CN107621486A - The program control layout inoculation method of cell and device of Cell-oriented impedance detection research - Google Patents
The program control layout inoculation method of cell and device of Cell-oriented impedance detection research Download PDFInfo
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- CN107621486A CN107621486A CN201710673793.7A CN201710673793A CN107621486A CN 107621486 A CN107621486 A CN 107621486A CN 201710673793 A CN201710673793 A CN 201710673793A CN 107621486 A CN107621486 A CN 107621486A
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Abstract
The present invention discloses a kind of program control layout inoculation method of cell of Cell-oriented impedance detection research and device, bottom inside cell culture chamber is equipped with interdigital electrode, interdigital electrode is drawn golden film and golden film is drawn to electrode and draws golden film with working electrode and the perpendicular N number of strip type gold electrode of golden film is drawn to electrode and formed by the working electrode being parallel to each other, equidistantly M micro-hole is provided with center line between each strip type gold electrode both ends, it is a micro- magnetic pole immediately below each micro-hole, forms the micro- array of magnetic dipoles being made up of M × N number of micro- magnetic pole；Attract the technology of the immunomagnetic beads with reference to cell with micro- magnetic pole of energization, according to the position for micro- magnetic pole that is powered in the advance micro- array of magnetic dipoles of cell inoculation layout determination, position and the number for micro- magnetic pole that is powered in micro- array of magnetic dipoles are automatically determined by stored program control system, make the layout of the cell distribution after inoculation identical with target, realize the inoculation of program control layout and automation inoculation of cell.
Description
Technical field
The invention belongs to cell impedance detection field, refers in particular to a kind of program control layout inoculation method of cell and device.
Background technology
Cell impedance detection technique has been widely used in the research of cell biology, and cell attachment is when interdigital electrode
Impedance detection can effectively evaluate the active change of cell, the proliferative conditions of cell, the migration velocity of cell, cell it is anti-
Malicious ability etc..However, in cell impedance detection process is carried out with interdigital electrode, cell is in interdigital electrode cell sensor table
The distribution situation in face（The uniformity of such as distribution, the position of distribution, the dense degree of distribution）With the precision of cell impedance detection
It is closely related.For a variety of layout inoculation methods of cell, that currently mainly studies has antibody inocalation method, template inocalation method etc..
Antibody inocalation method therein is mainly the method combined using cell with antibody specificity, and being inoculated with cell in chamber needs inoculation thin
On the diverse location of born of the same parents, manually antibody is applied on different location points under the microscope using miniature dropper, then by cell
In buffer solution injection inoculation chamber, because antibody is combined with the characteristic of cell, cell can move towards the direction for having antibody, so that
Cell attaches in specific position.But this manually-operated technical requirements of method are higher, it is necessary to operated under the microscope,
Operation difficulty is larger, and manual operation error is larger in the coating process of antibody, and then is forbidden the inoculation position of cell
Really.Template inocalation method therein is that processing forms miniature hole array on thin-film template in advance, and the position in each hole represents institute
Want the position of inoculating cell.Thin-film template is placed into cell inoculation chamber, then injected cell culture fluid in inoculation chamber.When
During cell precipitation, porose band of position cell can be attached at the bottom of inoculation chamber in template, and be not in the thin of bore region
Born of the same parents are only attached in template.Finally thin-film template is removed, leaves the cell for being seeded in inoculation bottom of chamber portion specific region.Template connects
Kind method needs the template that pre-production is inoculated with, and the inoculation layout of template can not be changed, can not again dynamically in seeded process
Inoculation layout of the ground to cell is adjusted and changed, since it is desired that manually paving template is with going template and being unable to reach automatic vaccination
Requirement.Therefore, research process are detected in cell impedance, can not be to entering with a collection of cell under different distributions layout state
Row research, extends research cycle.
The content of the invention
The shortcomings that the invention aims to overcome above cell inoculation method, propose a kind of Cell-oriented impedance detection
The program control layout inoculation method of cell and device of research, the requirement of layout change is inoculated with dynamically to cell point according to cell at any time
Cloth layout is adjusted change, improves the progress of research, and the impedance under different distributions state with a collection of cell can be carried out
Continuous detection.
The technical scheme that the program control layout classification inoculation apparatus of cell of Cell-oriented impedance detection research of the present invention uses
It is：Including electric cell-substrate impedance sensor, electric cell-substrate impedance sensor top is cell culture chamber, and the bottom inside cell culture chamber is equipped with
Interdigital electrode, interdigital electrode draws golden film by a working electrode being parallel to each other and one is drawn golden film and and work to electrode
Make electrode to draw golden film and draw electrode the perpendicular N number of strip type gold electrode composition of golden film, the bar of interlaced half
Shape type gold electrode one end connection working electrode draws golden film, and second half interlaced strip type gold electrode one end is connected to electricity
Draw golden film in pole；M micro-hole equidistantly is provided with center line between each strip type gold electrode both ends, each micro-hole is just
Lower section is a micro- magnetic pole, forms the micro- array of magnetic dipoles being made up of M × N number of micro- magnetic pole；It is one thin at the top of each micro- magnetic pole
Needle, an electromagnetic core of every thin needle connection bottom, one spiral winding of each electromagnetic core outside winding；Each spiral shell
Spin line circle connects embedded microcontroller through micro- array of magnetic dipoles control module, and working electrode draws golden film and draws gold to electrode
Film connects embedded microcontroller through cell impedance detection module.
Further, electric cell-substrate impedance sensor is embedded in a groove alloy block inside, and groove alloy block is fixed and is located at
On the upper surface of rocking platform, rocking platform is driven by steering wheel to be vibrated up and down, and steering wheel declines through the connection insertion of servo driving module
Controller.
The inoculation method of the program control layout classification inoculation apparatus of cell of Cell-oriented impedance detection research of the present invention uses
Technical scheme be to comprise the following steps：
A, will combine cell magnetic bead buffer solution injection cell culture chamber in, while by cell inoculation layout be built in it is embedded
In microcontroller；
B, embedded microcontroller is inoculated with layout according to cell, to each micro- since the first row first row of micro- array of magnetic dipoles
Whether cell vaccination needs inoculating cell to be judged corresponding to magnetic pole, is if desired inoculated with, and drives the helix of micro- magnetic pole
Circle is powered, and makes micro- magnetic pole of the position that magnetic be presented, the magnetic thin needle of tool attracts the magnetic bead with reference to cell, and magnetic bead is attached at
Thin needle upper end, have distribution of the magnetic micro- magnetic pole in whole array with being needed in cell inoculation layout in micro- array of magnetic dipoles
Want the distributing position of the point of inoculating cell identical；
C, after needing the vaccination of inoculating cell to have cell attachment, embedded microcontroller control cell impedance detection mould
Block is worked, and ac voltage signal is applied into working electrode extraction golden film and electrode is drawn in golden film, the current signal of backflow
To embedded microcontroller, embedded microcontroller calculates cell impedance value for input after cell impedance detection module.
Further, in step B, embedded microcontroller swings back and forth through servo driving module control steering wheel, steering wheel band
Electric cell-substrate impedance sensor on dynamic rocking platform makees low frequency micro oscillation.
It is of the invention compared with existing methods and techniques, have the following advantages that：
1st, during research cell distribution and cell impedance accuracy of detection relation, the present invention attracts with the micro- magnetic pole being powered
With reference to the technology of the immunomagnetic beads of cell, according to the position for micro- magnetic pole that is powered in the advance micro- array of magnetic dipoles of cell inoculation layout determination
Put, position and the number of micro- magnetic pole that is powered in micro- array of magnetic dipoles are automatically determined by stored program control system, makes electric cell-substrate impedance sensor
In the position of cell towards the micro- magnetic pole of being powered attached, the cell distribution after inoculation is laid out and target phase
Together, the program control layout inoculation of cell is realized, the automation inoculation of cell is realized, eliminates cumbersome manual operation process, drop
The low experimental implementation technical difficulty of experimenter,
2nd, the present invention is compared with the method that manual type in antibody inocalation method establishes cell inoculation layout, when overcoming manual layout
The shortcomings that position error is big, there is the characteristics of automation inoculation, the accuracy of cell inoculation cloth office point position can be greatly improved.
3rd, the present invention can be inoculated with wanting for layout change according to cell at any time during cell impedance accuracy of detection is studied
Ask and change dynamically is adjusted to cell inoculation layout, compared with template inocalation method, overcome template inocalation method inoculation layout
The shortcomings that fixed single, there is great flexibility and convenience.
4th, the present invention makes cushioning liquid do the reciprocating motion of low frequency using mechanical micro oscillation, and band kinetocyte and magnetic bead combine
Body moves, and ensure that all cells-magnetic bead combination has the chance in micro- magnetic pole attractived region, avoids micro- magnetic indirectly
Pole attracts the generation of blind area.
Brief description of the drawings
Fig. 1 is the structural representation of the program control layout classification inoculation apparatus of the cell of Cell-oriented impedance detection research of the present invention
Figure；
Fig. 2 is the stereochemical structure enlarged drawing of electric cell-substrate impedance sensor in Fig. 1；
Fig. 3 is Fig. 1 controling circuit structure block diagram；
Fig. 4 is micro- array of magnetic dipoles distribution and cell inoculation layout corresponding relation figure in Fig. 2；
The sequence number and title of each part in accompanying drawing：1. cell culture chamber；2. strip type gold electrode；3. working electrode draws golden film；
4. pair electrode draws golden film；5. micro-hole；6. thin needle；7. electromagnetic core；8. spiral winding；9. micro- magnetic pole；10. connecting pole；
11. sensor base plate；12. micro- array of magnetic dipoles power supply interface；13. cell impedance detects port；14. electric cell-substrate impedance sensor；15.
Rocking platform；16. groove alloy block；17. connecting bracket；18. steering wheel；19. steering wheel support；20. steering wheel cantilever；21. system control
Box processed；22. support spring；24. fixing bolt.
Embodiment
Referring to Fig. 1, the program control layout classification inoculation apparatus of cell of Cell-oriented impedance detection research of the present invention includes one
There is program control layout to be inoculated with function for individual electric cell-substrate impedance sensor 14, electric cell-substrate impedance sensor 14, and electric cell-substrate impedance sensor 14 is just
It is embedded in groove alloy block 16, groove alloy block 16 is fixed on the upper surface of rocking platform 15, and rocking platform 15 is by steering wheel 18
Drive and vibrate up and down.
On the upper surface of rocking platform 15, a connecting bracket 17 is fixed positioned at the outside of groove alloy block 16 side, even
The lower end that support 17 is fixedly connected with steering wheel cantilever 20 by fixing bolt 24 is connect, steering wheel cantilever 20 is arranged vertically, and its upper end is fixed
The output shaft of steering wheel 18 is connected, the output shaft of steering wheel 18 is horizontally disposed, drives steering wheel cantilever 20 to swing back and forth.Rocking platform 15
Lower surface connects support spring 22, is supported on the ground by support spring 22.When steering wheel 18 swings back and forth, in support spring
In the presence of 22, connecting bracket 17 is driven to promote rocking platform to be vibrated about 15 by steering wheel cantilever 20.The casing of steering wheel 18 passes through
Steering wheel support 19 fixes on the ground.Steering wheel support 19 is fixedly connected with the casing of steering wheel 18 perpendicular to ground, its upper end, and lower end is solid
It is scheduled on ground.
Electric cell-substrate impedance sensor 14 and steering wheel 18 are respectively connected to inside system control box 21 by signal wire, system control box
21 inside have embedded microcontroller, and embedded microcontroller controls electric cell-substrate impedance sensor 14 to carry out the program control cloth of cell respectively
Office and cell impedance detection, and control the swing of steering wheel 18.
The structure of electric cell-substrate impedance sensor 14 shown in Figure 2, its top are one square made of quartz glass
Cell culture chamber 1, using magnetron sputtering technology, an interdigital electrode, the fork are installed in the bottom inside cell culture chamber 1
Refer to electrode be by working electrode draw golden film 3, one electrode is drawn golden film 4 and the width formed according to certain intervals and
Same N number of strip type gold electrode 2 composition of identical length, working electrode draws golden film 3 and draws golden film 4 to electrode and is parallel to each other
Arrangement, it is to draw golden film 3 with working electrode and the perpendicular N number of strip type gold electrode 2 of golden film 4 is drawn to electrode between the two.N
One end connection working electrode of the strip type gold electrode 2 of interlaced half in individual strip type gold electrode 2 draws golden film 3,
The other end is pointed to but do not contacted draws golden film 4 to electrode.Second half interlaced one end of strip type gold electrode 2 is connected to electricity
Golden film 4 is drawn in pole, and the other end, which points to but do not contact working electrode, draws golden film 3.
On center line between the both ends of each strip type gold electrode 2, according to equidistant principle, by cheesing techniques,
Each to beat M micro-hole 5, common M × N number of micro-hole 5, the position where each micro-hole 5 is exactly each cell vaccination.Every
The underface of individual micro-hole 5 is a micro- magnetic pole 9, so just forms a micro- magnetic pole battle array being made up of M × N number of micro- magnetic pole 9
Row.Micro- magnetic pole 9 is collectively constituted by thin needle 6, electromagnetic core 7 and spiral winding 8.The top of each micro- magnetic pole 9 is a thin magnetic
Pin 6, every thin needle 6 connect an electromagnetic core 7 of bottom, one spiral winding 8 of outside winding of electromagnetic core 7, spiral
Coil 8 is wound in the surface of electromagnetic core 7 by enamel-covered wire according to a certain direction.Electromagnetic core 7 is located at outside cell culture chamber 1
Lower section, thin needle 6 is from bottom to up through corresponding aperture on the bottom surface of cell culture chamber 1, and be inserted into each micro-hole 5
In, and the top of thin needle 6 is equal with bottom cell culture chamber 1 inside, then is filled in thin needle 6 and carefully with nontoxic marine glue
The gap of born of the same parents' culture chamber 1 and micro-hole 5, thin needle 6 is fixedly connected on micro-hole 5, also make thin needle 6 and cell culture chamber 1
Between seal.The material of thin needle 6 is the permalloy for having high permeability, and the most thin section diameter in its upper end is micron order,
This is suitable with the size of most cells.
The lower end of electromagnetic core 7 is fixed on sensor base plate 11, and sensor base plate 11 is horizontally disposed.Cell culture chamber 1
Four angles of outer bottom are connected on sensor base plate 11 by a connecting pole 10, the length of connecting pole 10 and each micro- magnetic
Pole 9 is contour, and plays a part of fixed and support.
Each spiral winding 8 has two terminals, and one end is that electric current flows into end, and the other end is electric current outflow end.Per opposite end
Son collects to form bus by wire, is connected on micro- array of magnetic dipoles power supply interface 12, and micro- array of magnetic dipoles power supply interface 12 is solid
It is scheduled on sensor base plate 11.Working electrode, which draws golden film 3 and golden film 4 is drawn to electrode, is wired to cell impedance inspection
Survey on port 13, cell impedance detection is fixed on sensor base plate 11 on port 13.
The outward flange of sensor base plate 11 and the fit depressions of groove alloy block 16, sensor base plate 11 close embedded in groove
In gold bullion 16.
Referring to Fig. 3, inside system control box 21 in addition to embedded microcontroller, also controlled with micro- array of magnetic dipoles
Module, cell impedance detection module and servo driving module.Embedded microcontroller is connected respectively by different control ports
Micro- array of magnetic dipoles control module, cell impedance detection module and servo driving module.Micro- array of magnetic dipoles control module is through micro- magnetic pole
Powering arrays interface 12 connects and drives each spiral winding 8, and cell impedance detection module connects through cell impedance detection port 13
Connect and control working electrode to draw golden film 3 and golden film 4 is drawn to electrode, embedded microcontroller control cell impedance detection module
Detect port 13 by cell impedance and draw golden film 3 in working electrode and electrode is drawn and apply ac voltage signal in golden film 4,
And calculate cell impedance after gathering the current signal of backflow.Servo driving module connects and drives steering wheel 18.Wherein, micro- magnetic pole
Array control module is made up of a multi-thread decoder, M × N number of output latch, M × N number of coil drive module.Micro- magnetic pole battle array
Row control module is connected by universal input/output interface of multi-thread a decoder and embedded microcontroller of inside, more
The output end connection of line decoder and the quantity identical output latch of micro- magnetic pole 9, each output latch connect a line
The switch port of drive module is enclosed, passes through the opening and turning off come control coil drive module of the output signal of output latch.
Low-voltage, high-current voltage-stabilized power supply is connected with the power supply terminal of each coil drive module.The drive of each coil drive module
Moved end mouth is pooled to micro- pole coil driving parallel interface input side with bus mode, i.e., shared M × N number of coil drive module
Driving port is connected to micro- pole coil driving parallel interface input side.Micro- pole coil driving parallel interface outlet side and input
Side corresponds, and is connected on micro- array of magnetic dipoles power supply interface 12 shown in Fig. 2, makes each coil drive module one
An one correspondingly connected spiral winding 8., it is necessary to be inoculated with after embedded microcontroller exports the control signal of micro- array of magnetic dipoles
There is electric current in micro- magnetic pole spiral winding 8 under cell compartment by the magnetic line of force being produced in corresponding electromagnetic core 7, so as to thin magnetic
Pin 6 is magnetized, and can attract to be combined with the immunomagnetic beads of cell, cell can be with position attaching.
There is corresponding relation in the distribution for the micro- array of magnetic dipoles being made up of M × N number of micro- magnetic pole 9, corresponding with cell inoculation layout
Graph of a relation in Fig. 4 by taking micro- array of magnetic dipoles of M=N=8 as an example as shown in figure 4, only explain.Each small square generation in Fig. 4
One cell vaccination of table, the position corresponding to the micro-hole 5 of the position correspondence of each cell vaccination in Fig. 1, cell
Vaccination bottom is a micro- magnetic pole 9, i.e. each cell vaccination is corresponding with each micro- magnetic pole 9.The small pros of white
Shape represents the no power of micro- magnetic pole 9 of the vaccination not inoculating cell, i.e. the vaccination bottom, nonmagnetic, it is impossible to which attraction is combined with
The immunomagnetic beads of cell；The small square of black represents that the vaccination needs micro- magnetic pole 9 of inoculating cell, i.e. the vaccination bottom
It is powered, is magnetic, can attract to be combined with the immunomagnetic beads of cell.Cell impedance accuracy of detection is influenceed in research cell distribution
During, according to the cell requirement that diverse location is distributed on electrode, by changing position and the quantity of vaccination, can be formed
A variety of cells are inoculated with layout.Cell vaccination position distribution as shown according to Fig. 4, it can be formed a kind of specific
Cell is inoculated with layout.
Referring to Fig. 1-4, the course of work of the program control layout inoculation method of cell of Cell-oriented impedance detection research of the present invention
It is specific as follows：
It is first to prepare for work：It is put into jointly with the cell to be inoculated with by the magnetic bead that surface is manually surrounded by specific antibody
In buffer solution, after a period of time, the cell to be inoculated with produces specific binding with immunomagnetic beads, just form with reference to needed for
Want the magnetic bead cushioning liquid of inoculating cell.The cell culture of the magnetic bead buffer solution injection electric cell-substrate impedance sensor 14 of cell will be combined
In chamber 1, then electric cell-substrate impedance sensor 14 is embedded in groove alloy block 16.Cell inoculation layout is built in simultaneously embedding
Enter in the controller that declines.
Embedded microcontroller is inoculated with layout according to cell, and the of the micro- array of magnetic dipoles formed from the micro- magnetic pole 9 of M × N number of
Whether a line first row starts, need inoculating cell to judge cell vaccination corresponding to each micro- magnetic pole 9, if desired connect
The micro- magnetic pole 9 for planting the then position correspondence should have magnetic, then drive the coil drive module of micro- magnetic pole 9 open-minded, i.e., embedded
Controller decline to driving the coil drive module of each micro- magnetic pole 9（I.e. one shared M × N number of coil drive module）It is open-minded
Judged with shut-off.The i-th row jth row spiral shell of the coil drive module array formed for M × N number of coil drive module
Spin line circle 8, its break-make represent to minor function：
,
Wherein,.After the completion of each coil drive module and the break-make of spiral winding 8 judge, just
Obtain the encoder matrix A of M rows N row, the value of each element in encoder matrix A is 1 or 0, represents correspondence position
Coil drive module and spiral winding 8 being opened and turning off.
The encoded radio that cell is inoculated with the encoder matrix A of layout by embedded microcontroller is sent into multi-thread decoder, multi-thread to translate
Code device just obtains the driving control signal of each micro- magnetic pole 9 in micro- array of magnetic dipoles after decoding.By output latch, need
Relay conducting in coil drive module corresponding to the micro- magnetic pole 9 to be opened, so that required inoculating cell region bottom
The spiral winding 8 of micro- magnetic pole 9 be powered, make micro- magnetic pole 9 of the position that magnetic be presented, so that it is thin to be connected with micro- magnetic pole 9
Needle 6 is magnetized rapidly, is had distribution of the magnetic micro- magnetic pole 9 in whole array in micro- array of magnetic dipoles and is laid out with cell inoculation
Need the distributing position of the point of inoculating cell identical in figure.So, the thin needle 6 for having magnetic micro- top of magnetic pole 9 can attract
The neighbouring magnetic bead for combining cell, it is allowed to be adsorbed on the upper end of thin needle 6.Now, the magnetic in the cushioning liquid in culture chamber
Pearl has three kinds of different states：A part is combined with the magnetic bead of cell due to being adsorbed on close to the thin needle 6 being magnetized
On thin needle 6, the thin needle 6 that the magnetic bead of another part combination cell is magnetized due to distance farther out, between thin needle 6 and magnetic bead
Caused attraction be not enough to overcome buffer solution to the resistance of magnetic bead and in state is not attracted, also sub-fraction magnetic bead by
Magnetic bead has been adsorbed in the thin needle 6 being nearby magnetized and has been free near it, so this fashion has part to combine cell
Magnetic bead does not reach vaccination due to attracting blind area not to be attracted.
In order that the position of inoculating cell in need have a cell, the present invention solves attraction blind area using rocking platform 15
The problem of.Micro- magnetic pole 9 below inoculating cell is needed in micro- array of magnetic dipoles, and by after driving simultaneously, embedded microcontroller exports
PWM ripples control the steering wheel cantilever 20 of steering wheel 18 to swing back and forth once through servo driving module, and steering wheel cantilever 20 is to rocking platform 15
Produce and extrude, the support spring 22 under rocking platform 15 is accordingly extruded, and when 20 return to the nature state of steering wheel cantilever, supports bullet
Spring 22 produces damped oscillation, drives the electric cell-substrate impedance sensor 14 on rocking platform 15 to make low frequency micro oscillation.In low frequency micro oscillation
During, the magnetic bead for being combined with cell not being attracted can correspondingly produce motion, when moving to also unadsorbed magnetic bead and tool
Then it is attracted when near magnetic thin needle 6, and the thin needle 6 for having adsorbed magnetic bead will not then produce motion and keep
In current position.After stopping is vibrated, having magnetic position has magnetic bead absorption, i.e., the vaccination of required inoculating cell
Has cell attachment.
After the vaccination of required inoculating cell has cell attachment, whole device is stood into a period of time, it is embedded
The controller that declines, which is sent, to be started order and cell impedance detection module is worked, and cell impedance detection produces certain frequency and amplitude
Ac voltage signal, and port 13 is detected by cell impedance and is applied to working electrode extraction golden film 3 and to electrode extraction golden film 4
On, the current signal of backflow amplifies with after filtering, being finally-transmitted to embedded microcontroller by cell impedance detection module.So
Embedded microcontroller is according to the ac voltage signal amplitude of application, frequency and the back flow current signal amplitude calculating collected afterwards
Go out cell impedance value now.
After the cell impedance that layout is inoculated with to a kind of cell is measured, if necessary to carry out second of Cell-oriented resistance
The program control layout inoculation of cell of anti-detection research, then embedded microcontroller is first turned off all micro- magnetic poles 9 in micro- array of magnetic dipoles
Coil drive module, that is, turn off all spiral windings 8, now all micro- magnetic poles 9 in micro- array of magnetic dipoles are nonmagnetic, knot
The immunomagnetic beads that closing has cell is in random distribution state.Then again by second cell be inoculated with layout be built in it is embedded
In microcontroller, embedded microcontroller starts to connect according to second of program control layout of cell inoculation layout second of cell of progress
The process of kind, second of cell impedance measurement is just carried out after the program control layout inoculation of second of cell, has so just been reached thin
The effect adjusted into Mobile state with changing is laid out in the anti-detection process of the embarrassment of fetus to cell inoculation.So circulation, until completing to institute
The inoculation for having inoculation layout detects with corresponding cell impedance.
Claims (7)
1. a kind of program control layout classification inoculation apparatus of the cell of Cell-oriented impedance detection research, including electric cell-substrate impedance sensor（14）,
Electric cell-substrate impedance sensor（14）Top is cell culture chamber（1）, it is characterized in that：Cell culture chamber（1）Internal bottom is equipped with fork
Refer to electrode, interdigital electrode draws golden film by a working electrode being parallel to each other（3）Golden film is drawn to electrode with one（4）And
Golden film 3 is drawn with working electrode and golden film is drawn to electrode（4）Perpendicular N number of strip type gold electrode（2）Composition, it is interlaced
Half strip type gold electrode（2）One end connection working electrode draws golden film（3）, interlaced second half strip type gold
Electrode（2）One end connection draws golden film to electrode（4）；In each strip type gold electrode（2）On center line between both ends equidistantly
Provided with M micro-hole（5）, each micro-hole（5）Underface is a micro- magnetic pole（9）, formed by M × N number of micro- magnetic pole（9）Composition
Micro- array of magnetic dipoles；Each micro- magnetic pole（9）Top be a thin needle（6）, every thin needle（6）Connect an electricity of bottom
Magnetic core（7）, each electromagnetic core（7）One spiral winding of outside winding（8）；Each spiral winding（8）Through micro- magnetic pole battle array
Row control module connects embedded microcontroller, and working electrode draws golden film（3）Golden film is drawn with to electrode（4）Through cell impedance
Detection module connects embedded microcontroller.
2. the program control layout classification inoculation apparatus of the cell of Cell-oriented impedance detection research according to claim 1, it is characterized in that：
Electric cell-substrate impedance sensor（14）It is embedded in a groove alloy block（16）In inside, groove alloy block（16）Fix to be located to vibrate and put down
Platform（15）Upper surface on, rocking platform（15）By steering wheel（18）Drive and vibrate up and down, steering wheel（18）Connect through servo driving module
Connect embedded microcontroller.
3. the program control layout classification inoculation apparatus of the cell of Cell-oriented impedance detection research according to claim 2, it is characterized in that：
In rocking platform（15）Upper surface on, positioned at groove alloy block（16）Fixation sets a connecting bracket beside outside（17）, even
Connect support（17）It is fixedly connected with vertically arranged steering wheel cantilever（20）Lower end, steering wheel cantilever（20）Upper end is fixedly connected horizontally disposed
Steering wheel（18）Output shaft；Rocking platform（15）Lower surface connection support spring（22）, support spring（22）It is supported on ground
On face.
4. the program control layout classification inoculation apparatus of the cell of Cell-oriented impedance detection research according to claim 1, it is characterized in that：
Micro- array of magnetic dipoles control module is made up of a multi-thread decoder, M × N number of output latch, M × N number of coil drive module, micro-
Array of magnetic dipoles control module is connected by a multi-thread decoder with embedded microcontroller, the output end connection of multi-thread decoder
Output latch, each output latch connect a coil drive module, and control coil drive module being opened and turning off, often
One coil drive module connects a spiral winding correspondingly（8）.
A kind of 5. inoculation method of the program control layout classification inoculation apparatus of cell as claimed in claim 2, it is characterized in that including following step
Suddenly：
A, the magnetic bead buffer solution for combining cell is injected into cell culture chamber（1）In, at the same by cell inoculation layout be built in it is embedding
Enter in the controller that declines；
B, embedded microcontroller is inoculated with layout according to cell, to each micro- since the first row first row of micro- array of magnetic dipoles
Magnetic pole（9）Whether corresponding cell vaccination needs inoculating cell to be judged, is if desired inoculated with, and drives micro- magnetic pole（9）'s
Spiral winding（8）It is powered, makes micro- magnetic pole of the position（9）Magnetic is presented, has magnetic thin needle（6）Attract with reference to cell
Magnetic bead, magnetic bead are attached at thin needle（6）Upper end, has magnetic micro- magnetic pole in micro- array of magnetic dipoles（9）Point in whole array
The distributing position that cloth is inoculated with the point that inoculating cell is needed in layout with cell is identical；
C, after needing the vaccination of inoculating cell to have cell attachment, embedded microcontroller control cell impedance detection mould
Block works, and ac voltage signal is applied into working electrode draws golden film（3）Golden film is drawn with to electrode（4）On, the electricity of backflow
Stream signal is inputted to embedded microcontroller, embedded microcontroller after cell impedance detection module calculates cell impedance
Value.
6. according to the inoculation method of the program control layout classification inoculation apparatus of cell as claimed in claim 5, it is characterized in that：It is embedding in step B
The controller that enters to decline controls steering wheel through servo driving module（18）Swing back and forth, steering wheel（18）Drive rocking platform（15）On it is thin
Born of the same parents' impedance transducer（14）Make low frequency micro oscillation.
7. according to the inoculation method of the program control layout classification inoculation apparatus of cell as claimed in claim 5, it is characterized in that：After step C,
After the completion of cell impedance detection to a kind of cell inoculation layout, embedded microcontroller is first turned off all spiral windings
（8）, second of cell is inoculated with layout again and is built in embedded microcontroller, then starts to connect according to second of cell
Kind of layout carries out the process of the program control layout inoculation of cell, so circulation, until complete the inoculations of all inoculation layouts with it is right
The cell impedance detection answered.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710673793.7A CN107621486B (en) | 2017-08-09 | 2017-08-09 | The program-controlled layout inoculation method of the cell of Cell-oriented impedance detection research and device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710673793.7A CN107621486B (en) | 2017-08-09 | 2017-08-09 | The program-controlled layout inoculation method of the cell of Cell-oriented impedance detection research and device |
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| Publication Number | Publication Date |
|---|---|
| CN107621486A true CN107621486A (en) | 2018-01-23 |
| CN107621486B CN107621486B (en) | 2019-12-03 |
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