CN106179544B - Portable immunomagnetic beads three-dimensional hybrid device and application method based on micro-fluidic chip - Google Patents
Portable immunomagnetic beads three-dimensional hybrid device and application method based on micro-fluidic chip Download PDFInfo
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- CN106179544B CN106179544B CN201610554419.0A CN201610554419A CN106179544B CN 106179544 B CN106179544 B CN 106179544B CN 201610554419 A CN201610554419 A CN 201610554419A CN 106179544 B CN106179544 B CN 106179544B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/12—Specific details about manufacturing devices
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Abstract
The invention discloses a kind of portable immunomagnetic beads three-dimensional hybrid devices and application method based on micro-fluidic chip, the present apparatus includes corresponding two plectanes being arranged in parallel, with extend vertically through in the shaft of two plectanes, the shaft is passed through in the position for deviateing the center of circle on plectane；Several magnets are respectively arranged on the plectane；The magnet is uniformly arranged on each plectane, and the magnet is interlaced on upright position, device of the present invention can be such that entire immunomagnetic beads mixed process carries out at ambient temperature, and whole process is automatically performed, and it is easily integrated other operating units, be conducive to further build real-time live analysis detecting system, have wide practical use.
Description
Technical field
It is particularly a kind of based on micro-fluidic chip the present invention relates to a kind of portable immunomagnetic beads three-dimensional micromixing technology
Portable immunomagnetic beads three-dimensional hybrid device, which is particularly suitable for structure real-time live analysis detecting system, and can apply
In particular surroundings.
Background technology
Immunomagnetic beads (immunomagnetic bead, IMB) technology：It is a kind of using special antigen-antibody reaction as base
The immunology detection and isolation technics of plinth.It is using the coated magnetic bead of antibody as carrier, by special in antibody and reaction medium
Property antigen binding, formed antigen-antibody complex, displacement occurs under the action of externally-applied magnetic field for this compound, so as to reach
To the purpose of separation antigen.It by solidified reagents the advantages of peculiar and the high degree of specificity of immunological response is incorporated into one, with
Based on immunology, obtained increasingly in immune detection, cell separation, biological macromolecule purifying and molecular biology etc.
It is widely applied.It, can be with being connected with the specific single of magnetic bead based on cell surface antigen by taking immunomagnetic beads cell as an example
Anti- to be combined, in externally-applied magnetic field, the cell for expressing specific antigen is connected to be adsorbed by magnetic field with magnetic bead by antibody, does not express
The cell of this kind of antigen is not combined then with immunomagnetic beads because without being adsorbed by magnetic field, buffered liquid is rinsed so as to obtain different cells
With separation.It is efficiently mixed with cell using immunomagnetic beads cell requirement immunomagnetic beads, traditional method is usually
Using gyroscope mixer, cell is made to be come into full contact with magnetic bead, so as to fulfill the specific binding of cell and magnetic bead.Gyroscope is mixed
Although clutch mixing efficiency is higher, its volume is larger, and can only generally be completed in centrifuge tube, it is difficult to realize micromation and with
Other functional units are integrated, are not suitable for structure real-time live analysis and detection device.
Invention content
To solve the above problems, a kind of portable immunomagnetic beads three-dimensional hybrid dress based on micro-fluidic chip of present invention design
It puts, which is particularly suitable for structure real-time live analysis and detection device, and can be applied to particular surroundings.
Portable immunomagnetic beads three-dimensional hybrid device of the present invention based on micro-fluidic chip, specifically, it includes
It corresponding two plectanes being arranged in parallel and extends vertically through in the shaft of two plectanes, the shaft is in stray circle on plectane
The position of the heart passes through；Several magnets are respectively arranged on the plectane；The magnet is uniformly arranged on each plectane, and institute
It is interlaced on upright position to state magnet.
Specifically, in preferred scheme, the portable immunomagnetic beads three-dimensional hybrid device, magnet is embedded in plectane
In, it uniformly arranges at each plectane upper edge edge, and interlaced on upright position.Just since shaft is in stray circle on plectane
The position of the heart passes through, and magnet is uniformly arranged at each plectane upper edge edge, and therefore, the eccentricity of each magnet is all different；In circle
In plate rotary course, due to the difference of each magnet eccentricity, cause in the mixing pit in field region and generate circulation change
Magnetic field, and then cause immunomagnetic beads in magnetic field along the movement that X-axis and Y-axis are reciprocal；Simultaneously as the magnet on two plectanes
The interlaced arrangement mode on upright position during shaft drives two plectane rotations, causes immunomagnetic beads to exist
It is moved back and forth in magnetic field along in Z-direction.Thus reach immunomagnetic beads in X-axis, Y-axis and Z axis three-dimensional motion, realization efficiently mixing
Purpose.
Specifically, in preferred scheme, the portable immunomagnetic beads three-dimensional hybrid device is further included and is matched with shaft
The driving motor and shaft base of conjunction.
Specifically, in preferred scheme, the portable immunomagnetic beads three-dimensional hybrid device further includes to fix
The clip of micro-fluidic chip.
Moreover, it relates to a kind of immunomagnetic beads three-dimensional hybrid method based on portable micro fluidic device, including
Following steps：
1. sample and immunomagnetic beads are added in the mixing pit of micro-fluidic chip；
2. the mixing pit of above-mentioned micro-fluidic chip is placed in the portable immune magnetic described above based on micro-fluidic chip
In the field region of pearl three-dimensional hybrid device；
3. two plectanes is driven to rotate by drive shaft, make to be embedded in the magnet in plectane and generate periodic Three-Dimensional Magnetic
, so as to which the immunomagnetic beads in mixing pit be driven to carry out three-dimensional motion, realize efficiently mixing.With routine immunization magnetic bead mixed method
It compares, the invention has the advantages that：
1st, using micro-fluidic chip as immunomagnetic beads three-dimensional microring array platform, which fills the present invention
It puts with small, light-weight, easy to carry, cheap, can be held.
2nd, the present invention makes entire immunomagnetic beads mixed process can by the method for microflow control technique and electromechanical integration equipment
To carry out at ambient temperature, and whole-course automation, and other operating units are easily integrated, further to build real-time live point
Analyse detecting system.
Description of the drawings
Fig. 1 is the structure diagram of the portable immunomagnetic beads three-dimensional hybrid device based on micro-fluidic chip；
Fig. 2 is micro-fluidic chip vertical view；
Fig. 3 Fig. 4 is the portable immunomagnetic beads three-dimensional hybrid principle of device schematic diagram based on micro-fluidic chip；
Fig. 5 is capture rate of the various sizes of immunomagnetic beads to CD4 lymphocytes.
In figure：Plectane on 1.；Plectane magnet on 11.；2. lower plectane；21. lower plectane magnet；3. shaft；31. rotating shaft bottom
Seat；32. driving motor；4. chip.
Specific embodiment
Following non-limiting examples can make those of ordinary skill in the art that the utility model be more fully understood, but
The utility model is not limited in any way.In the schematic diagram of device described in the utility model, structure size does not mark, mainly
The structure construction of the present apparatus is embodied, during actual production use, structure proportion and size can be adjusted as needed.
The invention will be further described with example below in conjunction with the accompanying drawings.Fig. 1 is that portable based on micro-fluidic chip is exempted from
The structure diagram of epidemic disease magnetic bead three-dimensional hybrid device, as seen from Figure 1, device of the present invention is by upper plectane 1, upper plectane magnet
11st, lower plectane 2, lower plectane magnet 21, shaft 3, shaft base 31, driving motor 32 and chip 4 form.Specifically, upper plectane 1
With lower plectane 2 is corresponding is arranged in parallel, shaft 3 is extended vertically through in two plectanes, and the shaft 3 is in deviateing the center of circle on plectane
Position passes through；Four magnets are respectively arranged on the upper plectane 1 and lower plectane 2；The magnet is uniformly arranged on each plectane
Cloth, and upper plectane magnet 11 and lower plectane magnet 21 are interlaced on upright position.
Wherein, a diameter of 50mm of two plectanes, thickness 3mm, the eccentricity of shaft 3 is 7mm, upper plectane 1 and lower circle
Include the cuboid Nd-Fe-B permanent magnet of 4 identical, symmetrical long 15mm of shape, width 6mm, high 3mm on plate 2.
It further includes the driving electricity 32 of drive shaft rotation and shaft base 31.The driving motor 32 is by minisize dc
Motor and 3V power supplys composition；
Upper plectane 1 and lower 2 horizontal parallel of plectane are placed, and upper plectane magnet 11 and lower plectane magnet 21 are staggered successively, plectane
It is driven by shaft 3 by the DC micromotor of driving motor 32；
Specifically, in preferred scheme, the portable immunomagnetic beads three-dimensional hybrid device further includes to fix
The clip of micro-fluidic chip.
Fig. 2 is micro-fluidic chip vertical view, in this micro-fluidic chip, for mixed immunity magnetic bead mixing pit in the present invention
Within the magnetic field Mixed Zone of described device.Micro-fluidic chip used in the embodiment of the present invention, mixing pit are located at
One end of micro-fluidic chip is enclosed construction, and shape, length and depth are unlimited, at least one entrance and one outlet；It will dress
The mixing pit for having the micro-fluidic chip of sample and immunomagnetic beads is positioned between the upper plectane 1 of device of the present invention, lower plectane 2
Field region in, after starting driving motor 32, upper plectane 1, lower plectane 23 start to rotate around the shaft simultaneously, when upper plectane magnet
11 when turning to Fig. 3 positions, and mixing pit top does not have magnet at this time, therefore magnetic bead is gathered in mixing pit bottom；When plectane goes to 1/8
Occurs upper plectane magnet 11 during period position, above mixing pit, lower section does not have magnet, and 1/ is gone to from initial position in two plectanes
During 8 positions, immunomagnetic beads are moved along x-axis forward direction, z-axis forward direction, while are first moved along y-axis negative sense, further along y-axis just
To movement, immunomagnetic beads are moved to the lower position of the upper plectane magnet 11 of 1/8 period position；When two plectanes are gone to 1/4 week
During the phase, lower plectane magnet 21 is located at the underface (also being located at mixing pit center) of mixing pit, and upper plectane does not have magnet position
In the surface of mixing pit, plectane from 1/8 period position during 1/4 period position, immunomagnetic beads are along x-axis is positive, z
Axis negative direction moves, while movement is that first astern negative sense, final immunomagnetic beads are gathered in mixing pit center bottom on the y axis
Portion moves；When two plectanes went to for 3/8 period, upper plectane magnet 11 is located at the surface of mixing pit, and lower plectane 2 does not have magnetic
Iron is located at the underface of mixing pit, is gone in 3/8 period position journey by 1/4 period in plectane, and immunomagnetic beads are along x-axis forward direction, z
Axis positive direction, while movement is positive movement after first negative sense on the y axis, immunomagnetic beads are moved to the upper plectane of 3/8 period position
11 lower position of magnet；When two plectanes went to for 1/2 period, the underface that lower plectane magnet 21 is located at mixing pit (also is located at
Mixing pit right end), and upper plectane 1 is located at the surface of mixing pit without magnet, and 1/2 is gone to from 3/8 period position in plectane
During period position, magnetic bead is along x-axis forward direction, z-axis negative direction, while movement is that first astern negative sense, magnetic bead gather on the y axis
Collect the other end bottom movement of mixing pit.By 1/2 period immunomagnetic beads from mixing pit one end of micro-fluidic chip along class
For spiral motion to the other end of mixing pit, movement locus is as shown in Figure 4.When the rotation of two plectanes after 1/2 period, magnet
Relative position variation with mixing pit is with preceding 1/2 period on the contrary, the sequence that i.e. Fig. 3 turns left from the right side, movement locus is also Fig. 4's
Negative direction, final magnetic bead are returned to Fig. 3 initial positions.
When two plectanes ceaselessly rotate, corresponding to the immunomagnetic beads and sample of magnetic field Mixed Zone, the transformation in magnetic field
Under, above-mentioned movement locus is constantly repeated, has been finally reached sufficient contact, mixing.
Device using the present invention and various sizes of immunomagnetic ca pture human body CD4 lymphocytes, the mixed process
It is automatically performed on micro-fluidic chip, total time is 20 minutes, is 5 in the ratio of magnetic bead and cell number:When 1, cell is put down
Equal capture rate is more than 65%, and the results are shown in Figure 5.
Claims (4)
1. a kind of portable immunomagnetic beads three-dimensional hybrid device based on micro-fluidic chip, it is characterised in that：Including corresponding
It two plectanes being arranged in parallel and extends vertically through in the shaft of two plectanes, the shaft is in the position for deviateing the center of circle on plectane
It passes through；Several magnets are respectively arranged on the plectane；Magnet is embedded in plectane, and the magnet is at each plectane upper edge edge
Uniformly arrangement, and the magnet is interlaced on upright position.
2. portable immunomagnetic beads three-dimensional hybrid device according to claim 1, it is characterised in that：It further includes and matches with shaft
The driving motor and shaft base of conjunction.
3. portable immunomagnetic beads three-dimensional hybrid device according to claim 1, it is characterised in that：It further includes to fix
The clip of micro-fluidic chip.
4. the application method of device as described in claim 1, it is characterised in that：Include the following steps,
1. sample and immunomagnetic beads are added in the mixing pit of micro-fluidic chip；
2. the mixing pit of above-mentioned micro-fluidic chip is placed in the field region of claim 1 described device；
3. two plectanes is driven to rotate by drive shaft, make to be embedded in the magnet in plectane and generate periodic three-dimensional magnetic field,
So as to which the immunomagnetic beads in mixing pit be driven to carry out three-dimensional motion, efficiently mixing is realized.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610554419.0A CN106179544B (en) | 2016-07-14 | 2016-07-14 | Portable immunomagnetic beads three-dimensional hybrid device and application method based on micro-fluidic chip |
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| CN201610554419.0A CN106179544B (en) | 2016-07-14 | 2016-07-14 | Portable immunomagnetic beads three-dimensional hybrid device and application method based on micro-fluidic chip |
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| CN106179544A CN106179544A (en) | 2016-12-07 |
| CN106179544B true CN106179544B (en) | 2018-07-06 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109718873A (en) * | 2017-10-31 | 2019-05-07 | 中国科学院大连化学物理研究所 | The polynary immune response system of micro- magnetic bead based on digital drop micro-fluidic chip |
| CN112213174A (en) * | 2019-07-12 | 2021-01-12 | 南京岚煜生物科技有限公司 | Method for uniformly mixing magnetic materials |
| CN110938522A (en) * | 2019-12-13 | 2020-03-31 | 大连海事大学 | Portable bacteria cracking device based on centrifugal microfluidic technology and use method thereof |
| CN110947438B (en) * | 2019-12-13 | 2022-05-24 | 大连海事大学 | Three-dimensional magnetic field parallel mixing microfluidic device and using method thereof |
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