CN100562739C - Target automatic separating apparatus based on immunomagnetic beads - Google Patents

Target automatic separating apparatus based on immunomagnetic beads Download PDF

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Publication number
CN100562739C
CN100562739C CNB2007100707101A CN200710070710A CN100562739C CN 100562739 C CN100562739 C CN 100562739C CN B2007100707101 A CNB2007100707101 A CN B2007100707101A CN 200710070710 A CN200710070710 A CN 200710070710A CN 100562739 C CN100562739 C CN 100562739C
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China
Prior art keywords
sample
chip microcomputer
immunomagnetic beads
mouth
target
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Expired - Fee Related
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CNB2007100707101A
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Chinese (zh)
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CN101118202A (en
Inventor
叶尊忠
应义斌
王剑平
李延斌
饶秀勤
蒋焕煜
蒋雪松
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Zhejiang University ZJU
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Zhejiang University ZJU
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Abstract

The invention discloses a kind of target automatic separating apparatus based on immunomagnetic beads.Comprise mixed cell, separative element and control module.Detected sample and immunomagnetic beads are joined in the sample hose, sample hose is under the various combination of at the uniform velocity rotation, speed change rotation, static enrichment or forward and reverse rotation status, and antibody fixing on target in the detected sample in the sample hose and the immunomagnetic beads produces specific reaction; Under fixed magnetic field, immunomagnetic beads be adsorbed on the sample hose inwall by the target in the immune magnetic detected sample captured; Collect immunomagnetic beads and, be used for biological detection or shop dish by the target in the immune magnetic detected sample captured.The present invention produces specific reaction by antibody fixing on the detected target in the sample hose and the immunomagnetic beads can accurately separate and screen the target detection thing, and the combination by the different blended syntype reaches optimum mixed effect, is widely used in the pre-treatment of sample.

Description

Target automatic separating apparatus based on immunomagnetic beads
Technical field
The present invention relates to a kind of target automatic separating apparatus based on immunomagnetic beads.
Background technology
As everyone knows, sample pre-treatments is very important means, and its success or not is most important to the detection of sample.Utilize immune magnetic particle (magnetic bead) to screen and be separated in food, clinical, soil obtains more and more certainly with the method for targets such as the microorganism in the environment, cell, chemical element.Owing to quick, the highly enriched ratio of immunomagnetic beads separation/screening technique, to advantages such as equipment requirements are simple, use very wide in practice.At present, immunomagnetic beads separation/screening technique generally is made up of two steps: mix and separate.General mixer can only be by the rotation of direction, and rotational speed is manual adjustments, and incorporation time can only be by artificial control; Separation equipment generally is made up of permanent magnet and support, and disengaging time also is by manually controlling.Because these two committed steps are independent operations, make separating effect be subjected to the influence of all many-sides, and must buy a plurality of equipment and just can finish the first separation operation, these factors make the pre-treatment effect be subjected to very big influence.
Summary of the invention
The invention provides and a kind ofly be used for separating automatically the device of detected target based on immunomagnetic beads.
The technical solution used in the present invention is as follows:
One, based on the target separation method of immunomagnetic beads, the step of this method is as follows: detected sample and immunomagnetic beads are joined in the sample hose, sample hose is under the various combination of at the uniform velocity rotation, speed change rotation, static enrichment or forward and reverse rotation status, and antibody fixing on target in the detected sample in the sample hose and the immunomagnetic beads produces specific reaction; Under fixed magnetic field, immunomagnetic beads be adsorbed on the sample hose inwall, draw the damping fluid that cleans sample hose after the supernatant and add small size by the target in the immune magnetic detected sample captured; Collect immunomagnetic beads and, be used for biological detection or shop dish by the target in the immune magnetic detected sample captured.
Two, based on the target tripping device of immunomagnetic beads, comprise mixed cell, separative element and control module; Wherein:
1) mixed cell, n sample hose are separately fixed on the biased sample plate in separately the sample well, and hybrid electric motor is rotationally connected by first shaft joint and biased sample plate; Hybrid electric motor is electrically connected by umbilical cable and control module;
2) separative element, n piece discrete magnets are separately fixed at and separate on the driver plate, n piece discrete magnets be fixed on the mixed drive plate on n sample hose corresponding one by one, the separation motor by second shaft joint with separate driver plate and be rotationally connected; Separating motor is electrically connected by umbilical cable and control module;
3) control module, single-chip microcomputer link to each other with LCD, keyboard, buffering drive circuit, crystal oscillating circuit, parameter storage, communication interface, alarm unit respectively; Buffering drive circuit connects hybrid electric motor respectively and separates motor; Communication interface links to each other with computing machine through usb communication line, power circuit respectively with power supply input, single-chip microcomputer, separate motor and link to each other with hybrid electric motor.
The X1 of AT89C52 single-chip microcomputer, X2 mouth are connected with crystal oscillating circuit; The buffering drive circuit that the P0 mouth of single-chip microcomputer is formed via 74HC573 chip and ULN2803A chip is respectively with hybrid electric motor with to separate motor continuous; The P2 mouth of single-chip microcomputer links to each other with the GDM12864 LCD by extended chip 74HC595 with P3.4, P3.6, P3.7; The P1 mouth of single-chip microcomputer links to each other with 4 * 4 keyboards; The RXD of single-chip microcomputer, TXD mouth link to each other with communication interface; The P3.2 of single-chip microcomputer, P3.3 mouth link to each other with parameter storage; The P3.5 mouth of single-chip microcomputer links to each other with alarm unit.
The beneficial effect that the present invention has is: produce specific reaction by antibody fixing on the detected target in the sample hose and the immunomagnetic beads and can accurately separate and screen the target detection thing, and the combination by the different blended syntype reaches optimum mixed effect, is widely used in the pre-treatment of sample.
Description of drawings
Fig. 1 is the overall schematic of tripping device.
Fig. 2 is that the integral body of tripping device and control enclosure and sample mix plate revolve the synoptic diagram that turn 90 degrees.
Fig. 3 is that the integral body of tripping device is formed structural drawing.
Fig. 4 is the control circuit composition diagram of tripping device.
Fig. 5 is the control circuit schematic diagram of tripping device.
Among the figure: 1, mixed cell; 2, separative element; 3, control module; 101,102,103,104, sample hose; 105,106,107,108, sample well; 109, hybrid electric motor; 110, bearing; 111, biased sample plate; 112, shaft joint; 201,202,203,204, discrete magnets; 205, separate driver plate; 206, bearing; 207, shaft joint; 208, separate motor; 301,302, insulated support; 303, umbilical cable; 304, LCD; 305, keyboard; 306, control circuit board; 307, buffering drive circuit; 308, power supply input; 309, power circuit; 310, single-chip microcomputer; 311, crystal oscillating circuit; 312, parameter storage; 313, communication interface; 314, alarm unit; 315, usb communication line; 316, computing machine.
Embodiment
The invention will be further described below in conjunction with drawings and Examples.
As Fig. 1, Fig. 2, Fig. 3, shown in Figure 4, the present invention includes mixed cell 1, separative element 2 and control module 3; Wherein:
1) mixed cell 1,4 sample hoses 101,102,103,104, be separately fixed on the biased sample plate 111 in 105,106,107,108 sample wells separately, one end of biased sample plate 111 links to each other by first shaft joint 112 with hybrid electric motor 109, and the other end of biased sample plate 111 links to each other with shell by bearing 110; Hybrid electric motor 109 is electrically connected by umbilical cable 303 and control module 3;
2) separative element 2,4 discrete magnets are separately fixed at separates on the driver plate 205,4 discrete magnets be fixed on biased sample plate 111 on 4 sample hoses corresponding one by one, an end that separates driver plate 205 by shaft joint 207 with separate motor 208 and link to each other, the other end that separates driver plate 205 links to each other with shell by bearing 206; Separating motor 208 is electrically connected by umbilical cable 303 and control module 3;
3) control module 3, and single-chip microcomputer 310 links to each other with LCD 304, keyboard 305, buffering drive circuit 307, crystal oscillating circuit 311, parameter storage 312, communication interface 313, alarm unit 314 respectively; Buffering drive circuit 307 connects hybrid electric motor 109 respectively and separates motor 208; Communication interface 313 links to each other with computing machine 316 through usb communication line 315, power circuit 309 respectively with power supply input 308, single-chip microcomputer 310, separate motor 208 and link to each other with hybrid electric motor 109, control circuit board 306 usefulness 301 and 302 insulated supports rest on the housing.
As shown in Figure 5, the X1 of AT89C52 single-chip microcomputer 310, X2 mouth are connected with crystal oscillating circuit 311; The buffering drive circuit 307 that the P0 mouth of single-chip microcomputer 310 is formed via 74HC573 chip and ULN2803A chip is respectively with hybrid electric motor 109 with to separate motor 208 continuous; The P2 mouth of single-chip microcomputer 310 links to each other with GDM12864 LCD 304 by extended chip 74HC595 with P3.4, P3.6, P3.7; The P1 mouth of single-chip microcomputer 310 links to each other with 4 * 4 keyboards 305; The RXD of single-chip microcomputer 310, TXD mouth link to each other with communication interface 313; The P3.2 of single-chip microcomputer 310, P3.3 mouth link to each other with parameter storage 312; The P3.5 mouth of single-chip microcomputer 310 links to each other with alarm unit 314.LCD 304, it can show English character and Chinese character.
The course of work of the present invention is as follows:
(1) immunomagnetic beads and detected sample are joined in the sample hose 101,102,103,104, and close the cover;
(2) sample hose 101,102,103,104 is inserted in the sample well 105,106,107,108;
(3) import parameters such as incorporation time, disengaging time and mixed mode on keyboard 305, LCD 304 will show occurrence; After input finishes, the start button on the keypad 305;
(4) control circuit board 306 output signals control hybrid electric motor 109 drives biased sample plate 111 and drives sample hose 101,102,103,104 by the pattern rotation of setting; Rotatablely move and make immunomagnetic beads and the interior object of detected sample in the sample hose 101,102,103,104 fully mix;
(5) after blend step finishes, control circuit board 306 output signals make sample hose 101,102,103,104 be in initial upright position, the control of control circuit board 306 output signals separates motor 208, drive the fixing discrete magnets 201,202,203,204 onboard of separation driver plate 205 and drive and arrive the outer wall of sample hose 101,102,103,104, and stop operating; Under the action of a magnetic field of discrete magnets 201,202,203,204, separated and assembling near magnetic direction toward the inwall of sample hose 101,102,103,104 gradually by the object in the immunomagnetic beads institute detected sample captured, and begin agglomerating with solution;
(6) after step to be separated finished, the operator opened the lid of sample hose 101,102,103,104 and takes supernatant away with liquid-transfering gun; After taking supernatant away, the conclusion button on the keypad 305;
(7) control of control circuit board 306 output signals separates motor 208, drives to separate driver plate 205 and drive the outer wall that discrete magnets 201,202,203,204 is left sample hose 101,102,103,104; And return to initial position;
(8) take off in the sample hose 101,102,103,104, the damping fluid that adds small size just can obtain by immunomagnetic beads captured object thing solution.Can obtain the sample after the concentrating and separating like this, it can be used for subsequent detection such as biology sensor detection, ELISA, PCR, for they provide very effective sample and processing means, and has improved detectability.
Fig. 4 is the control circuit composition diagram of tripping device.In order to make computing machine 316 control magnetic separating devices, magnetic separating device is connected with computing machine 316 by usb communication line 315.The operation of computer software control magnetic separating device, and monitor the state of magnetic separating device in real time, make things convenient for user management.
Above-mentioned embodiment is used for the present invention that explains, rather than limits the invention, and in the protection domain of spirit of the present invention and claim, any modification and change to the present invention makes all fall into protection scope of the present invention.

Claims (2)

1, a kind of target automatic separating apparatus based on immunomagnetic beads is characterized in that comprising mixed cell (1), separative element (2) and control module (3); Wherein:
Mixed cell (1), n sample hose are separately fixed on the biased sample plate (111) in separately the sample well, and hybrid electric motor (109) is rotationally connected by first shaft joint (112) and biased sample plate (111); Hybrid electric motor (109) is electrically connected by umbilical cable (303) and control module (3);
Separative element (2), n piece discrete magnets is separately fixed at separates on the driver plate (205), n piece discrete magnets is corresponding one by one with n sample hose on being fixed on biased sample plate (111), separate motor (208) by second shaft joint (207) with separate driver plate (205) and be rotationally connected; Separating motor (208) is electrically connected by umbilical cable (303) and control module (3);
Control module (3), single-chip microcomputer (310) link to each other with LCD (304), keyboard (305), buffering drive circuit (307), crystal oscillating circuit (311), parameter storage (312), communication interface (313), alarm unit (314) respectively; Buffering drive circuit (307) connects hybrid electric motor (109) respectively and separates motor (208); Communication interface (313) links to each other with computing machine (316) through usb communication line (315), power circuit (309) respectively with power supply input (308), single-chip microcomputer (310), separate motor (208) and link to each other with hybrid electric motor (109).
2, a kind of target automatic separating apparatus based on immunomagnetic beads according to claim 1 is characterized in that: the X1 of single-chip microcomputer (310), X2 mouth are connected with crystal oscillating circuit (311); The buffering drive circuit (307) that the P0 mouth of single-chip microcomputer (310) is formed via 74HC573 chip and ULN2803A chip is respectively with hybrid electric motor (109) with to separate motor (208) continuous; The P2 mouth of single-chip microcomputer (310) links to each other with GDM12864 LCD (304) by extended chip 74HC595 with P3.4, P3.6, P3.7; The P1 mouth of single-chip microcomputer (310) links to each other with 4 * 4 keyboards (305); The RXD of single-chip microcomputer (310), TXD mouth link to each other with communication interface (313); The P3.2 of single-chip microcomputer (310), P3.3 mouth link to each other with parameter storage (312); The P3.5 mouth of single-chip microcomputer (310) links to each other with alarm unit (314).
CNB2007100707101A 2007-08-06 2007-08-06 Target automatic separating apparatus based on immunomagnetic beads Expired - Fee Related CN100562739C (en)

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* Cited by examiner, † Cited by third party
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CN101358979B (en) * 2008-08-18 2012-11-28 杭州师范大学 Chloramphenicol immune detecting system marked by magnetic bead
US10722879B2 (en) * 2013-11-25 2020-07-28 Gencell Biosystems Ltd. Magnetic separation
CN103861735B (en) * 2014-03-19 2017-01-25 东南大学 Magnetic separation device
CN103878061B (en) * 2014-03-21 2016-05-25 东南大学 A kind of full automatic magnetic separating device
CN104297039B (en) * 2014-09-25 2017-01-11 深圳市奥特库贝科技有限公司 Novel magnetic separation mechanism
CN104388377A (en) * 2014-11-24 2015-03-04 关节动力安达(天津)生物科技有限公司 Magnetic cell sorting method capable of regulating magnetic field
CN104655831A (en) * 2015-03-04 2015-05-27 哈尔滨工业大学(威海) Magnetic bead cleaning and separating device for full-automatic chemical luminescence immunity analyzer
CN104805010B (en) * 2015-04-22 2017-01-04 江苏微全芯生物科技有限公司 A kind of pathogenic bacterium fast enriching chip and preparation method thereof
CN105259355A (en) * 2015-09-23 2016-01-20 浙江大学 Automatic biological sample pretreatment device
CN106111329B (en) * 2016-08-16 2017-10-13 闫维新 A kind of actuation means of miniature magnetic bead
JP6782264B2 (en) * 2018-02-20 2020-11-11 株式会社日立ハイテク Automatic analyzer
CN108296013B (en) * 2018-03-02 2023-11-14 浙江大学 Separation device for superparamagnetic nanoparticles and application thereof
CN110229781A (en) * 2019-06-11 2019-09-13 深圳海思安生物技术有限公司 Cell isolation method
CN114908033A (en) * 2022-04-18 2022-08-16 广东莱迪生物医药研究院有限公司 Method for sorting cells by virtue of large-batch magnetic beads

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