CN109632612A - A kind of cell resistance anti-detection system and method simultaneously with extracellular field potential high throughput - Google Patents
A kind of cell resistance anti-detection system and method simultaneously with extracellular field potential high throughput Download PDFInfo
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- CN109632612A CN109632612A CN201910055927.8A CN201910055927A CN109632612A CN 109632612 A CN109632612 A CN 109632612A CN 201910055927 A CN201910055927 A CN 201910055927A CN 109632612 A CN109632612 A CN 109632612A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects thereof, e.g. conductivity or capacity
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/60—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrostatic variables, e.g. electrographic flaw testing
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
Abstract
The invention discloses a kind of cell resistance anti-detection system and method simultaneously with extracellular field potential high throughput, which includes that cell resistance is anti-with extracellular field potential compound sensor, the anti-conditioning circuit of cell resistance, the extracellular field potential conditioning circuit of cell and data collecting card；Compound sensor is made of porous cell culture cavity, the anti-sensor of cell resistance and the extracellular field potential sensor of cell；Porous cell culture cavity, which is fixed in substrate of glass, forms independent multiple cell culture chambers, and the bottom of each cell culture chamber is equipped with independent compound sensor unit.The present invention is selected by sample frequency and notch filter double shield, it is noiseless while detecting to realize two kinds of signals, and have the characteristics that high sensitivity, flux are high；Existing cell impedance single hole scan-type detection is overcome, especially with the deficiency of time delay in the case where high throughput；Operation of the present invention step is simple, has the advantages that continuous detection, applicable object area are extensive for a long time to cell.
Description
Technical field
The present invention relates to a kind of no-delay high-throughput detection technique of multi-parameter in cell culture more particularly to a kind of cell electricity
Impedance and extracellular field potential high throughput detection system and method simultaneously.
Background technique
Biosensor be it is a kind of by biological " element " in conjunction with physical chemistry energy converter, for analyzing the dress of analyte detection
It sets.Emerged in recent years the biological derivatives such as bio-tissue, microorganism, cell, enzyme, antibody or bionical substance in physics,
The example of the various fields such as chemistry, biology application, wherein cell sensor is the emerging life that recent decades grow up
Object analysis and detection technology, one of the important branch as biosensor pass through two using cell as primary characterization element
Stimuli responsive of the cell to identification signal is converted into light, electricity or chemistry etc. by the sensors such as grade energy converter such as physics, chemistry can
The signal of measurement.Therefore, cell sensor is since appearance, since it is with good sensitivity and specificity, it is easy to operate simultaneously
And cost is relatively low for high throughput, and by cell sensor and particular technology platform intergration, the real-time monitoring to analyte may be implemented,
It is expected to be promoted in fields such as odor identification, Pharmaceutical Analysis.
Summary of the invention
In view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to which it is anti-with extracellular field potential high pass to provide a kind of cell resistance
Amount while detection system and method.
The purpose of the present invention is what is be achieved through the following technical solutions: a kind of cell resistance is anti-high-throughput with extracellular field potential
Detection system simultaneously, comprising: cell resistance resists and extracellular field potential compound sensor, the anti-conditioning circuit of cell resistance, cell born of the same parents
Outer field potential conditioning circuit and data collecting card.
Wherein, cell resistance is anti-and extracellular field potential compound sensor is by porous cell culture cavity, the anti-biography of cell resistance
Sensor and the extracellular field potential sensor composition of cell, the anti-sensor of cell resistance are made of interdigital electrode, the extracellular field potential of cell
Sensor is made of plane electrode, and two kinds of sensor combinations are at independent compound sensor unit；Porous cell culture cavity is solid
Fixed to form independent multiple cell culture chambers on the glass substrate, the bottom of each cell culture chamber is equipped with independent multiple
Close sensor unit.
Data collecting card includes excitation generation module and signal acquisition module, and excitation generation module passes through excitation conditioning circuit
Respectively with the excitation end of the anti-sensor interdigital electrode of cell resistance in the compound sensor unit of each cell culture chamber bottom
Electrical connection, for providing ac-excited signal；The test side of interdigital electrode receives signal and passes through the anti-conditioning circuit of cell resistance
Amplify, filter after be input to the signal acquisition module of data collecting card；The extracellular field potential sensor of cell passes through cell born of the same parents
Outer field potential conditioning circuit amplifies, filter after be input to the signal acquisition module of data collecting card.
Further, which further includes power management module, for being the anti-conditioning circuit of cell resistance and cell
Extracellular field potential conditioning circuit power supply.
Further, each independent compound sensor unit includes two pairs of interdigital electrodes and two plane electrodes；Two pairs
Interdigital electrode symmetry arrangement, excitation end share, and test side is separated, exports the anti-detection signal of two-way cell resistance；Two plane electricity
Pole exports the extracellular field potential detection signal of cell all the way respectively.
Further, excitation conditioning circuit will motivate the ac-excited signal of generation module output to decay to to be checked first
The undamaged voltage range of the cell of survey, is then filtered, and filters out the excitation end that interdigital electrode is input to after noise jamming.
Further, the anti-conditioning circuit of cell resistance includes current-voltage conversion circuit, high-pass filter and voltage follow
Device, current-voltage conversion circuit are used to convert the current signal that interdigital electrode test side detects to voltage signal, high pass filter
Wave device is used for influence of the isolation signals output end to input terminal for removing baseline drift, voltage follower.
Further, the extracellular field potential conditioning circuit of cell include low noise pre-amplification circuit, trap-high-pass filter,
Two-stage amplifier and voltage follower, low noise pre-amplification circuit are used for the extracellular field potential of cell that enlarged plan electrode detection arrives
Signal is detected, trap-high-pass filter is for filtering out pumping signal and baseline drift, and two-stage amplifier is for being further amplified inspection
It surveys signal and is used for influence of the isolation signals output end to input terminal to grade, voltage follower is lied prostrate.
A kind of cell resistance is anti-with extracellular field potential high throughput Simultaneous Detection, comprising the following steps:
(1) myocardial cells culture liquid is prepared, by a certain number of transplantation of cardiomyocytes in each cell culture chamber；
(2) by data collecting card generate sinusoidal excitation signal, by motivate conditioning circuit decay to cardiac muscle cell without
The voltage range of damage is applied to the excitation end of the anti-sensor interdigital electrode of cell resistance after filtering out noise, then passes through interdigital electricity
Pole test side receives current signal, since the interdigital electrode excitation end of each independent cell culture chamber shares, test side point
It opens, forms the independent multiple measure loops in every hole, therefore the no-delay cell resistance of multichannel may be implemented and resist while detecting；
(3) while step (2) detect cell resistance antinoise signal, plane electrode records the extracellular field potential signal of cell,
The sample frequency of signal path is the integral multiple of sinusoidal exciting signal frequency, realizes cell resistance antinoise signal and the extracellular field electricity of cell
Position the noiseless of signal detects simultaneously.
Further, in the step (1), transplantation of cardiomyocytes density is that 10-15 ten thousand is every square centimeter.
Further, the sensitive frequency that the anti-sensor of cell resistance is determined using Measured By Impedance Spectroscopy, using the sensitive frequency as
Exciting signal frequency.
Further, anti-to cell resistance to carry out continuous detection for a long time with extracellular field potential signal, to avoid experiment from tying
Fruit data volume is excessive, adjusts and at least carried out the anti-sampling with extracellular field potential signal of cell resistance every 10 minutes, samples simultaneously
Time adjustment is at least 10 seconds.
It is examined simultaneously the invention has the benefit that resisting the present invention provides a kind of cell resistance with extracellular field potential high throughput
It is noiseless simultaneously to realize two kinds of signals by sample frequency selection and notch filter double shield for examining system and method
Detection, and have the characteristics that high sensitivity, flux are high.The present invention overcomes the detections of existing cell impedance single hole scan-type, special
It is not deficiency in the case where high throughput with time delay.Operation of the present invention step is simple, has continuous for a long time to cell
The advantages of detection, applicable object area are extensive.According to the above advantage, system and method for the invention can be used for biosensor
The especially related fields of cell sensor detection.
Detailed description of the invention
Fig. 1 is that cell resistance of the present invention resists and the extracellular field potential high throughput block diagram of detection system realization simultaneously；
Fig. 2 is the anti-detection system structure simultaneously with extracellular field potential high throughput of the cell resistance of one embodiment of the invention
Figure；
Fig. 3 is that cell resistance resists and extracellular field potential compound sensor structure chart；
The extracellular field potential plane electrode 10kHz sample rate of Fig. 4 cell detects verifying to the emulation of 10kHz pumping signal；
Actually detected verifying of the extracellular field potential plane electrode 10kHz sample rate of Fig. 5 cell to 10kHz pumping signal；
In figure: pedestal 1, top cover 2, the first locating button 3, the second locating button 4, printed circuit board 5, DB44 data line interface are female
Seat the 6, the first 2.54mm row needle interface base 7, the 2nd 2.54mm row needle interface base 8, the first 2.54mm arrange needle interface 9, second
2.54mm arranges needle interface 10, electrical impedance interdigital electrode and extracellular field potential plane electrode circuit board 11, porous cell culture cavity
12, cell culture cavity protection cap 13, DC-DC power source management module 14, NI data collecting card 15, power interface 16, cell training
Support chamber 17.
Specific embodiment
Below in conjunction with attached drawing and specific example, the present invention is described in detail, but is not intended to limit the present invention.
Embodiment 1
A kind of anti-detection system simultaneously with extracellular field potential high throughput of cell resistance provided in this embodiment, as shown in Figure 1,
It include: that cell resistance is anti-electric with extracellular field potential compound sensor, the anti-conditioning circuit of cell resistance, the extracellular field potential conditioning of cell
Road and data collecting card.
Wherein, cell resistance is anti-and extracellular field potential compound sensor is by porous cell culture cavity, the anti-biography of cell resistance
Sensor and the extracellular field potential sensor composition of cell, the anti-sensor of cell resistance are made of interdigital electrode, the extracellular field potential of cell
Sensor is made of plane electrode, and two kinds of sensor combinations are at independent compound sensor unit；Porous cell culture cavity is solid
Fixed to form independent multiple cell culture chambers on the glass substrate, the bottom of each cell culture chamber is equipped with independent multiple
Close sensor unit.
Data collecting card includes excitation generation module and signal acquisition module, and excitation generation module passes through excitation conditioning circuit
Respectively with the excitation end of the anti-sensor interdigital electrode of cell resistance in the compound sensor unit of each cell culture chamber bottom
Electrical connection, for providing ac-excited signal；The test side of interdigital electrode receives signal and passes through the anti-conditioning circuit of cell resistance
Amplify, filter after be input to the signal acquisition module of data collecting card；The extracellular field potential sensor of cell passes through cell born of the same parents
Outer field potential conditioning circuit amplifies, filter after be input to the signal acquisition module of data collecting card.
Further, which further includes power management module, for being the anti-conditioning circuit of cell resistance and cell
Extracellular field potential conditioning circuit power supply.
Further, each independent compound sensor unit includes two pairs of interdigital electrodes and two plane electrodes；Two pairs
Interdigital electrode symmetry arrangement, excitation end share, and test side is separated, exports the anti-detection signal of two-way cell resistance；Two plane electricity
Pole exports the extracellular field potential detection signal of cell all the way respectively.
Further, excitation conditioning circuit will motivate the ac-excited signal of generation module output to decay to to be checked first
The undamaged voltage range of the cell of survey, is then filtered, and filters out the excitation end that interdigital electrode is input to after noise jamming.
Further, the anti-conditioning circuit of cell resistance includes current-voltage conversion circuit, high-pass filter and voltage follow
Device, current-voltage conversion circuit are used to convert the current signal that interdigital electrode test side detects to voltage signal, high pass filter
Wave device is used for influence of the isolation signals output end to input terminal for removing baseline drift, voltage follower.
Further, the extracellular field potential conditioning circuit of cell include low noise pre-amplification circuit, trap-high-pass filter,
Two-stage amplifier and voltage follower, low noise pre-amplification circuit are used for the extracellular field potential of cell that enlarged plan electrode detection arrives
Signal is detected, trap-high-pass filter is for filtering out pumping signal and baseline drift, and two-stage amplifier is for being further amplified inspection
It surveys signal and is used for influence of the isolation signals output end to input terminal to grade, voltage follower is lied prostrate.
A kind of cell resistance is anti-with extracellular field potential high throughput Simultaneous Detection, comprising the following steps:
(1) myocardial cells culture liquid is prepared, by a certain number of transplantation of cardiomyocytes in each cell culture chamber；
(2) by data collecting card generate sinusoidal excitation signal, by motivate conditioning circuit decay to cardiac muscle cell without
The voltage range of damage is applied to the excitation end of the anti-sensor interdigital electrode of cell resistance after filtering out noise, then passes through interdigital electricity
Pole test side receives current signal, since the interdigital electrode excitation end of each independent cell culture chamber shares, test side point
It opens, forms the independent multiple measure loops in every hole, therefore the no-delay cell resistance of multichannel may be implemented and resist while detecting；
(3) while step (2) detect cell resistance antinoise signal, plane electrode records the extracellular field potential signal of cell,
The sample frequency of signal path is the integral multiple of sinusoidal exciting signal frequency, realizes cell resistance antinoise signal and the extracellular field electricity of cell
Position the noiseless of signal detects simultaneously.
Further, in the step (1), transplantation of cardiomyocytes density is that 10-15 ten thousand is every square centimeter.
Further, the sensitive frequency that the anti-sensor of cell resistance is determined using Measured By Impedance Spectroscopy, using the sensitive frequency as
Exciting signal frequency.
Further, anti-to cell resistance to carry out continuous detection for a long time with extracellular field potential signal, to avoid experiment from tying
Fruit data volume is excessive, adjusts and at least carried out the anti-sampling with extracellular field potential signal of cell resistance every 10 minutes, samples simultaneously
Time adjustment is at least 10 seconds.
Embodiment 2
This gives a kind of anti-specific implementations of detection system simultaneously with extracellular field potential high throughput of cell resistance
Form, comprising: cell resistance it is anti-with extracellular field potential compound sensor, printed circuit board 5, DC-DC power source management module 14 and
NI data collecting card 15.
As shown in Fig. 2, the system further includes tool housing, tool housing is by pedestal 1, top cover 2, the first locating button 3 and
Two locating buttons 4 composition, top cover 2 are fixed on pedestal 1 by the first locating button 3 of positioning and the second locating button 4, and it is close that inside forms one
Closed chamber room, there are DB44 data-interface base 6, the first 2.54mm of compound sensor chip to arrange 7 and of needle interface base on top cover 2 surface
2nd 2.54mm arranges needle interface base 8.
Printed circuit board 5 includes the anti-conditioning circuit of cell resistance and the extracellular field potential conditioning circuit of cell, the extracellular field of cell
Current potential conditioning circuit is provided with 10kHz notch filter circuit.DC-DC power source management module 14 is both secured to printed circuit board 5
In tool housing.
Cell resistance is anti-and extracellular field potential compound sensor includes the first 2.54mm row's needle interface 9 and the 2nd 2.54mm row
Needle interface 10, electrical impedance interdigital electrode and extracellular field potential plane electrode circuit board 11, porous cell culture cavity 12 and cell
Cavity protection cap 13 is cultivated, porous cell culture cavity 12 and cell culture cavity protection cap 13 are by polymethyl methacrylate
Material customization is process, and porous cell culture cavity 12 is (ultraviolet solid by the high light-sensitive emulsion of biocompatibility with substrate of glass
Change glue) bonding, independent multiple cell culture chambers 17 are formed, as shown in figure 3, the anti-interdigital electrode of cell resistance and extracellular field electricity
Bit plane electrode uniformly, is regularly distributed in the bottom of cell culture chamber.The cell resistance resists and the compound biography of extracellular field potential
Sensor arranges the first 2.54mm on needle interface 9 and the 2nd 2.54mm row needle interface 10, with printed circuit board 5 by the first 2.54mm
Row's needle interface base 7 is connected with the 2nd 2.54mm row's needle interface base 8, has detachable replacement function.
DC-DC power source management module 14 passes through power interface 16 and the anti-conditioning circuit of cell resistance and the extracellular field potential of cell
Conditioning circuit printed circuit board is connected, and realizes power supply.
NI data collecting card 15 passes through DB44 data connecting line and the anti-conditioning circuit of cell resistance and the extracellular field potential of cell
Conditioning circuit printed circuit board surface interface base 6 is connected, and realizes excitation generation and Signals collecting function.
Using said detecting system to the anti-method detected simultaneously with extracellular field potential of cell resistance, including following step
It is rapid:
1, this experiment uses primary characterization element of the cardiac muscle cell as cell sensor, needs to configure in advance corresponding thin
Born of the same parents' culture solution: by taking HL-1 myocardial cells culture liquid as an example, 100mL culture solution includes Claycomb Medium 87mL, tire ox blood
(Fetal bovine serum) 10mL clearly, the dual anti-solution 1mL of penicillin/streptomycin (Penicillin/Streptomycin,
Ultimate density is 100ug/mL), norepinephrine 1mL (Norepinephrine, ultimate density 0.1mM), L- glutamy
Amine 1mL (L-Glutamine, ultimate density 2mM) is sterilized after preparation using the aperture 60um strainer filtering, and bottleneck uses
Sealed membrane sealing, is kept in dark place under the conditions of being placed in 20 DEG C.
2, it takes out the culture solution deposited under 20 DEG C of environmental protections and uses 200uL specification liquid relief after 37 DEG C of heating water baths
Rifle is drawn 100uL culture solution and is respectively placed in cell culture chamber 17, the electrode surface of uniform fold culture chamber hole bottom, replacement
100uL (20w/ml cell density) HL-1 cell/culture solution mixed solution is drawn after pipette tips again to be added in cell culture chamber 17,
Gently piping and druming mixes.Cell resistance is resisted and extracellular field potential compound sensor passes through the first 2.54mm row's needle interface 9 and second
The first 2.54mm row's needle interface base 7 and the 2nd 2.54mm row's needle that 2.54mm row's needle interface 10 is inserted on printed circuit board 5 connect
In mouth base 8, the connection of cell resistance anti-and extracellular field potential compound sensor and printed circuit board is realized.
3, using DB44 data line by NI data collecting card 15 and the anti-conditioning circuit of cell resistance and the extracellular field potential of cell
The DB44 data-interface base 6 of conditioning circuit printed circuit board is connected, and realizes the object of NI data collecting card 15 and printed circuit board 5
Reason communication.Generating peak-to-peak value by LabView process control NI data collecting card 15 is 1V, and frequency is the sinusoidal excitation of 10kHz,
The excitation electrode of the anti-sensor interdigital electrode of cell resistance is applied to after overdriving conditioning circuit and decaying to peak-to-peak value as 10mV
One end, then multichannel current signal is received simultaneously by detecting electrode one end in interdigital electrode system, signal sampling frequencies are
100kHz finally calculates the anti-value of cell resistance.At the same time, plane electrode records cell born of the same parents simultaneously on cell sensor chip
Outfield electric potential signal, sample frequency 10kHz set sample rate as 10k item through Fig. 4-5 emulation and actually detected result verification
Under part, the pumping signal of 10kHz is through extracellular field potential conditioning circuit post-sampling will not (the extracellular field potential of cell be believed to useful signal
Number) generate aliazing effect.
4, to avoid data volume excessive, host computer LabView control NI data collecting card 15, which is spaced every 10 minutes, to be carried out carefully
Born of the same parents' electrical impedance and the continuous of extracellular field potential signal monitor and are saved in local file, the sample duration of two of them signal
It is 10 seconds.It is every after 24 hours in cell culture, it needs to carry out intracavitary cell culture fluid to change liquid, be arranged by LabView
15 time out of NI data collecting card is arranged from the first 2.54mm and is taken in needle interface base 7 and the 2nd 2.54mm row's needle interface base 8
Lower cell resistance resists and extracellular field potential compound sensor chip, is transferred to super-clean bench replacement fresh cell medium, more
The cell culture fluid changed still keeps the total volume of 200uL, hereafter resists cell resistance and extracellular field potential compound sensor core
Piece arranges needle interface 9 by the first 2.54mm and the 2nd 2.54mm arranges needle interface 10 and is inserted into the anti-conditioning circuit of cell resistance and cell born of the same parents
The first 2.54mm row's needle interface base 7 and the 2nd 2.54mm row's needle interface on outer field potential conditioning circuit printed circuit board 5 is female
In seat 8, the signal detection of next round is carried out.
5, experiment terminates, and stops 15 program process of NI data collecting card of LabView control, and continuous monitoring data are stored in
In " local file " that step 4 is previously mentioned, disconnect power module power supply, finally remove compound sensor chip cleaning after prepare into
Row is tested next time.
Above-mentioned specific embodiment is used to illustrate the present invention, rather than limits the invention, of the invention
In spirit and scope of protection of the claims, to any modifications and changes that the present invention makes, protection model of the invention is both fallen within
It encloses.
Claims (10)
1. a kind of anti-detection system simultaneously with extracellular field potential high throughput of cell resistance, which is characterized in that anti-including cell resistance
With extracellular field potential compound sensor, the anti-conditioning circuit of cell resistance, the extracellular field potential conditioning circuit of cell and data collecting card；
Wherein, cell resistance is anti-and extracellular field potential compound sensor is by porous cell culture cavity, the anti-sensor of cell resistance
With the extracellular field potential sensor composition of cell, the anti-sensor of cell resistance is made of interdigital electrode, the extracellular field potential sensing of cell
Device is made of plane electrode, and two kinds of sensor combinations are at independent compound sensor unit；Porous cell culture cavity is fixed on
In substrate of glass, independent multiple cell culture chambers are formed, the bottom of each cell culture chamber is equipped with independent compound biography
Sensor cell；
Data collecting card includes excitation generation module and signal acquisition module, and excitation generation module passes through excitation conditioning circuit difference
It is electrically connected with the excitation end of the anti-sensor interdigital electrode of cell resistance in the compound sensor unit of each cell culture chamber bottom
It connects, for providing ac-excited signal；The test side of interdigital electrode receives signal and is carried out by the anti-conditioning circuit of cell resistance
The signal acquisition module of data collecting card is input to after amplification, filtering；The extracellular field potential sensor of cell passes through the extracellular field of cell
Current potential conditioning circuit amplifies, filter after be input to the signal acquisition module of data collecting card.
2. a kind of anti-detection system simultaneously with extracellular field potential high throughput of cell resistance according to claim 1, feature
It is, which further includes power management module, for being the anti-conditioning circuit of cell resistance and the extracellular field potential tune of cell
Manage circuit power supply.
3. a kind of anti-detection system simultaneously with extracellular field potential high throughput of cell resistance according to claim 1, feature
It is, each independent compound sensor unit includes two pairs of interdigital electrodes and two plane electrodes；Two pairs of interdigital electrodes are symmetrical
Arrangement, excitation end share, and test side is separated, exports the anti-detection signal of two-way cell resistance；Two plane electrodes export one respectively
Cell extracellular field potential in road detects signal.
4. a kind of anti-detection system simultaneously with extracellular field potential high throughput of cell resistance according to claim 1, feature
It is, the excitation conditioning circuit will motivate the ac-excited signal of generation module output to decay to cell to be detected first
Undamaged voltage range, is then filtered, and filters out the excitation end that interdigital electrode is input to after noise jamming.
5. a kind of anti-detection system simultaneously with extracellular field potential high throughput of cell resistance according to claim 1, feature
It is, the anti-conditioning circuit of cell resistance includes current-voltage conversion circuit, high-pass filter and voltage follower, electric current-
Voltage conversion circuit is used to convert voltage signal for the current signal that interdigital electrode test side detects, high-pass filter is used for
Baseline drift is removed, voltage follower is used for influence of the isolation signals output end to input terminal.
6. a kind of anti-detection system simultaneously with extracellular field potential high throughput of cell resistance according to claim 1, feature
It is, the extracellular field potential conditioning circuit of cell includes low noise pre-amplification circuit, trap-high-pass filter, second level amplification
Device and voltage follower, low noise pre-amplification circuit is for the extracellular field potential detection letter of cell that enlarged plan electrode detection arrives
Number, trap-high-pass filter is for filtering out pumping signal and baseline drift, and two-stage amplifier is for being further amplified detection signal
To volt grade, voltage follower is used for influence of the isolation signals output end to input terminal.
7. a kind of examine using any one of the claim 1-6 system progress cell resistance is anti-with extracellular field potential high throughput simultaneously
The method of survey, which comprises the following steps:
(1) myocardial cells culture liquid is prepared, by a certain number of transplantation of cardiomyocytes in each cell culture chamber；
(2) sinusoidal excitation signal is generated by data collecting card, it is not damaged to cardiac muscle cell by motivating conditioning circuit to decay to
Voltage range, filter out the excitation end that the anti-sensor interdigital electrode of cell resistance is applied to after noise, then pass through interdigital electrode and examine
It surveys end and receives current signal, since the interdigital electrode excitation end of each independent cell culture chamber shares, test side is separated, shape
At the independent multiple measure loops in every hole, therefore the no-delay cell resistance of multichannel may be implemented and resist while detecting；
(3) while step (2) detect cell resistance antinoise signal, plane electrode records the extracellular field potential signal of cell, signal
The sample frequency in channel is the integral multiple of sinusoidal exciting signal frequency, realizes cell resistance antinoise signal and the extracellular field potential letter of cell
Number noiseless detect simultaneously.
8. the method according to the description of claim 7 is characterized in that transplantation of cardiomyocytes density is 10- in the step (1)
150000 is every square centimeter.
9. the method according to the description of claim 7 is characterized in that determining the quick of the anti-sensor of cell resistance using Measured By Impedance Spectroscopy
Frequency is felt, using the sensitive frequency as exciting signal frequency.
10. being grown the method according to the description of claim 7 is characterized in that anti-to cell resistance with extracellular field potential signal
Time Continuous detection, to avoid experimental result data amount excessive, adjust at least every 10 minutes progresss cell resistances resist with it is extracellular
The sampling of field potential signal, while the sampling time is adjusted at least 10 seconds.
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| CN113528332A (en) * | 2021-07-19 | 2021-10-22 | 中山大学 | Intracellular and extracellular electrophysiological recording system and method for automatic electroporation regulation and screening |
| CN114292749A (en) * | 2022-01-19 | 2022-04-08 | 中山大学 | Device and method for synchronously detecting high-throughput intracellular and extracellular electrophysiological and mechanical pulsation activities of electrically excited cells |
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| CN111239197A (en) * | 2020-02-21 | 2020-06-05 | 中山大学 | Device and method for integrating cell full-spectrum impedance and rapid impedance measurement |
| CN113528332A (en) * | 2021-07-19 | 2021-10-22 | 中山大学 | Intracellular and extracellular electrophysiological recording system and method for automatic electroporation regulation and screening |
| CN113528332B (en) * | 2021-07-19 | 2023-10-24 | 中山大学 | Intracellular and extracellular electrophysiological recording system and method for automatic electroporation regulation screening |
| CN114292749A (en) * | 2022-01-19 | 2022-04-08 | 中山大学 | Device and method for synchronously detecting high-throughput intracellular and extracellular electrophysiological and mechanical pulsation activities of electrically excited cells |
| CN114292749B (en) * | 2022-01-19 | 2023-10-24 | 中山大学 | Device and method for synchronously detecting high-flux intracellular and extracellular electrophysiological and mechanical pulsation activity of electrically excited cells |
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