CN105624035A - Spiral-capillary-based magnetic cell sorting apparatus and method - Google Patents
Spiral-capillary-based magnetic cell sorting apparatus and method Download PDFInfo
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- CN105624035A CN105624035A CN201610084139.8A CN201610084139A CN105624035A CN 105624035 A CN105624035 A CN 105624035A CN 201610084139 A CN201610084139 A CN 201610084139A CN 105624035 A CN105624035 A CN 105624035A
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- capillary tube
- cell
- helical form
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- form capillary
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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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
Abstract
The invention discloses a spiral-capillary-based magnetic cell sorting apparatus and method, comprising four steps: S1, feeding a sample; S2, separating in a stationary manner; S3, washing at low flow speed; S4, collecting at high flow speed. A spiral capillary placed in the center of a circular magnet is used such that all cell and magnetic bead turbid liquids are near a tube wall of the circular magnet. As the tube wall of the circular magnet has highest magnetic field strength, it is possible to make full use of a magnetic field, improve sorting efficiency and reduce sorting time by using the apparatus. After step S4 is over, the spiral capillary is fully filled with a buffer solution, it is possible to continuously carry out the step S1 of sample feeding and start a next sorting procedure such that cells are continuously sorted; the invention is applicable to sorting of a large quantity of cells.
Description
Technical field
The invention belongs to biological technical field, particularly to a kind of Cell magnetic sorting unit based on helical form capillary tube and Cell magnetic sorting unit method.
Background technology
Cell magnetic sorting technology is made by being coupled on magnetic particle the identification of antibody high specific labelling targeted cell surface antigen, the mode of the strength screening magnetic bead in recycling magnetic field, the cell being not associated with magnetic bead is removed, obtain the target cell of purification, be the common technology of a kind of cell sorting and enrichment. The Cell magnetic sorted product being widely used at present mainly has two kinds, and one is magnetic separation frame, the EasySep of representative products such as StemCell companyTMMagnet, this is the strong magnets of a kind of customization, it is fitted in magnet surface by being contained in test tube by the cell suspension being marked with magnetic bead, then static a period of time, the cell being combined with magnetic bead will be attracted on the sidewall of Magnet, afterwards by the mode toppled over or utilize pipettor to remove clear liquid, what stay be exactly the target cell needing to sort. Another kind is sorting post, representative products such as .Miltenyi companySorting post, sorting post is the plastic containers that a class is filled with the iron shot of different size. Iron shot can play the effect that magnetic force expands in magnetic field, and there is hydrophilic being coated on surface simultaneously, will not damaging cells.
Sorting post is disposable products, need to not stop to change during continuous sorting, and cost is too high. Magnetic separation frame does not have the life-span to limit, but sorting also must change sorting test tube every time, it is impossible to realize continuously uninterrupted sorting. Therefore, this area does not need to change consumptive material in the urgent need to developing one, it is possible to achieve the cell sorting techniques of continuous sorting, meets again the requirement that the sorting time is short, efficiency is high simultaneously.
Summary of the invention
It is an object of the invention to provide a kind of Cell magnetic sorting unit based on helical form capillary tube and Cell magnetic sorting unit method, by adopting the helical form capillary tube being placed on annular magnet central authorities, the suspension of all cells and magnetic bead is allowed to be at the near-wall that annular magnet magnetic is the strongest. Then pass through adjustment liquid and flow through the flow velocity of capillary tube, it is possible to achieve the continuous sorting of cell.
In order to solve above-mentioned technical problem, the technical solution used in the present invention is: a kind of Cell magnetic sorting unit based on helical form capillary tube, including helical form capillary tube and ringshaped magnet, helical form capillary tube is placed on annular magnet central authorities, and it is close to the inwall of ringshaped magnet, the magnetic line of force direction of annular magnet is along the directional spreding being perpendicular to axis in circular hole, and helical form capillary tube has two ports, for filling opening and leakage fluid dram.
Described filling opening and leakage fluid dram are positioned at the homonymy of ringshaped magnet.
The Cell magnetic method for separating of the described Cell magnetic sorting unit based on helical form capillary tube, comprises the steps:
S1: sample is put into: the dirty solution including target cell, labelling magnetic bead and spuious cell is added from the filling opening of helical form capillary tube so that it is be full of helical form capillary tube;
S2: static separation: filling opening and leakage fluid dram are closed, and keeps a period of time, make firmly to be adsorbed on the tube wall of capillary tube under the influence of a magnetic field by the target cell of marked by magnetic bead;
S3: slug flow speed is cleaned: add cleanout fluid from filling opening so that it is slowly flow across capillary tube, will do not discharged by the spuious cell rearrangement liquid mouth of marked by magnetic bead;
S4: fast flow velocity is collected: add buffer from filling opening so that it is flow fast through capillary tube, will be washed away from tube wall by the target cell of marked by magnetic bead, discharges from leakage fluid dram, and is collected.
Described step S2 keep a period of time to refer to 2 minutes.
Described step S3 slowly flows across and refers to that flow velocity is 1mm/s.
Described step S4 flows fast through and refers to that flow velocity is 50mm/s.
The invention has the beneficial effects as follows:
The present invention is placed on the helical form capillary tube of annular magnet central authorities by adopting, and allows the suspension of all cells and magnetic bead be at the near-wall of annular magnet. Owing to annular magnet centre magnetic field intensity is very weak, tube wall place magnetic field intensity is the highest, adopts this device, it is possible to make full use of magnetic field, improves the efficiency of separation, reduces the sorting time. After S4 step terminates, in helical form capillary tube, being full of buffer, it is possible to proceed S1 sample and put into step, starting next one sorting flow process, thus realizing the continuous sorting of cell, it is adaptable to need a fairly large number of occasion of sorting cells.
Accompanying drawing explanation
Fig. 1 is the structural representation of cell magnetic force sorting device of the present invention.
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail, to separate CD3+ cell:
As it is shown in figure 1, the present invention based on cell magnetic force sorting device include: helical form capillary tube 3 and ringshaped magnet 4. Helical form capillary tube 3 caliber external diameter 1.5mm, internal diameter 1.12mm, helical form race diameter 17.8mm. There are two ports, for filling opening 1 and leakage fluid dram 2. Described filling opening and leakage fluid dram are positioned at the upside of ringshaped magnet. The material of annular magnet is Ru-Fe-Mn, is sized to internal diameter 18mm, external diameter 55mm, height 60mm. The magnetic line of force direction of annular magnet is along the directional spreding being perpendicular to axis in circular hole, and the closer to the place of inner wall surface, magnetic field intensity is more big. Helical form capillary tube 3 is placed on the central authorities of annular magnet 4, and is close to the inwall of ringshaped magnet. Magnetic field can be made full use of, improve the efficiency of separation, reduce the sorting time.
The Cell magnetic method for separating of the present invention, comprises the following steps:
S1: sample is put into; The dirty solution including target cell, labelling magnetic bead and spuious cell is added from the filling opening 1 of helical form capillary tube so that it is be full of helical form capillary tube;
S2: static separation; Filling opening 1 and leakage fluid dram 2 are closed, and keeps a period of time, make firmly to be adsorbed on the tube wall of capillary tube under the influence of a magnetic field by the target cell of marked by magnetic bead; Retention time in the present embodiment is 2 minutes.
S3: slug flow speed is cleaned; Add cleanout fluid from filling opening 1 so that it is slowly flow across capillary tube, will may not the spuious cell rearrangement liquid mouth 2 of marked by magnetic bead discharge; In the present embodiment, the flow velocity of cleanout fluid is 1mm/s, under this flow velocity, is attracted on the tube wall of capillary tube by the target cell of marked by magnetic bead, remains stationary as; And spuious cell flows along with liquid, flowing out from leakage fluid dram 2, this step can wash the spuious cell in dirty solution.
S4: fast flow velocity is collected; Add buffer from filling opening 1 so that it is flow fast through capillary tube, will be washed away from tube wall by the target cell of marked by magnetic bead, discharge from leakage fluid dram 2, and be collected. In the present embodiment, the flow velocity of cleanout fluid is 50mm/s;
After S4 step terminates, in helical form capillary tube, being full of buffer, it is possible to proceed S1 sample and put into step, starting next one sorting flow process, thus realizing the continuous sorting of cell, it is adaptable to need a fairly large number of occasion of sorting cells.
Embodiment described above is merely to illustrate technological thought and the feature of the present invention, its object is to make those skilled in the art it will be appreciated that present disclosure implementing according to this, the scope of the claims of the present invention only can not be limited with the present embodiment, what namely all disclosed spirit was made changes on an equal basis or modifies, and still drops in the scope of the claims of the present invention.
Claims (6)
1. the Cell magnetic sorting unit based on helical form capillary tube, it is characterized in that, including helical form capillary tube (3) and ringshaped magnet (4), helical form capillary tube (3) is placed on the central authorities of annular magnet (4), and it is close to the inwall of ringshaped magnet, the magnetic line of force direction of annular magnet is along the directional spreding being perpendicular to axis in circular hole, and helical form capillary tube has two ports, for filling opening (1) and leakage fluid dram (2).
2. the Cell magnetic sorting unit based on helical form capillary tube according to claim 1, it is characterised in that described filling opening (1) and leakage fluid dram (2) are positioned at the homonymy of ringshaped magnet (4).
3. the Cell magnetic method for separating of the Cell magnetic sorting unit based on helical form capillary tube as claimed in claim 1, it is characterised in that comprise the steps:
S1: sample is put into: the dirty solution including target cell, labelling magnetic bead and spuious cell is added from the filling opening (1) of helical form capillary tube so that it is be full of helical form capillary tube;
S2: static separation: filling opening (1) and leakage fluid dram (2) are closed, and keeps a period of time, make firmly to be adsorbed on the tube wall of capillary tube under the influence of a magnetic field by the target cell of marked by magnetic bead;
S3: slug flow speed is cleaned: add cleanout fluid from filling opening (1) so that it is slowly flow across capillary tube, will do not discharged by spuious cell rearrangement liquid mouth (2) of marked by magnetic bead;
S4: fast flow velocity is collected: add buffer from filling opening (1) so that it is flow fast through capillary tube, will be washed away from tube wall by the target cell of marked by magnetic bead, discharges from leakage fluid dram (2), and is collected.
4. the Cell magnetic method for separating based on helical form capillary tube according to claim 3, it is characterised in that keep a period of time to refer to 2 minutes in described step S2.
5. the Cell magnetic method for separating based on helical form capillary tube according to claim 1, it is characterised in that slowly flow across in described step S3 and refer to that flow velocity is 1mm/s.
6. the Cell magnetic method for separating based on helical form capillary tube according to claim 1, it is characterised in that flow fast through in described step S4 and refer to that flow velocity is 50mm/s.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610084139.8A CN105624035A (en) | 2016-02-04 | 2016-02-04 | Spiral-capillary-based magnetic cell sorting apparatus and method |
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| CN201610084139.8A CN105624035A (en) | 2016-02-04 | 2016-02-04 | Spiral-capillary-based magnetic cell sorting apparatus and method |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107083330A (en) * | 2017-04-17 | 2017-08-22 | 新疆出入境检验检疫局检验检疫技术中心 | Nucleic acid nano magnetic bead separating device and separation method |
| CN107488582A (en) * | 2017-08-08 | 2017-12-19 | 上海交通大学 | Micro fluidic device |
| CN109486651A (en) * | 2018-10-08 | 2019-03-19 | 广东菲鹏生物有限公司 | Separator and its application, detection system, electrochemical detection system and cell sorting system |
| CN109536434A (en) * | 2017-09-21 | 2019-03-29 | 北京航空航天大学青岛研究院 | Cell screening method and system based on immunomagnetic beads label |
| CN110004106A (en) * | 2018-04-12 | 2019-07-12 | 苏州天际创新纳米技术有限公司 | A kind of many cells magnetic separating method and system |
| CN110736422A (en) * | 2019-09-12 | 2020-01-31 | 中国地质大学（武汉） | prefabricated magnetic field layout system and deformation state response method |
| CN111139182A (en) * | 2018-11-02 | 2020-05-12 | 青岛华大智造普惠科技有限公司 | Magnetic screening device, micro-droplet screening system and magnetic screening method of micro-droplets |
| CN112662613A (en) * | 2019-10-15 | 2021-04-16 | 深圳市红莓生物科技有限公司 | Magnetic cell separation method |
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| US5795470A (en) * | 1991-03-25 | 1998-08-18 | Immunivest Corporation | Magnetic separation apparatus |
| US6143577A (en) * | 1996-05-07 | 2000-11-07 | Biocom S.A. | Process and installations for separation of magnetic particles in a fluid for biological analysis, and application of said process |
| CN104928280A (en) * | 2015-06-26 | 2015-09-23 | 关节动力安达（天津）生物科技有限公司 | Rotary magnetic cell separation method |
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2016
- 2016-02-04 CN CN201610084139.8A patent/CN105624035A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5795470A (en) * | 1991-03-25 | 1998-08-18 | Immunivest Corporation | Magnetic separation apparatus |
| US6143577A (en) * | 1996-05-07 | 2000-11-07 | Biocom S.A. | Process and installations for separation of magnetic particles in a fluid for biological analysis, and application of said process |
| CN104928280A (en) * | 2015-06-26 | 2015-09-23 | 关节动力安达（天津）生物科技有限公司 | Rotary magnetic cell separation method |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107083330A (en) * | 2017-04-17 | 2017-08-22 | 新疆出入境检验检疫局检验检疫技术中心 | Nucleic acid nano magnetic bead separating device and separation method |
| CN107488582A (en) * | 2017-08-08 | 2017-12-19 | 上海交通大学 | Micro fluidic device |
| CN109536434A (en) * | 2017-09-21 | 2019-03-29 | 北京航空航天大学青岛研究院 | Cell screening method and system based on immunomagnetic beads label |
| CN109536434B (en) * | 2017-09-21 | 2022-04-29 | 北京航空航天大学青岛研究院 | Method and system for screening cells based on immunomagnetic bead labeling |
| CN110004106A (en) * | 2018-04-12 | 2019-07-12 | 苏州天际创新纳米技术有限公司 | A kind of many cells magnetic separating method and system |
| CN109486651A (en) * | 2018-10-08 | 2019-03-19 | 广东菲鹏生物有限公司 | Separator and its application, detection system, electrochemical detection system and cell sorting system |
| CN111139182A (en) * | 2018-11-02 | 2020-05-12 | 青岛华大智造普惠科技有限公司 | Magnetic screening device, micro-droplet screening system and magnetic screening method of micro-droplets |
| CN111139182B (en) * | 2018-11-02 | 2023-11-14 | 青岛华大智造科技有限责任公司 | Magnetic screening device, micro-droplet screening system and micro-droplet magnetic screening method |
| CN110736422A (en) * | 2019-09-12 | 2020-01-31 | 中国地质大学（武汉） | prefabricated magnetic field layout system and deformation state response method |
| CN112662613A (en) * | 2019-10-15 | 2021-04-16 | 深圳市红莓生物科技有限公司 | Magnetic cell separation method |
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Application publication date: 20160601 |