CN102533537A - Device and method for screening growth and migration behaviors of cells in three-dimensional matrixes at high throughput - Google Patents

Device and method for screening growth and migration behaviors of cells in three-dimensional matrixes at high throughput Download PDF

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CN102533537A
CN102533537A CN2011104490415A CN201110449041A CN102533537A CN 102533537 A CN102533537 A CN 102533537A CN 2011104490415 A CN2011104490415 A CN 2011104490415A CN 201110449041 A CN201110449041 A CN 201110449041A CN 102533537 A CN102533537 A CN 102533537A
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dimensional
screening
cell
behaviors
growth
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CN102533537B (en
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赵三军
赵敏
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Yunnan Normal University
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Yunnan Normal University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/14—Scaffolds; Matrices
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/04—Flat or tray type, drawers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/38—Caps; Covers; Plugs; Pouring means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability

Abstract

The invention discloses a device and a method for screening growth and migration behaviors of cells in three-dimensional matrixes at high throughput, which relate to the field of biology. An objective is arranged below an electric objective table; a culture room plate is arranged above the electric objective table; a glass slide is arranged at the bottom part in the culture room plate; a polydimethylsiloxane thin layer covers on the glass slide; a cell suspension layer is coated onto the polydimethylsiloxane thin layer; both sides of the cell suspension layer are respectively provided with an adhesive tape; and cover glass covers on the adhesive tapes. According to the device and the method for screening the growth and migration behaviors of the cells in the three-dimensional matrixes at the high throughput, the defect that the information of important behaviors of the cells under a three-dimensional state, such as growth and migration, cannot be provided by two-dimensional research is overcome. The invention provides a novel high-throughput three-dimensional screening device and a novel high-throughput three-dimensional screening method. The device not only is capable of monitoring various behaviors of the cells under the three-dimensional state in real time but also is capable of conveniently and efficiently screening the influences of various matrixes on the behaviors of the cells at the high throughput while screening the influences of different matrixes and different factors on the important behaviors of the cells under the three-dimensional state at the high throughput, such as the growth and the migration. Moreover, the device has low price and a simple structure.

Description

In three dimensional matrix, the grow device and method of migratory behaviour of a kind of high flux screening cell
Technical field
The present invention relates to field of biology, be specifically related in three dimensional matrix, the grow device and method of migratory behaviour of a kind of high flux screening cell.
Background technology
The growth of cell and migration are the important steps of numerous important cells biological procedureses such as ontogeny, tissue repair, neomorph, the pernicious transfer of cancer, and cell has very big-difference in behaviors such as the growth of two dimension and migration and three-dimensional state.At present, mostly the growth of research cell and the device of migration are on the two dimensional surface device, to carry out, and research shows, produces polarization in the three-dimensional state cultured cells, and form the botryoidalis clone, but but do not have this characteristic in the two-dimensional state cultured cells.In addition, cell state in vivo all is to exist with three dimensional form, and extracellular various matrix and environment all can influence behaviors such as growth and the migration of cell.Therefore cell more approaches cell state in vivo in the growth of three-dimensional state and migration etc.Still the device that does not have at present behavior such as cell growth and migration under a kind of high flux screening three-dimensional state.
Summary of the invention
The purpose of this invention is to provide in three dimensional matrix, the grow device and method of migratory behaviour of a kind of high flux screening cell; It has overcome existing two-dimentional research can't provide the deficiency of cell in important behavioural informations such as three-dimensional state growth and migrations; A kind of high-throughput three-dimensional screening new device and method are provided; This device is not only monitored the various actions of cell at three-dimensional state in real time; And can high flux screening different substrates, different factor pair cell in important behaviors such as the growth of three-dimensional state and migration, the influence of convenient, efficient, high-throughout screening various kinds of substrates pair cell behavior, and this device is cheap, simple in structure.
In order to solve the existing problem of background technology; The present invention takes following technical scheme: its screening plant comprises slide glass 1, YSR 3286 thin layer 2, deckglass 3, adhesive tape 4, object lens 5, motorized subject table 6 and culturing room's plate 7; The below of motorized subject table 6 is provided with object lens 5, and the top of motorized subject table 6 is provided with culturing room's plate 7, and culturing room's plate 7 inner bottom parts are provided with slide glass 1; Be coated with YSR 3286 thin layer 2 on the slide glass 1; Scribble the cell suspension liquid layer on the YSR 3286 thin layer 2, cell suspension liquid layer both sides are provided with adhesive tape 4, are coated with deckglass 3 on the adhesive tape 4.
Its screening method is: (1) covers slide surface and in stink cupboard, leaves standstill the formation hydrophobic interfaces with YSR 3286 earlier, and (concentration is 10 to get 50 μ l cell suspensions 5Cell ml 1) and equal-volume matrix mixing;
(2) get 10-20 μ l mixed solution with micropipet; Cell mixture is evenly put to slide specimen viewing area formation matrix successively, is example with 5 * 5 matrixes, and viewing area is 1.5cm; Sample diameter 1.2mm; Sample interval is about 3mm, and the single sample height is about 0.5mm, forms the sample matrix that quantity does not wait thereby also can observe the district according to institute's requirement body customization sample;
(3) matrix is left standstill condense; Covered; Place the microscopically that is equipped with motorized subject table then, through motorized subject table and each matrix dot position of object lens definition, through automatic photography system interval acquiring image at regular intervals; Thereby obtain states such as the growth of cell in various three dimensional matrixes, migration, and the influence of behaviors such as the various extracellular matrix cell growth of high flux screening, migration.
The present invention has following beneficial effect: it has overcome existing two-dimentional research can't provide the deficiency of cell in important behavioural informations such as three-dimensional state growth and migrations; A kind of high-throughput three-dimensional screening new device and method are provided; This device is not only monitored the various actions of cell at three-dimensional state in real time; And can high flux screening different substrates, different factor pair cell in important behaviors such as the growth of three-dimensional state and migration; The influence of convenient, efficient, high-throughout screening various kinds of substrates pair cell behavior, and this device is cheap, simple in structure.
Description of drawings:
Fig. 1 is a structural representation of the present invention;
Fig. 2 is the structural representation of slide glass among the present invention.
Embodiment:
With reference to Fig. 1-2; This embodiment is taked following technical scheme: its screening plant comprises slide glass 1, YSR 3286 thin layer 2, deckglass 3, adhesive tape 4, object lens 5, motorized subject table 6 and culturing room's plate 7; The below of motorized subject table 6 is provided with object lens 5, and the top of motorized subject table 6 is provided with culturing room's plate 7, and culturing room's plate 7 inner bottom parts are provided with slide glass 1; Be coated with YSR 3286 thin layer 2 on the slide glass 1; Scribble the cell suspension liquid layer on the YSR 3286 thin layer 2, cell suspension liquid layer both sides are provided with adhesive tape 4, are coated with deckglass 3 on the adhesive tape 4.
Its screening method is: (1) covers slide surface and in stink cupboard, leaves standstill the formation hydrophobic interfaces with YSR 3286 earlier, gets 50 μ l cell suspension (concentration about 10 5Cell ml 1) and equal-volume matrix mixing;
(2) get 10-20 μ l mixed solution with micropipet; Cell mixture is evenly put to slide specimen viewing area formation matrix successively, is example with 5 * 5 matrixes, and viewing area is 1.5cm; Sample diameter 1.2mm; Sample interval is about 3mm, and the single sample height is about 0.5mm, forms the sample matrix that quantity does not wait thereby also can observe the district according to institute's requirement body customization sample;
(3) matrix is left standstill condense; Covered; Place the microscopically that is equipped with motorized subject table then, through motorized subject table and each matrix dot position of object lens definition, through automatic photography system interval acquiring image at regular intervals; Thereby obtain states such as the growth of cell in various three dimensional matrixes, migration, and the influence of behaviors such as the various extracellular matrix cell growth of high flux screening, migration.
This embodiment has overcome existing two-dimentional research can't provide the deficiency of cell in important behavioural informations such as three-dimensional state growth and migrations; A kind of high-throughput three-dimensional screening new device and method are provided; This device is not only monitored the various actions of cell at three-dimensional state in real time; And can high flux screening different substrates, different factor pair cell in important behaviors such as the growth of three-dimensional state and migration; The influence of convenient, efficient, high-throughout screening various kinds of substrates pair cell behavior, and this device is cheap, simple in structure.

Claims (2)

1. the high flux screening cell device and method of migratory behaviour of in three dimensional matrix, growing; The screening plant that it is characterized in that it comprises slide glass (1), YSR 3286 thin layer (2), deckglass (3), adhesive tape (4), object lens (5), motorized subject table (6) and culturing room's plate (7); The below of motorized subject table (6) is provided with object lens (5); The top of motorized subject table (6) is provided with culturing room's plate (7); Culturing room's plate (7) inner bottom part is provided with slide glass (1), is coated with YSR 3286 thin layer (2) on the slide glass (1), and the YSR 3286 thin layer scribbles the cell suspension liquid layer on (2); Cell suspension liquid layer both sides are provided with adhesive tape (4), and adhesive tape (is coated with deckglass (3) on 4.
2. In three dimensional matrix, the grow device and method of migratory behaviour of a kind of high flux screening cell according to claim 1; The screening method that it is characterized in that it is: (1) covers slide surface and in stink cupboard, leaves standstill the formation hydrophobic interfaces with YSR 3286 earlier, and getting concentration is 10 5 Cell ml 1 , the cell suspension of 50 μ l and equal-volume matrix mixing;
(3) are left standstill matrix and are condensed; Covered; Place the microscopically that is equipped with motorized subject table then, through motorized subject table and each matrix dot position of object lens definition, through automatic photography system interval acquiring image at regular intervals; Thereby obtain growth, the transition state of cell in various three dimensional matrixes, and the influence of behaviors such as the various extracellular matrix cell growth of high flux screening, migration.
CN 201110449041 2011-12-29 2011-12-29 Device and method for screening growth and migration behaviors of cells in three-dimensional matrixes at high throughput Expired - Fee Related CN102533537B (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111812095A (en) * 2020-09-08 2020-10-23 东南大学苏州医疗器械研究院 Photonic crystal microscope and cell mechanics measuring method
CN112391277A (en) * 2020-11-24 2021-02-23 北京科技大学 Experimental device and method for visual research on microbial diffusion and migration

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JP2005199394A (en) * 2004-01-16 2005-07-28 Aida Eng Ltd Adhesion method of pdms substrate and the other synthetic resin substrate, and method for manufacturing microchip
CN101245311A (en) * 2008-02-26 2008-08-20 武汉大学 Three-dimensional high-flux medicaments sifting chip and manufacture method thereof
CN101263223A (en) * 2005-09-16 2008-09-10 通用电气医疗集团生物科学公司 Cell migration assay
WO2011035177A2 (en) * 2009-09-17 2011-03-24 The Brigham And Women's Hospital, Inc. High-throughput screening
CN102197129A (en) * 2008-10-24 2011-09-21 可乐丽股份有限公司 Cell culture kit, screening method, and cell culture kit manufacturing method
CN102212458A (en) * 2011-03-31 2011-10-12 西北工业大学 Variable pitch microcolumn array-based cell sorting structure and manufacturing method thereof
CN102212459A (en) * 2011-03-31 2011-10-12 西北工业大学 Polydimethylsiloxane (PDMS)-based cell sorting structure

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005199394A (en) * 2004-01-16 2005-07-28 Aida Eng Ltd Adhesion method of pdms substrate and the other synthetic resin substrate, and method for manufacturing microchip
CN101263223A (en) * 2005-09-16 2008-09-10 通用电气医疗集团生物科学公司 Cell migration assay
CN101245311A (en) * 2008-02-26 2008-08-20 武汉大学 Three-dimensional high-flux medicaments sifting chip and manufacture method thereof
CN102197129A (en) * 2008-10-24 2011-09-21 可乐丽股份有限公司 Cell culture kit, screening method, and cell culture kit manufacturing method
WO2011035177A2 (en) * 2009-09-17 2011-03-24 The Brigham And Women's Hospital, Inc. High-throughput screening
CN102212458A (en) * 2011-03-31 2011-10-12 西北工业大学 Variable pitch microcolumn array-based cell sorting structure and manufacturing method thereof
CN102212459A (en) * 2011-03-31 2011-10-12 西北工业大学 Polydimethylsiloxane (PDMS)-based cell sorting structure

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111812095A (en) * 2020-09-08 2020-10-23 东南大学苏州医疗器械研究院 Photonic crystal microscope and cell mechanics measuring method
CN111812095B (en) * 2020-09-08 2020-12-25 东南大学苏州医疗器械研究院 Photonic crystal microscope and cell mechanics measuring method
WO2022052361A1 (en) * 2020-09-08 2022-03-17 东南大学苏州医疗器械研究院 Photonic crystal microscope and cell mechanics measurement method
CN112391277A (en) * 2020-11-24 2021-02-23 北京科技大学 Experimental device and method for visual research on microbial diffusion and migration

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