CN104568539B - Automatic histocyte separator - Google Patents
Automatic histocyte separator Download PDFInfo
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- CN104568539B CN104568539B CN201410772430.5A CN201410772430A CN104568539B CN 104568539 B CN104568539 B CN 104568539B CN 201410772430 A CN201410772430 A CN 201410772430A CN 104568539 B CN104568539 B CN 104568539B
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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
- C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
- C12M3/08—Apparatus for tissue disaggregation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- Biotechnology (AREA)
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- Sampling And Sample Adjustment (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses an automatic histocyte separator which is used for separating tissues into single cells. The separator comprises a separation tube and a motor, wherein the separation tube comprises a tube wall and a tube core in the tube wall; the top part of the tube wall is blocked; a tube cover covers the bottom part of the tube wall; the tube core comprises a stator sleeve and a rotor in the sleeve; a rotor joint is arranged on the tube cover; the rotor is connected with the rotor joint; the motor is located outside the separation tube, and is used for providing rotary power for the rotor through the rotor joint; and the inside of the tube core is sequentially divided into an archimedes spiral pipe area, a cell separation area and a negative pressure rotary cutting area from top to bottom. An automatic system with a special tube core is utilized; a microprocessor is used for defining special application program to optimize operation regulations; a specimen can be conveniently treated; the cell separation and preparation processes are standardized and programmed; and the effects on human factors are reduced.
Description
Technical field
The present invention relates to cell separation, more particularly to a kind of histocyte automatic segregator.
Background technology
Histocyte is to carry out the life sciences such as cell biology, molecular biology, immunology, science of heredity, oncology to grind
The important materials studied carefully, for example, carry out flow cytometry（Flow cytometry, FCM）The premise of detection requires that sample is slender
Born of the same parents' suspension, and most of solid tissues are all difficult to make high-quality single cell suspension sample, thus FCM is greatly limit to reality
Body tissue is particularly the analysis and research of solid tumor.Histocyte separator is used for the body tissue such as dispersion such as lung, liver, tumour
For individual cells suspension, the tradition that solid tissue is separated into individual cells is prepared into strategy and typically adopts manual method, mainly had
Mechanical shearing method, enzyme digestion, detergent method of chemical treatment, syringe needle suction method etc..
In order to the automatic mode easily isolated mononuclear cell from tissue of gentle, safety, timesaving, standard
Suspension, and tissue block is prepared into into homogenate, U.S. company BD is proposed Medimachine tissue samples preparing instruments, and Germany is beautiful
Its Ni company is proposed gentle MACS tissue separators.
Gentle MACS tissue separators use special disposable separating pipe.Particular design is carried in the lid of the pipe
Rotor and stator, there is the aperture of diaphragm seals at center.Mainly there are two kinds of separating pipes, C pipes and M pipes.C pipes rotor and stator it is only
Special geometry designs allow it effectively, leniently to process institutional framework, obtain the living cells of high-recovery.M pipes can be with treatment group
Block and cell are knitted, for separating subcellular components, such as nucleic acid.There is the aperture of a diaphragm seals at the center of both pipes,
Can guarantee that aseptically, sample is safely processed in closed system.Completely without opening test tube cap, you can addition enzyme
And other solution, either take out cell or the homogenate solution separated.
BD companies release product Medimachine tissue samples preparing instruments be will be cut into using the method for mechanical shearing it is little
The solid tissue of block is prepared into the automated system of unicellular or slender karyon suspension.Sample Preparation Procedure is not required to plus any digestion
Enzyme or detergent etc., it is made up of the part of Medimachine, Medicon and Filcon 3.Wherein Medimachine is main frame,
Afterwards both are accessory.Medicon is a round polyethylene element, and bottom is provided with a fixed stainless steel mesh screen, above there is about 100
Individual regular hexagon sieve aperture（Diameter is about 1 mm）, there have 6 specially designed microknifes around each sieve aperture to be fine to carry out
Tissue cutting, main frame then can drive the revolving fragment of Medicon axis to rotate with 100 turns per minute of rotating speed.Filcon is one
The cell filter of the cm of diameter 1.5, can be filtered by different pore size to histocyte suspension.
Said method is both needed to that tissue block is cut into into 1.0mm with manpower before cell separation3The tissue fragments of size, both
Arduously, it is time-consuming and not uniform enough, also add the probability of pollution.
The content of the invention
The present invention seeks to：A kind of histocyte automatic segregator simple and efficient to handle is provided.
The technical scheme is that：
A kind of histocyte automatic segregator, histocyte is separated into it is unicellular, the separator include separating pipe, institute
Separating pipe is stated including tube wall and the tube core in tube wall, lid is stamped in the tube wall top seal, bottom, and the tube core includes
Stator sleeve pipe and the rotor in sleeve pipe, the lid is provided with rotor connection, and rotor is connected with rotor connection, the separation
Device also includes being located at the motor outside separating pipe, and the motor provides rotary power by rotor connection for rotor.
Preferably, it is divided into the rotation of Archimedian screw area under control, cell separation area and negative pressure in the tube core from top to bottom
Area is cut, the Archimedian screw area under control collectively constitutes Archimedian screw pipe, cell by the screw rod of stator sleeve pipe and rotor
There is intensive tiny tusk Disengagement zone built with separation bar in separation bar, negative pressure rotary-cut area is built with rotary-cut cutter head, the spiral shell
Swing arm, separation bar and rotary-cut cutter head are sequentially connected composition rotor, rotary-cut cutter head bottom connection rotor connection.
Preferably, the stator casing part correspondence peeling knife head region is provided with some equally distributed holes.
It is further preferred that a diameter of 0.8-1.2mm of described hole.
Preferably, the rotor is sealed by rotor connection with the joint of lid.
Preferably, it is additionally provided with protection stay-warm case outside the separating pipe.
It is further preferred that the protection stay-warm case is built with temp probe and thermoelectric module, the temp probe and
Thermoelectric module is connected respectively with a microprocessor.
It is further preferred that being also equipped with a circulating fan being connected with microprocessor in the protection stay-warm case.
Preferably, the microprocessor is also connected to motor.
It is further preferred that being also connected with a touch screen on the microprocessor.
It is an advantage of the invention that：
1. histocyte automatic segregator provided by the present invention, using the automatic system of special tube core, microprocessor is fixed
The special application program of justice（Time, speed, direction）Operational procedure is optimized, sample can be easily processed, cell separation is made
Preparation process standardization, sequencing, reduce the impact of human factor, and change operator's dependence is into non-operator dependence, and institute
Tissue samples are needed only need to be simply prepared into the tissue block of several millimeters size before separation, separator can be peeled into automatically
1.0mm3The tissue fragments of size, and then isolated single cell suspension, for cell sorting, analysis or culture.Biopsy
And microscopy tissue is applicable.
2. the present invention has protection stay-warm case, temperature automatically controlled, plays beneficial to digestive enzyme activity, while ensureing user security
Property, realize sterile working.
3. the present invention is cut using rotary-cutting type sample, uniform in size.
4. present invention operation is gentle and simple and efficient.
Description of the drawings
Below in conjunction with the accompanying drawings and embodiment the invention will be further described：
Fig. 1 is the structural principle of histocyte separator of the present invention；
Fig. 2 is the structural representation of the separating pipe of the present invention；
Fig. 3 is the separating pipe structure decomposition figure shown in Fig. 2；
Fig. 4 is the schematic diagram of the temp probe of the present invention.
Specific embodiment
Embodiment：
As shown in figure 1, disclosed tissue to be separated into single celled histocyte automatic segregator, including point
From pipe, separating pipe lower section is equipped with motor, and motor speed is adjustable, can invert, can be according to set program（Time, speed, side
To）Operated.Protection stay-warm case is additionally provided with outside the separating pipe, the protection stay-warm case is built with temp probe, thermoelectricity
Cooling piece and circulating fan, the temp probe, thermoelectric module and circulating fan are connected respectively with a microprocessor, realize anti-
Temperature automatically controlled 37 DEG C in shield stay-warm case, play beneficial to digestive enzyme activity.Microprocessor is also connected with motor, the rotation of controlled motor,
A touch screen is also connected with the microprocessor, for being input into operational factor and showing situation in running.
As shown in Figures 2 and 3, the separating pipe includes tube wall 1 and the tube core 2 in tube wall, the tube wall top seal,
Lid 3 is stamped in bottom, and the tube core 2 includes stator sleeve pipe 4 and the rotor 5 in sleeve pipe 4, and the lid 3 is provided with rotor
Joint, rotor is connected with rotor connection, and the rotor is sealed by rotor connection with the joint of lid, the electricity below separating pipe
Machine provides rotary power by rotor connection for rotor, and stator sleeve pipe and tube wall are fixed on motor by bayonet socket.
The tube core 2 is divided into from top to bottom Archimedian screw area under control, cell separation area and negative pressure rotary-cut area, tube core
Stator casing part correspondence peeling knife head region be provided with the hole 6 of some equally distributed a diameter of 0.8-1.2mm.Ah
Built with screw rod 51, built with separation bar 52, negative pressure rotary-cut area is built with peeling knife in cell separation area in base Mead spiral area under control
53, the screw rod 51, separation bar 52 and rotary-cut cutter head 53 are sequentially connected composition rotor 5, and the connection of the bottom of rotary-cut cutter head 53 turns
Sub- joint.Wherein：
Archimedian screw area under control：The screw-rod structure of rotor collectively constitutes Archimedian screw pipe with stator sleeve pipe, utilizes
Sleeve pipe cylinder inside spin rotation rises and lifts liquid, and its operation principle is that, when inside spin is rotated, screw rod is on the one hand around itself
Axis rotation, on the other hand it roll along sleeve inner surface again, then form the sealed chamber of liquid.Screw rod each rotation,
Liquid in annular seal space pushes ahead a pitch, and with the continuous rotation for writing screw rod, liquid presses to another from an annular seal space
Annular seal space, finally extrudes liquid from cannula tip, meanwhile, negative pressure is formed in sleeve bottom, cause suction.
Cell separation area：Tissue fragments rise, through cell separation area, quilt under the extracting effect of negative pressure with liquid stream
Rotor middle part separation bar on intensive tiny dentalation it is soft be separated into individual cells.
Negative pressure rotary-cut area：The suction that negative pressure is formed makes tissue block be close to casing wall, and sinks into hole, at a high speed the rotation of rotation
Cutting knife head gets off the tissue cutting being absorbed in from hole, forms 1.0mm3The tissue fragments of size.
Liquid in separating pipe carries tissue fragments and constantly enters from the aperture of sleeve bottom, respectively through negative pressure rotary-cut
Area, cell separation area and Archimedian screw area under control, flow out from cannula tip, so go round and begin again, most tissue block dispersion at last
For individual cells.Umklapp process is designed with histocyte separable programming, when rotor is inverted, the liquid in tube core flows to inverse
Turn, the hole in tissue pieces blocking negative pressure rotary-cut area can be avoided.
The detached sample quality scope of the present invention：20-4000mg；Rotor speed：50-1000rpm, can both forward and reverse directions rotation
Turn；Time：The 1-1200 seconds；Temperature：Temperature controllable is in 37 DEG C；Application program：Carried according to different histocyte separation requirements
For the operational procedure and separable programming of optimization；Sample disposal：It is closed space in separating pipe.
The thermoelectric module of the present invention, is a kind of heat pump, and the work operating of semiconductor chilling plate is to use DC current, and it was both
Can freeze and can heat, determine to realize refrigeration on same cooling piece by changing the polarity of DC current or heat, this
The generation of effect is exactly the principle by thermoelectricity, when the galvanic couple that direct current is connected into by two kinds of different semi-conducting materials,
The two ends of galvanic couple can respectively absorb heat and release heat, can realize the purpose freezed or heat.
The temp probe temperature measuring head of the present invention adopts digital temperature sensor.This sensor becomes temperature sensing, signal
Change, A/D conversion etc. function be integrated on a chip, its internal structure is as shown in Figure 4.The chip is by temperature transition into telecommunications
Number, the electric signal initially enters small signal amplifier and is amplified；Subsequently into the A/D converter of 14；Jing after most
Cross two-wire serial digital interface output digit signals.This sensor has a calibration register to calibrate system in measurement process
Number can automatically calibrate the signal from sensor.
The present invention is simple and efficient to handle, makes sample preparation procedure standardization, sequencing, reduces the impact of human factor, becomes
Operator's dependence is non-operator dependence, and required tissue samples only need to simply be prepared into before separation several millimeters size
Tissue block, separator can be automatically peeled into 1.0mm3The tissue fragments of size, so it is isolated unicellular outstanding
Liquid, biopsy and microscopy are organized applicable.Prepared cell is applicable not only to FCM, can also carry out histochemical analysis and cell training
Support.
Above-described embodiment technology design only to illustrate the invention and feature, its object is to allow person skilled in the art
Will appreciate that present disclosure and implement according to this, can not be limited the scope of the invention with this.It is all main according to the present invention
The modification for wanting the Spirit Essence of technical scheme to be done, all should be included within the scope of the present invention.
Claims (8)
1. a kind of histocyte automatic segregator, tissue is separated into unicellular, and the separator includes separating pipe, and its feature exists
In：The separating pipe includes tube wall and the tube core in tube wall, and lid, the pipe are stamped in the tube wall top seal, bottom
Core includes stator sleeve pipe and the rotor in sleeve pipe, and the lid is provided with rotor connection, and rotor is connected with rotor connection, institute
Stating separator also includes being located at the motor outside separating pipe, and the motor provides rotary power by rotor connection for rotor；
Be divided into Archimedian screw area under control, cell separation area and negative pressure rotary-cut area in the tube core from top to bottom, Ah
Built with screw rod, built with separation bar, negative pressure rotary-cut area is built with rotary-cut cutter head, institute in cell separation area in base Mead spiral area under control
State screw rod, separation bar and rotary-cut cutter head and be sequentially connected composition rotor, rotary-cut cutter head bottom connection rotor connection；
The stator casing part correspondence peeling knife head region is provided with some equally distributed holes.
2. histocyte automatic segregator according to claim 1, it is characterised in that：A diameter of 0.8- of described hole
1.2mm。
3. histocyte automatic segregator according to claim 1, it is characterised in that：The rotor by rotor connection with
The joint sealing of lid.
4. the histocyte automatic segregator according to claim 1-3 any one, it is characterised in that：Outside the separating pipe
Portion is additionally provided with protection stay-warm case.
5. histocyte automatic segregator according to claim 4, it is characterised in that：The protection stay-warm case is built with temperature
Degree probe and thermoelectric module, the temp probe and thermoelectric module are connected respectively with a microprocessor.
6. histocyte automatic segregator according to claim 5, it is characterised in that：It is also equipped with the protection stay-warm case
One circulating fan being connected with microprocessor.
7. the histocyte automatic segregator according to claim 5 or 6, it is characterised in that：The microprocessor is also connected with
To motor.
8. histocyte automatic segregator according to claim 7, it is characterised in that：One is also connected with the microprocessor
Touch screen.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410772430.5A CN104568539B (en) | 2014-12-16 | 2014-12-16 | Automatic histocyte separator |
| PCT/CN2015/077135 WO2016095390A1 (en) | 2014-12-16 | 2015-04-22 | Automatic histocyte separator |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410772430.5A CN104568539B (en) | 2014-12-16 | 2014-12-16 | Automatic histocyte separator |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104568539A CN104568539A (en) | 2015-04-29 |
| CN104568539B true CN104568539B (en) | 2017-05-10 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410772430.5A Active CN104568539B (en) | 2014-12-16 | 2014-12-16 | Automatic histocyte separator |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN104568539B (en) |
| WO (1) | WO2016095390A1 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018088959A1 (en) * | 2016-11-10 | 2018-05-17 | Celligenics Pte. Ltd | Device for disrupting tissue |
| JP7064191B2 (en) * | 2018-02-21 | 2022-05-10 | エイブル株式会社 | Cell dispersal method and cell disperser |
| WO2021208012A1 (en) * | 2020-04-16 | 2021-10-21 | Suzhou Singleron Biotechnologies, Ltd. | Tissue dissociation tube and instrument |
| CN112067408A (en) * | 2020-10-16 | 2020-12-11 | 昆明东环科技有限公司 | Automatic cell block preparation machine and method |
| CN113174361B (en) * | 2021-06-10 | 2023-11-14 | 南通市第一老年病医院(上海大学附属南通医院、南通市第六人民医院、南通市肺科医院) | Pathological cell separation adsorption type extraction method |
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|---|---|---|---|---|
| CN1580732A (en) * | 2003-08-07 | 2005-02-16 | 中国科学院海洋研究所 | Shellfish sample preparing method suitable for detecting ploidy by flow cytometry |
| CN101957282A (en) * | 2010-10-20 | 2011-01-26 | 杭州市中医院 | Preparation method and application of single cell or tissue particle suspension |
| CN101983328A (en) * | 2008-03-31 | 2011-03-02 | 希森美康株式会社 | Cell processing device, sample preparing device, and cell analyzing device |
| US8871159B1 (en) * | 2013-12-17 | 2014-10-28 | Christian Apfel | Preparation of cells, cell aggregates and tissue fragments |
| CN204286897U (en) * | 2014-12-16 | 2015-04-22 | 何向锋 | A kind of histocyte automatic segregator |
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|---|---|---|---|---|
| TWI255738B (en) * | 2003-12-31 | 2006-06-01 | Ind Tech Res Inst | Tissue pulverizer |
| US8491497B2 (en) * | 2010-05-13 | 2013-07-23 | Ethicon Endo-Surgery, Inc. | Method and apparatus for morcellating tissue |
| CN102220224B (en) * | 2011-05-19 | 2014-10-01 | 汪华 | Automatic medical cell separation device |
| JP5848541B2 (en) * | 2011-07-26 | 2016-01-27 | 日本光電工業株式会社 | Cell isolation instrument |
| CN203639459U (en) * | 2013-09-12 | 2014-06-11 | 博雅干细胞科技有限公司 | Tissue processor for separating living cells |
| CN104498350B (en) * | 2014-12-16 | 2016-07-06 | 何向锋 | A kind of homogenizer for separating subcellular components |
-
2014
- 2014-12-16 CN CN201410772430.5A patent/CN104568539B/en active Active
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2015
- 2015-04-22 WO PCT/CN2015/077135 patent/WO2016095390A1/en active Application Filing
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1580732A (en) * | 2003-08-07 | 2005-02-16 | 中国科学院海洋研究所 | Shellfish sample preparing method suitable for detecting ploidy by flow cytometry |
| CN101983328A (en) * | 2008-03-31 | 2011-03-02 | 希森美康株式会社 | Cell processing device, sample preparing device, and cell analyzing device |
| CN101957282A (en) * | 2010-10-20 | 2011-01-26 | 杭州市中医院 | Preparation method and application of single cell or tissue particle suspension |
| US8871159B1 (en) * | 2013-12-17 | 2014-10-28 | Christian Apfel | Preparation of cells, cell aggregates and tissue fragments |
| CN204286897U (en) * | 2014-12-16 | 2015-04-22 | 何向锋 | A kind of histocyte automatic segregator |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2016095390A1 (en) | 2016-06-23 |
| CN104568539A (en) | 2015-04-29 |
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Effective date of registration: 20170510 Address after: Tongzhou District Pingchao town in Jiangsu province 226000 Nantong Tongyang Road No. 30 Co-patentee after: He Xiangfeng Patentee after: Nantong Tumor Hospital Address before: Tongzhou District Pingchao town in Jiangsu province 226000 Nantong Tongyang Road No. 30 Co-patentee before: Nantong Tumor Hospital Patentee before: He Xiangfeng |
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Effective date of registration: 20190722 Address after: 226010 A1 Floor, Jianghai Zhihui Park, 266 New Century Avenue, Nantong High-tech Zone, Jiangsu Province Patentee after: JIANGSU XINGRUI BIOLOGICAL TECHNOLOGY Co.,Ltd. Address before: Tongzhou District Pingchao town in Jiangsu province 226000 Nantong Tongyang Road No. 30 Co-patentee before: He Xiangfeng Patentee before: Nantong Tumor Hospital |
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Effective date of registration: 20221208 Address after: 259 beigaoxin Road, Tangzha Town, Chongchuan District, Nantong City, Jiangsu Province, 226000 Patentee after: Jiangsu sailana biomedical Co.,Ltd. Address before: 226010 A1 / F, Jianghai Zhihui garden, 266 New Century Avenue, Nantong high tech Zone, Jiangsu Province Patentee before: JIANGSU XINGRUI BIOLOGICAL TECHNOLOGY Co.,Ltd. |