CN103952291A - Device for cell migration experiment - Google Patents
Device for cell migration experiment Download PDFInfo
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- CN103952291A CN103952291A CN201410206458.2A CN201410206458A CN103952291A CN 103952291 A CN103952291 A CN 103952291A CN 201410206458 A CN201410206458 A CN 201410206458A CN 103952291 A CN103952291 A CN 103952291A
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- holes
- grid
- cell migration
- hole
- dividing strips
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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)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a device for cell migration experiment and relates to an appliance for a laboratory. The device comprises a plate body and holes uniformly distributed on the plate body, wherein the holes are square; a grid is arranged outside the bottoms of the holes; a center line of the grid is superposed with central axes of the holes; dividing strips are pasted inside the bottoms of the holes; the pasting material is a medical high polymer binder; longitudinal centre lines of the dividing strips are superposed with the center line of the grid; the dividing strips are 1 mm wide, and width of the dividing strips is greater than length of the holes; a holding part is arranged at one end of each of the dividing strips. According to the device for cell migration experiment disclosed by the invention, square holes are designed, and the area of scratches is rectangular, so that area calculation is more precise in comparison with that of circular holes; dividing strips with pulling rings are adhered in the holes, so that the dividing strips can be uncovered to obtain scratches which are consist in size and neat in specification while scratch experiment is carried out; moreover, the device is convenient and quick; the grid is arranged on the bottoms of the holes, so that cell migration positions can be conveniently observed and counted to calculate 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 experiment use.
Background technology
Cell migration is ubiquity in the normal physiological activity of human body and disease occur, such as the looking for food of cell, wound wound healing, embryo's generations, neural system formation, immune response, cancer metastasis etc. much events all relate to cell migration.Cell migration is important directions and the focus of current life science, and scientists is attempted the research by cell migration, in blood vessel injury reparation, stop aspect the medical uses such as cancer metastasis, skin-grafting and obtaining larger achievement.
Scratch experiment is mainly used in research cell migration at present.Epithelial cell, keratinocyte etc. form individual layer or stratified epithelium under physiological status, and under the pathological states such as wound healing, cell moves, and take the motion of side direction as main.Scratch experiment has been simulated 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, residual cell after having PBS to clean in cut, in different holes, cut area is different, and these all can affect the judgement of experiment effect and result, so this kind of method has the problems such as cut is not straight, cut position is inaccurate, cut area disunity.
Summary of the invention
The object of the present invention is to provide a kind of device of cell migration experiment use, can carry out easily and fast cut, and cut is neatly in the same size, grid scale marks is arranged at plate body bottom, is convenient to observe and calculate relative cut area.
The technical solution adopted for the present invention to solve the technical problems is: a kind of device of cell migration experiment use, it is characterized in that, comprise plate body and be evenly distributed on the hole on plate body, hole is set to square, outside in bottom, hole is provided with grid, the medullary ray of grid overlaps with the axis in hole, detached strips is posted in inner side in bottom, hole, bonding material 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, in one end of detached strips, is provided with holding part.
Described grid distance is 0.25mm.
The holding part of described detached strips is 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, calculate more accurate with circular port phase specific area; By post the detached strips with draw ring in hole, can when scratch experiment, take detached strips off, can obtain the cut of neat specification in the same size, and fast and easy; By being provided with grid in bottom, hole, can conveniently observe the position of statistics signaling, calculate relative cut area.
Accompanying drawing explanation
Fig. 1 is the rim condition schematic diagram after artificial cut in common experiment,
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 of taking off after detached strips.
In figure: 1 plate body, 2 holes, 3 detached stripss, 31 holding parts, 32 draw rings, 4 grids.
Embodiment
A device for cell migration experiment use, comprises plate body 1 and is evenly distributed on the hole 2 on plate body 1, hole 2 is set to square.The quantity that can determine according to demand the hole needing, in hole, the outside of 2 bottoms 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.In hole, detached strips 3 is posted for material ultrathin transparent plastic glue strip in the inner side of 2 bottoms, and bonding material is medical polymer tackiness agent, and no cytotoxicity is removed detached strips 3 metapore 2 bottom noresidues, does not affect Growth of Cells.Detached strips 3 longitudinal cenlerlines overlap with the medullary ray of grid 4, and after detached strips 3 is taken off, the cell of detached strips 3 both sides is identical with respect to the distance of grid 4 medullary rays, is convenient to data statistics like this.The width of described detached strips 3 is 1mm, and length is greater than square opening length, can avoid like this circular port edge to have part cell not separate and the cut area that causes calculates inaccurate problem.In one end of detached strips 3, be provided with holding part 31, holding part is positioned at outside hole and not higher than plate lid, holding part 31 is provided with draw ring 32 higher than 2 edge sections, hole, is convenient to staff and takes tweezers clamping to make cut.
Experimental procedure:
1. cell cultures
Take Bel7402 BEL-7402 as example, tumour cell (contains 10%FBS at RPMI1640 substratum, 100U/ml penicillin, 100ug/ml Streptomycin sulphate), cultivate under 37 ℃, 5%CO2 condition, when cell reaches 90% degrees of fusion, peptic cell is counted.
(1) 75% alcohol wipe super clean bench table top, sets the needed plastic suction pipe of operation, culturing bottle etc. successively, after uviolizing 30min, uses; PBS phosphate buffered saline buffer, RPMI1640 complete culture solution and 0.25% pancreatin are placed in the pre-temperature of 37 ℃ 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, until Growth of Cells, can count by peptic cell during to about 90% degrees of fusion;
(3) suction pipe is inhaled and is abandoned old nutrient solution, with PBS damping fluid, rinses 2 times, adds appropriate pancreatin, and the amount of pancreatin, just to cover cell, is placed on if desired in 37 ℃ of incubators and hatches;
(4) micro-Microscopic observation peptic cell, treats kytoplasm retraction, and intercellular substance increases, and visual inspection cell is sand-like and comes off, and shows cell dissociation appropriateness, inhales and abandons Digestive system, adds complete culture solution to stop digestion;
(5) with suction pipe, the cell digesting is is softly 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 splash into along cover glass edge, guarantee to be full of suspension under cover glass, can not there is bubble in attention, can not allow suspension flow in the groove of side, after standing 3min, counts;
(3) blood counting chamber is placed in to microscope low power Microscopic observation, counts the total cellular score of 4 jiaos of 4 large lattice (each large lattice contains 16 middle lattice), press formula below and calculate cell density:
Cell count/the ml=of cell suspension (total cellular score/4 of 4 large lattice) * 10
4;
3. scratch experiment
(1), by cell suspension respectively in filling orifice 2, during to cytogamy degree approximately 90%, test;
(2) with aseptic nipper, clamp holding part 31 and gently detached strips 3 is taken off, owing to adopting medical polymer tackiness agent, no cytotoxicity, removes detached strips 3 metapore 2 bottom noresidues, does not affect Growth of Cells;
(3) with PBS, wash cell 3 times, add serum free medium, choose experimental point Taking Pictures recording under microscope, 37 ℃, 5%CO2 incubator is cultivated;
After (4) 48 hours, observe, by grid 4 judgement cell positions, thereby judge different group travelling speeies, and take pictures;
(5) use Image-Pro Plus6.0 software to analyze.Calculate relative cut area.Each is organized cut area and all take BEL7402 cell 0h cut area and get relative value as standard, relatively cut area=48h cut area/0h cut area.
Claims (4)
1. the device of cell migration experiment use, it is characterized in that, comprise plate body and be evenly distributed on the hole on plate body, hole is set to square, outside in bottom, hole is provided with grid, the medullary ray of grid overlaps with the axis in hole, in the inner side of bottom, hole, posts detached strips, and bonding material 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, in one end of detached strips, is provided with holding part.
2. the device of a kind of cell migration experiment use according to claim 1, is characterized in that, described grid distance is 0.25mm.
3. the device of a kind of cell migration experiment use according to claim 1 and 2, is characterized in that, the holding part of described detached strips is positioned at outside hole and not higher than plate lid, and end is provided with draw ring.
4. according to the device of a kind of cell migration experiment use described in the arbitrary claim of claim 1 or 2, it 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 |
Applications Claiming Priority (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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| Publication Number | Publication Date |
|---|---|
| CN103952291A true CN103952291A (en) | 2014-07-30 |
| CN103952291B CN103952291B (en) | 2015-08-05 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201410206458.2A Active CN103952291B (en) | 2014-05-15 | 2014-05-15 | A kind of device of Cell migration assay |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520519A (en) * | 2016-11-11 | 2017-03-22 | 四川大学华西医院 | Ruler scratching device used for wound healing cell experiments |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| WO2011161480A1 (en) * | 2010-06-25 | 2011-12-29 | Imperial Innovations Ltd | Multi-well assay plate |
| CN203820769U (en) * | 2014-05-15 | 2014-09-10 | 山东省千佛山医院 | Device for cell migration experiments |
-
2014
- 2014-05-15 CN CN201410206458.2A patent/CN103952291B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| WO2011161480A1 (en) * | 2010-06-25 | 2011-12-29 | Imperial Innovations Ltd | Multi-well assay plate |
| CN203820769U (en) * | 2014-05-15 | 2014-09-10 | 山东省千佛山医院 | Device for cell migration experiments |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520519A (en) * | 2016-11-11 | 2017-03-22 | 四川大学华西医院 | Ruler scratching device used for wound healing cell experiments |
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| CN103952291B (en) | 2015-08-05 |
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