CN104568539A - Automatic histocyte separator - Google Patents
Automatic histocyte separator Download PDFInfo
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- CN104568539A CN104568539A CN201410772430.5A CN201410772430A CN104568539A CN 104568539 A CN104568539 A CN 104568539A CN 201410772430 A CN201410772430 A CN 201410772430A CN 104568539 A CN104568539 A CN 104568539A
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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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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, particularly a kind of histocyte automatic segregator.
Background technology
Histocyte is the important materials of carrying out the life sciences such as cell biology, molecular biology, immunology, science of heredity, oncology, such as carry out flow cytometry (flow cytometry, FCM) namely the prerequisite detected requires that sample is single cell suspension, and most of solid tissue is all difficult to make high-quality single cell suspension sample, thus greatly limit FCM to the solid tissue particularly analysis of solid tumor and research.Histocyte separation vessel is used for body tissue as lung, liver, tumour etc. are separated into individual cells suspension, tradition solid tissue being separated into individual cells is prepared strategy and is generally adopted manual method, mainly contains mechanical shearing method, enzyme digestion, detergent method of chemical treatment, syringe needle suction method etc.
In order to gentleness, safety, timesaving, standard automatic mode easily from tissue isolated mononuclear cell suspension, and tissue block is prepared into homogenate, U.S. company BD is proposed Medimachine tissue samples preparing instrument, and German Mei Tian Ni company is proposed gentle MACS and organizes separation vessel.
Gentle MACS organizes separation vessel to use special disposable separator tube.The pipe of this pipe covers rotor with particular design and stator, and there is the aperture of diaphragm seals at center.Mainly contain two kinds of separator tubes, C pipe and M pipe.Unique geometry designs of C pipe rotor and stator makes it effectively, leniently can process institutional framework, obtains the living cells of high-recovery.M pipe can process tissue block and cell, for separating of subcellular components, and such as nucleic acid.There is the aperture of a diaphragm seals at the center of these two kinds of pipes, can ensure aseptically, processing sample safely in closed system.Do not need completely to open test tube cap, enzyme and other solution can be added, or take out the cell or homogenate solution separated.
The product Medimachine tissue samples preparing instrument that BD company releases is the automated system adopting the method for mechanical shearing the solid tissue be cut into small pieces to be prepared into unicellular or slender karyon suspension.Sample Preparation Procedure does not need to add any digestive ferment or detergent etc., it by Medimachine, Medicon and Filcon 3 part form.Wherein Medimachine is main frame, is accessory both rear.Medicon is a round polyethylene element, bottom is provided with a fixing stainless steel mesh screen, on to have an appointment 100 regular hexagon sieve apertures (diameter is about 1 mm), have 6 around each sieve aperture through the microknife of particular design to carry out the cutting of meticulous tissue, main frame then can the rotating speed of 100 turns per minute drive the revolving fragment of Medicon axis to rotate.Filcon is the cell filter of diameter 1.5 cm, can filter by different pore size to histocyte suspension.
Tissue block all need be cut into 1.0mm with manpower by said method before cell separation
3the tissue fragments of size, both required great effort, time-consuming, even not again, also add the probability of pollution.
Summary of the invention
The present invention seeks to: a kind of histocyte automatic segregator simple and efficient to handle is provided.
Technical scheme of the present invention is:
A kind of histocyte automatic segregator, histocyte is separated into unicellular, described separation vessel comprises separator tube, and described separator tube comprises tube wall and is positioned at the tube core of tube wall, described tube wall top seal, pipe lid is stamped in bottom, described tube core comprises stator sleeve pipe and is arranged in the rotor of sleeve pipe, and described pipe covers and is provided with rotor connection, and rotor is connected with rotor connection, described separation vessel also comprises the motor being positioned at separator tube outside, described motor by rotor connection for rotor provides rotary power.
Preferably, Archimedean screw area under control, cell separation district and negative pressure rotary-cut district is divided into from top to bottom successively in described tube core, described Archimedean screw area under control forms Archimedean screw pipe jointly by the screw rod of stator sleeve pipe and rotor, cell separation district is built with separation bar, separation bar there is intensive tiny tusk, negative pressure rotary-cut district is built with rotary-cut cutter head, and described screw rod, separation bar and rotary-cut cutter head connect to form rotor successively, connect rotor connection bottom rotary-cut cutter head.
Preferably, the corresponding peeling knife head region of described stator casing part is provided with some equally distributed holes.
Preferred further, the diameter of described hole is 0.8-1.2mm.
Preferably, described rotor is sealed by the joint of rotor connection and pipe lid.
Preferably, described separator tube outside is also provided with protection stay-warm case.
Preferred further, described protection stay-warm case is built with temp probe and thermoelectric module, and described temp probe and thermoelectric module are connected with a microprocessor respectively.
Preferred further, a circulating fan be connected with microprocessor is also housed in described protection stay-warm case.
Preferably, described microprocessor is also connected to motor.
Preferred further, described microprocessor also connects a touch screen.
Advantage of the present invention is:
1. histocyte automatic segregator provided by the present invention, use the automatic system of special tube core, microprocessor defines special application program (time, speed, direction) by working specification optimization, sample can be processed easily, make the standardization of cell separation preparation process, sequencing, reduce the impact of human factor, becoming operator's dependence is not operation person dependence, and required tissue samples only simply need be prepared into the tissue block of several mm in size before separation, separation vessel can be peeled into 1.0mm automatically
3the tissue fragments of size, and then separation obtains single cell suspension, for cell sorting, analysis or cultivation.Biopsy and microscopy are organized all applicable.
2. the present invention has protection stay-warm case, temperature automatically controlled, is beneficial to digestive enzyme activity and plays, ensure user security simultaneously, realize sterile working.
3. the present invention adopts rotary-cutting type sample to cut, and size is even.
4. the present invention operates gentleness, and simple and efficient.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the invention will be further described:
Fig. 1 is the structural principle of histocyte separation vessel of the present invention;
Fig. 2 is the structural representation of separator tube of the present invention;
Fig. 3 is the separator tube STRUCTURE DECOMPOSITION figure shown in Fig. 2;
Fig. 4 is the schematic diagram of temp probe of the present invention.
Embodiment
embodiment:
As shown in Figure 1, disclosed is separated into single celled histocyte automatic segregator by tissue, comprise separator tube, below separator tube, motor is housed, motor speed is adjustable, can reverse, and can operate according to set program (time, speed, direction).Described separator tube outside is also provided with protection stay-warm case, described protection stay-warm case is built with temp probe, thermoelectric module and circulating fan, described temp probe, thermoelectric module and circulating fan are connected with a microprocessor respectively, realize protection stay-warm case in temperature automatically controlled 37 DEG C, be beneficial to digestive enzyme activity play.Microprocessor also connects motor, controls the rotation of motor, described microprocessor also connects a touch screen, is used for inputting situation in operational factor and display operational process.
As shown in Figures 2 and 3, described separator tube comprises tube wall 1 and is positioned at the tube core 2 of tube wall, described tube wall top seal, pipe lid 3 is stamped in bottom, and described tube core 2 comprises stator sleeve pipe 4 and is arranged in the rotor 5 of sleeve pipe 4, and described pipe lid 3 is provided with rotor connection, rotor is connected with rotor connection, described rotor is sealed by the joint of rotor connection and pipe lid, and the motor below separator tube is by rotor connection for rotor provides rotary power, and stator sleeve pipe and tube wall are fixed on motor by bayonet socket.
Described tube core 2 is divided into Archimedean screw area under control, cell separation district and negative pressure rotary-cut district from top to bottom successively, and the corresponding peeling knife head region of stator casing part of tube core is provided with the hole 6 that some equally distributed diameters are 0.8-1.2mm.Described Archimedean screw area under control is built with screw rod 51, cell separation district is built with separation bar 52, negative pressure rotary-cut district is built with rotary-cut cutter head 53, and described screw rod 51, separation bar 52 and rotary-cut cutter head 53 connect to form rotor 5 successively, connect rotor connection bottom rotary-cut cutter head 53.Wherein:
Archimedean screw area under control: the screw-rod structure of rotor forms Archimedean screw pipe jointly with stator sleeve pipe, utilize sleeve pipe cylinder inside spin to take turns turn rising and promote liquid, its principle of work is when inside spin rotates, screw rod rotates around the axis of itself on the one hand, it rolls along sleeve inner surface again on the other hand, so form the sealed chamber of liquid.Screw rod often circles, and the liquid in annular seal space pushes ahead a pitch, and with the continuous rotation of work screw rod, liquid presses to another annular seal space from an annular seal space, is finally extruded from cannula tip by liquid, meanwhile, forms negative pressure, cause suction at sleeve bottom.
Cell separation district: tissue fragments under the extracting effect of negative pressure, with liquid stream rise, through cell separation district, by the intensive tiny dentalation in separation bar in the middle part of rotor soft be separated into individual cells.
Negative pressure rotary-cut district: the suction that negative pressure is formed makes tissue block be close to casing wall, and sinks into hole, and the tissue cutting be absorbed in from hole gets off by the rotary-cut cutter head of High Rotation Speed, forms 1.0mm
3the tissue fragments of size.
Liquid in separator tube carries tissue fragments and constantly enters from the aperture of sleeve bottom, and respectively through negative pressure rotary-cut district, cell separation district and Archimedean screw area under control, flow out from cannula tip, so go round and begin again, tissue block is separated into individual cells the most at last.Be designed with Umklapp process in histocyte separable programming, when rotor reverses, the flow direction in tube core reverses, and tissue pieces can be avoided to block the hole in negative pressure rotary-cut district.
The sample quality scope that the present invention is separated: 20-4000mg; Rotor speed: 50-1000rpm, can rotate by both forward and reverse directions; Time: 1-1200 second; Temperature: temperature controllable is in 37 DEG C; Application program: working specification and separable programming that optimization is provided according to different histocyte separation requirement; Sample disposal: be airtight space in separator tube.
Thermoelectric module of the present invention, it is a kind of heat pump, the work running of semiconductor chilling plate is by DC current, it not only can freeze but also can heat, decide on same cooling piece, realize refrigeration or heating by the polarity changing DC current, the generation of this effect is exactly the principle by thermoelectricity, when the galvanic couple that direct current is connected into by two kinds of different semiconductor materials, heat can be absorbed at the two ends of galvanic couple respectively and release heat, the object of freezing or heating can be realized.
Temp probe temperature measuring head of the present invention adopts digital temperature sensor.This sensor is by function i ntegration such as temperature sensing, signal conversion, A/D conversions on a chip, and its inner structure as shown in Figure 4.Temperature transition is become electric signal by this chip, and first this electric signal enters small signal amplifier and amplify; Then the A/D converter of 14 is entered; Eventually pass two-wire serial digital interface output digit signals.This sensor has a calibration register can in measuring process, and calibration factor can carry out the signal of sensor by automatic calibration.
The present invention is simple and efficient to handle, make sample preparation procedure standardization, sequencing, reduce the impact of human factor, becoming operator's dependence is not operation person dependence, and required tissue samples only simply need be prepared into the tissue block of several mm in size before separation, separation vessel can be peeled into 1.0mm automatically
3the tissue fragments of size, and then separation obtains single cell suspension, biopsy and microscopy are organized all applicable.Prepared cell is not only applicable to FCM, also can carry out histochemical analysis and cell chulture etc.
Above-described embodiment, only for technical conceive of the present invention and feature are described, its object is to person skilled in the art can be understood content of the present invention and implement according to this, can not limit the scope of the invention with this.All modifications done according to the Spirit Essence of main technical schemes of the present invention, all should be encompassed within protection scope of the present invention.
Claims (10)
1. a histocyte automatic segregator, tissue is separated into unicellular, described separation vessel comprises separator tube, it is characterized in that: described separator tube comprises tube wall and is positioned at the tube core of tube wall, described tube wall top seal, pipe lid is stamped in bottom, described tube core comprises stator sleeve pipe and is arranged in the rotor of sleeve pipe, described pipe covers and is provided with rotor connection, rotor is connected with rotor connection, described separation vessel also comprises the motor being positioned at separator tube outside, described motor by rotor connection for rotor provides rotary power.
2. histocyte automatic segregator according to claim 1, it is characterized in that: in described tube core, be divided into Archimedean screw area under control, cell separation district and negative pressure rotary-cut district from top to bottom successively, described Archimedean screw area under control is built with screw rod, cell separation district is built with separation bar, negative pressure rotary-cut district is built with rotary-cut cutter head, described screw rod, separation bar and rotary-cut cutter head connect to form rotor successively, connect rotor connection bottom rotary-cut cutter head.
3. histocyte automatic segregator according to claim 2, is characterized in that: the corresponding peeling knife head region of described stator casing part is provided with some equally distributed holes.
4. histocyte automatic segregator according to claim 3, is characterized in that: the diameter of described hole is 0.8-1.2mm.
5. histocyte automatic segregator according to claim 1, is characterized in that: described rotor is sealed by the joint of rotor connection and pipe lid.
6. the histocyte automatic segregator according to claim 1-5 any one, is characterized in that: described separator tube outside is also provided with protection stay-warm case.
7. histocyte automatic segregator according to claim 6, is characterized in that: described protection stay-warm case is built with temp probe and thermoelectric module, and described temp probe and thermoelectric module are connected with a microprocessor respectively.
8. histocyte automatic segregator according to claim 7, is characterized in that: a circulating fan be connected with microprocessor is also housed in described protection stay-warm case.
9. the histocyte automatic segregator according to claim 7 or 8, is characterized in that: described microprocessor is also connected to motor.
10. histocyte automatic segregator according to claim 9, is characterized in that: described microprocessor also connects a 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 true CN104568539A (en) | 2015-04-29 |
| CN104568539B CN104568539B (en) | 2017-05-10 |
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| 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) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110088590A (en) * | 2016-11-10 | 2019-08-02 | 赛利詹尼克斯私人投资有限公司 | For rupturing the device of tissue |
| JP2019140985A (en) * | 2018-02-21 | 2019-08-29 | エイブル株式会社 | Cell dispersion method and cell dispersion apparatus |
| CN113174361A (en) * | 2021-06-10 | 2021-07-27 | 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 |
| WO2021208012A1 (en) * | 2020-04-16 | 2021-10-21 | Suzhou Singleron Biotechnologies, Ltd. | Tissue dissociation tube and instrument |
| WO2022078235A1 (en) * | 2020-10-16 | 2022-04-21 | 赵稳兴 | Automatic cell block preparation machine and 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 |
| 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 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110088590A (en) * | 2016-11-10 | 2019-08-02 | 赛利詹尼克斯私人投资有限公司 | For rupturing the device of tissue |
| CN110088590B (en) * | 2016-11-10 | 2022-04-26 | 赛利詹尼克斯私人投资有限公司 | Device for rupturing tissue |
| JP2019140985A (en) * | 2018-02-21 | 2019-08-29 | エイブル株式会社 | Cell dispersion method and cell dispersion apparatus |
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| WO2021208012A1 (en) * | 2020-04-16 | 2021-10-21 | Suzhou Singleron Biotechnologies, Ltd. | Tissue dissociation tube and instrument |
| WO2022078235A1 (en) * | 2020-10-16 | 2022-04-21 | 赵稳兴 | Automatic cell block preparation machine and method |
| CN113174361A (en) * | 2021-06-10 | 2021-07-27 | 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 |
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104568539B (en) | 2017-05-10 |
| WO2016095390A1 (en) | 2016-06-23 |
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