CN103952291B - A kind of device of Cell migration assay - Google Patents
A kind of device of Cell migration assay Download PDFInfo
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- CN103952291B CN103952291B CN201410206458.2A CN201410206458A CN103952291B CN 103952291 B CN103952291 B CN 103952291B CN 201410206458 A CN201410206458 A CN 201410206458A CN 103952291 B CN103952291 B CN 103952291B
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- hole
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- cell migration
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
- B01L3/50855—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates using modular assemblies of strips or of individual wells
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A kind of device of Cell migration assay, relate to a kind of use for laboratory utensil, comprise plate body and be evenly distributed on the hole on plate body, hole is set to square, outside bottom hole is provided with grid, the medullary ray of grid overlaps with the axis in hole, and detached strips is posted in the inner side bottom hole, and the material of bonding is medical polymer tackiness agent, detached strips longitudinal cenlerline overlaps with the medullary ray of grid, the width of described detached strips is 1mm, and length is greater than bore length, is provided with holding part in one end of detached strips.The present invention is by being designed to square opening, and the area of cut is rectangle, and areal calculation is more accurate compared with circular port; By posting the detached strips of band draw ring in hole, when scratch experiment, detached strips can be taken off, the cut of neat specification in the same size can be obtained, and fast and easy; By being provided with grid bottom hole, conveniently can observing the position of statistics signaling, calculating relative scratch area.
Description
Technical field
The present invention relates to a kind of use for laboratory utensil, specifically a kind of device of Cell migration assay.
Background technology
Cell migration is ubiquity in the normal physiological activity of human body and disease occur, and the looking for food of such as cell, wound trauma healing, embryo's generation, neural system formation, immune response, cancer metastasis etc. much event all relate to cell migration.Cell migration is an important directions and the focus of current life scientific research, and scientists attempts the research by cell migration, in blood vessel injury reparation, medical use such as prevention cancer metastasis, skin-grafting etc., obtain larger achievement.
Current research cell migration mainly uses scratch experiment.Epithelial cell, keratinocyte etc. form individual layer or stratified epithelium under physiological status, and under the pathological states such as wound healing, cells migrate, based on the motion of side direction.Scratch experiment simulates this mode of motion well, and easy to operate, is therefore a good model of research cell migration.In existing scratch experiment, the aseptic plastics rifle head artificial hand of general employing is drawn, as shown in Figure 1, scratches visible edge is irregular, cell residual after having PBS to clean in cut, in different hole, scratch area is different, and these all can affect the judgement of experiment effect and result, and therefore this kind of method has the problems such as cut is not straight, scoring position is inaccurate, scratch area disunity.
Summary of the invention
The object of the present invention is to provide a kind of device of Cell migration assay, cut can be carried out easily and fast, and cut is neatly in the same size, bottom plate body, has grid scale marks, be convenient to observe and calculate relative scratch area.
The technical solution adopted for the present invention to solve the technical problems is: a kind of device of Cell migration assay, it is characterized in that, comprise plate body and be evenly distributed on the hole on plate body, hole is set to square, outside bottom hole is provided with grid, the medullary ray of grid overlaps with the axis in hole, detached strips is posted in inner side bottom hole, the material of bonding is medical polymer tackiness agent, detached strips longitudinal cenlerline overlaps with the medullary ray of grid, the width of described detached strips is 1mm, and length is greater than bore length, is provided with holding part in one end of detached strips.
Described grid distance is 0.25mm.
The holding part of described detached strips to be positioned at outside hole and not higher than plate lid, and end is provided with draw ring.
Described detached strips is material ultrathin transparent plastic glue strip.
The invention has the beneficial effects as follows: by being designed to square opening, the area of cut is rectangle, and areal calculation is more accurate compared with circular port; By posting the detached strips of band draw ring in hole, when scratch experiment, detached strips can be taken off, the cut of neat specification in the same size can be obtained, and fast and easy; By being provided with grid bottom hole, conveniently can observing the position of statistics signaling, calculating relative scratch area.
Accompanying drawing explanation
Fig. 1 is the rim condition schematic diagram in routine experimentation after artificial cut,
Fig. 2 is structural representation of the present invention,
Fig. 3 is bottom surface, hole upward view,
Fig. 4 is bottom surface, hole vertical view,
Fig. 5 is the A-A view of Fig. 4,
Fig. 6 is the rim condition schematic diagram after taking detached strips off.
In figure: 1 plate body, 2 holes, 3 detached stripss, 31 holding parts, 32 draw rings, 4 grids.
Embodiment
A device for Cell migration assay, the hole 2 comprising plate body 1 and be evenly distributed on plate body 1, hole 2 is set to square.Can determine the quantity in the hole needed according to demand, the outside bottom hole 2 is provided with grid 4, and grid distance is 0.25mm, and the medullary ray of grid 4 overlaps with the axis in hole 2.Detached strips 3 is posted for material ultrathin transparent plastic glue strip in inner side bottom hole 2, and the material of bonding is medical polymer tackiness agent, no cytotoxicity, removes noresidue bottom detached strips 3 metapore 2, does not affect Growth of Cells.Detached strips 3 longitudinal cenlerline overlaps with the medullary ray of grid 4, and like this after detached strips 3 is taken off, the cell of detached strips 3 both sides is identical relative to the distance of grid 4 medullary ray, is convenient to data statistics.The width of described detached strips 3 is 1mm, and length is greater than square opening length, rounded opening edge can be avoided like this to have part cell not separate and the scratch area caused calculates inaccurate problem.Be provided with holding part 31 in one end of detached strips 3, holding part is positioned at outside hole and not higher than plate lid, holding part 31 is provided with draw ring 32 higher than edge section, hole 2, is convenient to staff and takes tweezers clamping to make cut.
Experimental procedure:
1. cell cultures
For Bel7402 BEL-7402, tumour cell at RPMI1640 substratum (containing 10%FBS, 100U/ml penicillin, 100ug/ml Streptomycin sulphate), 37 DEG C, cultivate under 5%CO2 condition, when cell reaches 90% degrees of fusion, peptic cell counts.
(1) 75% alcohol wipe super clean bench table top, sets the plastic suction pipe required for operation, culturing bottle etc. successively, uses after uviolizing 30min; PBS phosphate buffered saline buffer, RPMI1640 complete culture solution and 0.25% pancreatin are placed in the pre-temperature of 37 DEG C of incubators, put into super clean bench after alcohol wipe;
(2) wear aseptic powder-free gloves, take out culturing bottle and be placed in observation of cell growth conditions under inverted phase contrast microscope, can count by peptic cell to during about 90% degrees of fusion until Growth of Cells;
(3) suction pipe is inhaled and is abandoned old nutrient solution, and with PBS wash buffer 2 times, add appropriate pancreatin, the amount of pancreatin, just to cover cell, is placed on if desired in 37 DEG C of incubators and hatches;
(4) basis of microscopic observation peptic cell, treats that kytoplasm bounces back, and intercellular substance increases, and visual inspection cell is sand-like coming off, and shows cell dissociation appropriateness, inhales and abandon Digestive system, adds complete culture solution and stops digestion;
(5) with suction pipe, the cell digested softly is blown and beaten into cell suspension.
2. cell counting and kind plate
(1) with 75% cotton ball soaked in alcohol by blood counting chamber and cover glass wiped clean, cover glass is covered on blood counting groove;
(2) by cell suspension piping and druming to be measured evenly, then drawing a small amount of suspension and slowly instill along cover glass edge, be full of suspension under ensureing cover glass, can not there is bubble in attention, suspension can not be allowed to flow in the groove of side, count after leaving standstill 3min;
(3) blood counting chamber is placed in microscope low power Microscopic observation, counts the total cellular score of 4 jiaos of 4 large lattice (each large lattice contain 16 middle lattice), press formulae discovery cell density below:
Cell count/the ml=(total cellular score/4 of 4 large lattice) × 10 of cell suspension
4;
3. scratch experiment
(1) by cell suspension respectively filling orifice 2, to cytogamy degree about 90% time test;
(2) clamp holding part 31 with aseptic nipper gently detached strips 3 to be taken off, owing to adopting medical polymer tackiness agent, no cytotoxicity, remove noresidue bottom detached strips 3 metapore 2, do not affect Growth of Cells;
(3) wash cell 3 times with PBS, add serum free medium, choose experimental point and Taking Pictures recording under microscope, 37 DEG C, 5%CO2 incubator is cultivated;
Observe after (4) 48 hours, judge cell position by grid 4, thus judge different group travelling speed, and take pictures;
(5) Image-Pro Plus6.0 software is used to analyze.Calculate relative scratch area.Each group of scratch area all with BEL7402 cell 0h scratch area for standard gets relative value, relative scratch area=48h scratch area/0h scratch area.
Claims (4)
1. the device of a Cell migration assay, it is characterized in that, comprise plate body and be evenly distributed on the hole on plate body, hole is set to square, outside bottom hole is provided with grid, the medullary ray of grid overlaps with the axis in hole, and detached strips is posted in the inner side bottom hole, and the material of bonding is medical polymer tackiness agent, detached strips longitudinal cenlerline overlaps with the medullary ray of grid, described detached strips width is 1mm, and length is greater than bore length, is provided with holding part in one end of detached strips.
2. the device of a kind of Cell migration assay according to claim 1, is characterized in that, described grid distance is 0.25mm.
3. the device of a kind of Cell migration assay according to claim 1 and 2, is characterized in that, the holding part of described detached strips to be positioned at outside hole and not higher than plate lid, and end is provided with draw ring.
4. the device of a kind of Cell migration assay according to the arbitrary claim of claim 1 or 2, is characterized in that, described detached strips is material ultrathin transparent plastic glue strip.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410206458.2A CN103952291B (en) | 2014-05-15 | 2014-05-15 | A kind of device of Cell migration assay |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410206458.2A CN103952291B (en) | 2014-05-15 | 2014-05-15 | A kind of device of Cell migration assay |
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| Publication Number | Publication Date |
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| CN103952291A CN103952291A (en) | 2014-07-30 |
| CN103952291B true CN103952291B (en) | 2015-08-05 |
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| CN106520519B (en) * | 2016-11-11 | 2018-11-27 | 四川大学华西医院 | One kind testing scale scoring devices for Wound healing cells |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN2830408Y (en) * | 2005-07-20 | 2006-10-25 | 韩露 | Cell culture plate |
| KR20100117869A (en) * | 2009-04-27 | 2010-11-04 | 한국산업기술대학교산학협력단 | Cell culture container |
| GB201010736D0 (en) * | 2010-06-25 | 2010-08-11 | Imp Innovations Ltd | IWAP (Interwell assay plate) |
| CN203820769U (en) * | 2014-05-15 | 2014-09-10 | 山东省千佛山医院 | Device for cell migration experiments |
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