126688B CN101126688B (en) | 2010-05-26 |
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| CN200710077047A Expired - Fee Related CN101126688B (en) | 2007-09-11 | 2007-09-11 | Large scale cell micro specimen preparation method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109001003A (en) * | 2018-06-22 | 2018-12-14 | 中国科学院长春应用化学研究所 | For freezing the preparation method of electron microscope observation cell membrane sample |
| WO2023065299A1 (en) * | 2021-10-22 | 2023-04-27 | 深圳先进技术研究院 | Microscopic sample preparation device and cell phenotype control device |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100837265B1 (en) * | 2000-12-22 | 2008-06-11 | 킬레유니버시티 | A method of mechanically stimulating tissue forming cells for tissue engineering and regeneration applications |
| CN1869205A (en) * | 2006-05-23 | 2006-11-29 | 浙江大学 | Preparation method of immortalization pig liver cell |
| CN100470296C (en) * | 2007-08-28 | 2009-03-18 | 深圳职业技术学院 | Microscopic glass and its fetching device |
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2007
- 2007-09-11 CN CN200710077047A patent/CN101126688B/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109001003A (en) * | 2018-06-22 | 2018-12-14 | 中国科学院长春应用化学研究所 | For freezing the preparation method of electron microscope observation cell membrane sample |
| WO2023065299A1 (en) * | 2021-10-22 | 2023-04-27 | 深圳先进技术研究院 | Microscopic sample preparation device and cell phenotype control device |
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| CN101126688B (en) | 2010-05-26 |
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