CN111289819B - Integrated recording regulation and control system for measuring intracellular electric signals by myocardial cell electroporation - Google Patents
Integrated recording regulation and control system for measuring intracellular electric signals by myocardial cell electroporation Download PDFInfo
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Abstract
The invention discloses an integrated recording regulation and control system for measuring intracellular electric signals by myocardial cell electroporation, which comprises an upper computer, a PCB substrate, a hollow platinum nanotube array sensor fixed on the PCB substrate, a sensor electric signal conditioning circuit, an electroporation signal output line and a signal acquisition card. The hollow platinum nanotube array sensor comprises a PCB (printed Circuit Board), a working electrode and a reference electrode, wherein the working electrode is connected with the input end of an electric signal conditioning circuit of the sensor; the reference electrode is grounded. A cylinder body is fixed on the working electrode and the reference electrode and is used as a cell culture cavity. The microelectrode consists of a PET film and a hollow platinum nanotube array which grows on the PET film and has the diameter of 0.4-1 μm and the length of 0.5-2 μm. The system of the invention adopts a minimally invasive electroporation method to record the intracellular electric signals of the myocardial cells, simultaneously ensures that the basic activity of the myocardial cells is not influenced, and adopts a non-marking signal recording method to automatically record the intracellular electric signals of the myocardial cells for a long time.
Description
Technical Field
The invention relates to an automatic myocardial cell intracellular electric signal recording device, in particular to an integrated recording regulation and control system capable of carrying out myocardial electroporation and intracellular electric signals.
Background
Since myocardial cells are the basic unit of the heart and can generate action potentials rhythmically to generate mechanical contraction, which are the basic biological functions of the heart to pump blood, the study of myocardial cell electrophysiology plays an important role in the field of cardiology. At present, the patch clamp technology is a commonly used method for recording an intracellular action potential signal of a myocardial cell, although the technology can effectively record a standard action potential signal, the invasive detection principle of the technology has great damage to the myocardial cell, and long-term intracellular electrophysiological research of the myocardial cell cannot be carried out. The recording of the extracellular electrical signals of the cardiomyocytes based on the microelectrode array can non-invasively record the extracellular electrical signals for a long time, and the signals can reflect the electrophysiological state of the cardiomyocytes to a certain extent.
Disclosure of Invention
Aiming at the problem that the intracellular electric signals of the myocardial cells cannot be recorded stably for a long time in the prior art, the invention develops a myocardial electroporation and intracellular electric signal integrated recording and regulating system based on a conductive nano needle tube array, and automatically performs long-time recording on the intracellular electric signals of the myocardial cells by adopting a minimally invasive electroporation method and a non-labeled signal recording method.
The purpose of the invention is realized by the following technical scheme:
an integrated recording regulation and control system for measuring intracellular electric signals by myocardial cell electroporation comprises an upper computer, a PCB substrate, a hollow platinum nanotube array sensor fixed on the PCB substrate, a sensor electric signal conditioning circuit, an electroporation signal output line and a signal acquisition card.
The hollow platinum nanotube array sensor comprises a PCB (printed Circuit Board), a working electrode and a reference electrode, wherein the electrode end of the working electrode is 6.5mm long and 20-100 mu m wide, and the connecting end is 5mm long and 2mm wide; the electrode end of the reference electrode is 4.5mm long and 20-100 μm wide, and the connecting end is 8mm long and 2mm wide. The working electrode is connected with the input end of the sensor electric signal conditioning circuit; the reference electrode is grounded. The working electrode and the reference electrode are fixed on the PCB through the connecting end. A cylinder body is fixed on the working electrode and the reference electrode and is used as a cell culture cavity.
The working electrode and the reference electrode are both composed of a PET film and a hollow platinum nanotube array with the diameter of 0.4-1 μm and the length of 0.5-2 μm, which grows on the PET film.
The upper computer is used for controlling the electroporation signal output, displaying and recording data obtained by the hollow platinum nanotube array sensor test.
The sensor electric signal conditioning circuit consists of a first-stage amplifying circuit, a low-pass filter, an RC DC blocking circuit and a second-stage amplifying circuit which are connected in sequence, wherein the first-stage amplifying circuit and the second-stage amplifying circuit are both in-phase proportional amplifiers; the RC direct current blocking circuit is composed of a 10 muF capacitor C7, a resistor R16 and a resistor R17, wherein the resistance values of the resistor R16 and the resistor R17 are both 30k omega, and the resistor R16 and the resistor R17 are connected in series and then are connected in parallel with a capacitor C7; two ends of the capacitor C7 are respectively connected with the output end of the low-pass filter and the input end of the second-stage amplifying circuit; the output end of the second-stage amplifying circuit is connected with a signal acquisition card, and the signal output end of the signal acquisition card is connected with the input end of the upper computer. The electroporation signal output line is connected with the analog output module of the signal acquisition card.
Furthermore, the number of the working electrodes and the number of the reference electrodes are multiple, the working electrodes and the reference electrodes are distributed in a central symmetry manner, and the distances between all the working electrodes and the reference electrodes are not more than 1 mm.
Furthermore, the hollow platinum nanotube array sensor is provided with a pin header, the PCB substrate is provided with corresponding jacks, the pin header is connected with the corresponding working electrode and the reference electrode, and the jacks are connected with the sensor electrical signal conditioning circuit.
Further, the working electrode and the reference electrode are prepared by the following steps:
(1) photoetching to obtain electrode patterns of a working electrode and a reference electrode: taking a PET film with the aperture of 450nm as an insulating substrate, and preparing an electrode end of a working electrode by photoetching, wherein the electrode end is 6.5mm long, the width is 20-100 mu m, and the connecting end is 5mm long and 2mm wide; and (3) obtaining the patterned PET film by an electrode arrangement pattern with the electrode end of the reference electrode being 4.5mm long, the width being 20-100 mu m, the connecting end length being 8mm and the width being 2 mm.
(2) And (3) plating an electrode, namely performing magnetron sputtering of 30nm gold or platinum on the patterned PET film obtained in the step (1), and removing the gold or platinum outside the electrode arrangement pattern to obtain the conductive PET film.
(3) Preparing a hollow platinum nanotube array: using a metal-plated surface contact copper sheet of a PET film as a working electrode, an Ag/AgCl electrode as a reference electrode, a platinum wire as a counter electrode, and performing electrodeposition for 200s in a constant current working mode by using an electrolyte containing 1 wt% of chloroplatinic acid and 0.5M hydrochloric acid on a conductive PET filmThe hole wall forms a hollow platinum nano-tubular structure. Reuse of O2And etching away part of PET on the non-sputtered metal surface on the PET film by the plasma, and exposing the electrode arrangement hollow platinum nanotube array with the diameter of 0.4-1 μm and the length of 0.5-2 μm.
Furthermore, the first-stage amplifying circuit further comprises a capacitor, and the capacitor is arranged between the reverse input end and the output end of the operational amplifier in the in-phase proportional amplifier. The capacitor can be used for lag compensation and preventing the self-oscillation of the in-phase proportional amplifier. The low-pass filter is a Sallen-Key low-pass filter consisting of a low-noise operational amplifier U2A, a capacitor C5 of 1 nF-100 nF, a capacitor C6, a resistor R12 of which the resistance value is 1 k-50 k, a resistor R13, a resistor R15, a resistor R20 and a resistor R21.
The method has the advantages that the method can record the intracellular electric signals of the myocardial cells and simultaneously ensure that the basic activity of the myocardial cells is not influenced, thereby realizing the automatic long-term recording of the intracellular electric signals of the myocardial cells.
Drawings
FIG. 1 is a scanning electron microscope characterization of hollow platinum nanotubes;
FIG. 2 is a schematic diagram of a hollow platinum nanotube array sensor;
FIG. 3 is a schematic diagram of a hollow platinum nanotube array for recording and modulating cardiomyocytes;
FIG. 4 is a block diagram of a sensor electrical signal conditioning and electroporation circuit;
FIG. 5 is a block diagram of a myocardial electroporation and intracellular electrical signal integrated recording and control system for hollow platinum nanotube arrays;
FIG. 6 is a schematic diagram of a sensor electrical signal conditioning and electroporation circuit;
FIG. 7 is a program flow diagram of the upper computer control software;
FIG. 8 is a main interface of an upper computer of the myocardial electroporation and intracellular electrical signal integrated recording regulation and control system of the hollow platinum nanotube array;
FIG. 9 is a diagram of the results of real-time data acquisition of the myocardial electroporation and intracellular electrical signal integrated recording regulation and control system of hollow platinum nanotube arrays.
In the figure, a working electrode 1, a reference electrode 2, a PCB 3, a pin header 4, a cell culture cavity 5, a PET film 6, a hollow platinum nanotube array 7, a cell 8, a pin header jack 9, a hollow platinum nanotube array sensor 10, a sensor electrical signal conditioning circuit 11, a conditioned sensor electrical signal output end 12, a power supply interface 13, a conditioned sensor electrical signal output end 14, a PCB substrate 15, an acquisition card No. 1 16, an acquisition card No. 2 17 and an electroporation signal output line 18.
Detailed Description
The detection principle of the intracellular electrical signal of the cardiomyocytes used is described in detail below.
The myocardial cell is the basic component cell of the heart, has electrical excitability, and can generate action potential signals autonomously, the action potential generation is caused by the inflow and outflow of corresponding ion channels of sodium ions, potassium ions and calcium ions on a cell membrane, the formed action potential can be conducted to the outside of the cell through the cell membrane of the myocardial cell, so that extracellular field signals of the myocardial cell are formed, and the extracellular signals can be recorded by a common microelectrode array. In order to record the intracellular electric signals, a nano electrode array electrode is adopted, electroporation is carried out through an analog output module of a data acquisition card, so that the tip of the hollow platinum nanotube array of the microelectrode 1 on the sensor 10 discharges, micro nano cracks are generated on cell membranes, the nano electrode array can record the intracellular electric signals, and the intracellular electric signals are amplified and filtered by a high-input-impedance low-noise amplifier, so that the intracellular electric signals are acquired and analyzed by the analog input module of the data acquisition card.
The purpose and effect of the present invention will be better demonstrated by further explaining the present invention with reference to the examples and the accompanying drawings.
As shown in FIG. 4, the integrated recording and regulating system for measuring the intracellular electrical signals by the electroporation of the cardiomyocytes comprises an upper computer, a PCB substrate 15, a hollow platinum nanotube array sensor 10 fixed on the PCB substrate 15, a sensor electrical signal conditioning circuit 11, an electroporation signal output line 18 and a signal acquisition card.
As shown in fig. 2, the hollow platinum nanotube array sensor 10 includes a PCB 3, a working electrode 1 and a reference electrode 2, wherein the electrode end of the working electrode 1 is 6.5mm long and 20-100 μm wide, and the connecting end is 5mm long and 2mm wide; the electrode end of the reference electrode 2 is 4.5mm long and 20-100 μm wide, and the connection end is 8mm long and 2mm wide. The working electrode 1 is connected with the input end of the sensor electric signal conditioning circuit 11; the reference electrode 2 is grounded. The working electrode 1 and the reference electrode 2 are fixed on the PCB 3 through connecting ends, and the connecting ends are connected with bonding pads on the PCB 3 through conductive silver paste. A cylinder body is fixed on the working electrode 1 and the reference electrode 2 to be used as a cell culture cavity 5.
Preferably, the hollow platinum nanotube array sensor 10 has a plurality of working electrodes 1 and reference electrodes 2, and the working electrodes 1 and the reference electrodes 2 are distributed in central symmetry so as to be conveniently bonded with the cylindrical cell culture chamber 5. The distances between all the working electrodes 1 and the reference electrode 2 are not more than 1mm, so that the cell impedance is too large, and the circuit cannot detect signals. Fig. 2 is a schematic structural diagram of a hollow platinum nanotube array sensor 10 with 16 working electrodes 1 and 4 reference electrodes 2.
The working electrode 1 and the reference electrode 2 are both composed of a PET film 6 and a hollow platinum nanotube array 7 with the diameter of 0.4-1 μm and the length of 0.5-2 μm, which is grown on the PET film 6. As shown in FIG. 3, the electrode with the three-dimensional structure has better coupling effect with the cell membrane of the cell 8 on the surface thereof, which is beneficial to improving the detection sensitivity of signals.
In this embodiment, a polyethylene terephthalate (PET) polymer film-PET film 6 having a pore diameter of 0.4 to 1 μm is used as a template to prepare a hollow platinum nanotube electrode array 7. Firstly, a layer of RZJ-390PG positive photoresist is spin-coated on a PET film 6 by adopting a photoetching technology, ultraviolet light irradiates the surface of the PET film through a mask plate for exposure, and after the PET film is soaked in a developing solution, the photoresist of the exposed part is removed, thus obtaining the patterned PET film 6. Then, magnetron sputtering 30nm gold or platinum is carried out on the developed PET film 6, and then acetone is adopted to dissolve the residual photoresist, so that the conductive PET film 6 can be obtained. Then adopting an electrochemical deposition technology, taking a metal-plated surface contact copper sheet of the PET film 6 as a working electrode, an Ag/AgCl electrode as a reference electrode and a platinum wire as a counter electrode to form a three-electrode system, and immersing the PET film 6 into electrolyte which comprises 1 wt% of chloroplatinic acid and 0.5M hydrochloric acid. Electrodepositing for 200s in constant current/constant voltage working mode on PET film6, the hole wall forms a hollow platinum nano-tubular structure. Followed by O2The upper surface of the non-sputtered metal surface of the PET film 6 is etched by plasma, and a hollow platinum nanotube array 7 structure with the diameter of 0.4-1 μm and the length of 0.5-2 μm is exposed.
The hollow platinum nanotube electrode array 7 is upward, the connecting end part and the PCB plate 3 are sequentially connected and fixed by conductive silver paste on the bottom surface, then the upper row of pins 4 are welded, finally the cell culture cavity 5 (the diameter is about 1.4cm) is bonded by uncured PDMS on the upper part of the electrode, and the cell culture cavity is placed for 2 hours at the temperature of 80 ℃ to cure the PDMS.
The signal conditioning circuit comprises a first-stage amplifying circuit, an RC (resistor-capacitor) DC blocking circuit, a low-pass filter and a second-stage amplifying circuit, as shown in fig. 6, the first-stage amplifying circuit comprises a first operational amplifier U1A, a capacitor C8 arranged between the reverse input end and the output end of the first operational amplifier U1A, a resistor R19 connected with the capacitor C8 in parallel, a resistor R23 with one end connected with the reverse input end of the first operational amplifier U1A, and a resistor R11 with one end connected with the forward input end of the first operational amplifier U1A, wherein the other end of the resistor R11 is connected with a working electrode, the other end of the resistor R23 is connected with the ground, and the reference electrode 2 is connected with the ground. The resistances of resistors R11, R19 and R23 are all between 1k omega-200 k omega, the capacitance value of a feedback capacitor C8 is between 22 pF-1 nF, the function of the feedback capacitor C8 is phase compensation, self-oscillation of U1 8 is prevented, the low-pass filter is a Sallen-8 structure low-pass filter formed by a low-noise operational amplifier U2 8, a capacitor C8 of 1 nF-100 nF, a resistor R8 and a resistor R8 with the resistance value of 1 k-50 k, a resistor R8 and a resistor R8, the forward input end of the low-noise operational amplifier U2 8 is connected with the capacitor C8 and the resistor R8, the other end of the C8 is grounded, the other end of the resistor R8 is respectively connected with the resistor R8 and the capacitor C8, the R8 is connected with the output ends of the resistors R8 and the U1 8, the other end of the R8 is connected with the output end of the U2 8, the U8 and the reverse phase feedback circuit is connected with the U8; the RC direct-current isolating circuit consists of a 10 muF capacitor C7, a 30K omega resistor R16 and a resistor R17, the resistor R16 and the resistor R17 are connected in series and then connected in parallel with the capacitor C7, and one end of the capacitor C7 is connected with the output end of the second operational amplifier U2A; the second-stage amplifying circuit consists of a precision operational amplifier U3A, a resistor R5 with one end connected with the output end of the precision operational amplifier U3A, a resistor R14 of 1 k-49.9 k, a resistor R18 and a resistor R22. The resistor R18 is arranged between the reverse input end and the output end of the precision operational amplifier U3A, one end of the resistor R22 is connected with the reverse input end of the precision operational amplifier U3A, and the other end of the resistor R22 is grounded; one end of the resistor R14 is connected with the positive input end of the precision operational amplifier U3A, and the other end is connected with the capacitor C7. One end of the resistor R5 is grounded, and the other end of the resistor R5 is connected to the output end of the precision operational amplifier U3A; the output end of the precision operational amplifier U3A is connected with a signal acquisition card, the signal output end of the signal acquisition card is connected with the input end of an upper computer, and the other channel of the same signal conditioning circuit is the same as the circuit described above. The electroporation signal output line 18 is connected to the analog output channel of the signal acquisition card.
Electronic components related to the circuit are welded on the sensor electrical signal conditioning circuit module, and each path has two channels by adopting double operational amplifiers, then another 7 paths of identical sensor electrical signal conditioning circuits 11 are needed on the board to form a 16-channel sensor electrical signal conditioning circuit 11, which corresponds to 16 working electrodes in fig. 2, the eight-channel signal conditioning circuit on the right side of the sensor electrical signal conditioning and electroporation circuit module is connected with the signal output end 12, the four-way eight-channel signal conditioning circuit of the sensor electrical signal conditioning and electroporation circuit module is connected with the signal output end 13, the power supply interface 13 is connected with external +/-5V voltage, then the signal output end 12 is connected with the data acquisition card 16 by a silica gel flat cable, the signal output end 14 is connected with the data acquisition 17 by a silica gel flat cable, and the data acquisition card 16 and the data acquisition card 17 are connected with an upper computer after being connected with the USB concentrator. The upper computer is used for controlling the electroporation signal output, displaying and recording the data obtained by the test of the hollow platinum nanotube array sensor 10.
Preferably, the working electrode 1 and the reference electrode 2 are structurally connected with the sensor electrical signal conditioning circuit 11 through the pin header 4 and the jack 9, the pin header 4 is arranged on the hollow platinum nanotube array sensor 10, the corresponding jack 9 is arranged on the PCB substrate 15, the pin header 4 is connected with the corresponding working electrode 1 and the reference electrode 2, and the jack 9 is connected with the sensor electrical signal conditioning circuit 11.
As shown in fig. 5, the working process of the present invention is as follows: the method comprises the steps of enabling one side of a hollow platinum nanotube array 7 to face upwards, inserting a pin header 4 of a hollow platinum nanotube array sensor 10 into a corresponding pin header slot 9, culturing mouse cardiac muscle cells 8 on the pin header, inserting an electroporation signal output line 18 into a culture cavity 5, starting a +/-5V power supply, enabling a sensor electrical signal conditioning circuit 18 to work, clicking an upper computer to start collecting, controlling a data acquisition card to output an electroporation pulse signal by a PC (personal computer), enabling electroporation time to be less than 100ms, enabling electroporation voltage to be generally less than 10mV, enabling the mouse cardiac muscle cells in the culture cavity to generate intracellular electrical signals, transmitting the intracellular electrical signals to an input end of the sensor electrical signal conditioning circuit 11 through the connected pin header 4, amplifying the electrical signals through an in-phase proportional amplifier firstly, then reducing high-frequency noise through low-pass filtering, filtering a baseline through an RC (resistor) DC (direct current) isolating circuit, amplifying for the second stage, and finally transmitting the electrical signals to the data acquisition card 16, And 17, transmitting the collected data to an upper computer for displaying and recording.
Preferably, as shown in fig. 7-8, the upper computer of the regulating and controlling system for recording the myocardial electroporation and intracellular electrical signals of the hollow platinum nanotube array has the following working procedures: opening a program entry interface, selecting whether to record data or not, clicking to start acquisition, recording the start time of an experiment according to the current system time by an upper computer, simultaneously establishing a TDMS recording file under the current directory by taking the start time as a naming format, informing the acquisition card to send a pulse voltage signal by the upper computer, and simultaneously informing the acquisition card to acquire data, sending the acquired data into a buffer queue by one thread of the upper computer, extracting the data of the buffer queue according to a first-in first-out rule by the other thread, reducing the data by an amplification factor, and sending the data to a waveform chart for displaying and recording the TDMS file. When a certain amount of data is collected, the user may click the start collection button again, at which point the button transitions to a stop indicating that it is currently in a stopped state.
The following gives examples of applications of the present invention.
The myocardial electroporation and intracellular electric signal integrated recording regulation and control system of the hollow platinum nanotube array is mainly used for detecting myocardial intracellular electric signals. In the experiment, firstly, the myocardial cells and the culture solution thereof are added into a culture cavity 5, a hollow platinum nanotube array sensor 10 is inserted into a slot 9, a sensor electrical signal conditioning circuit 11, a data acquisition card, a USB concentrator and an upper computer are connected by using a silica gel flat cable and data, an electroporation signal output line 18 is inserted into the culture cavity 5, a +/-5V power supply switch is turned on, upper computer software is turned on, a main interface is accessed as shown in figure 8, data is selected and recorded, the data is clicked and turned on, and after waiting for about 1 second, the data is displayed on a waveform chart as shown in figure 9, wherein the graph contains 16-channel data.
The invention adopts the conductive metal nano needle tube array, develops and applies an automatic electroporation and intracellular electric signal recording system, and realizes the safe and stable long-term recording of the myocardial cell intracellular electric signals.
Claims (5)
1. An integrated recording regulation and control system for measuring intracellular electric signals by myocardial cell electroporation is characterized by comprising an upper computer, a PCB substrate (15), a hollow platinum nanotube array sensor (10) fixed on the PCB substrate (15), a sensor electric signal conditioning circuit (11), an electroporation signal output line (18) and a signal acquisition card;
the hollow platinum nanotube array sensor (10) comprises a PCB (printed circuit board) (3), a working electrode (1) and a reference electrode (2), wherein the electrode end of the working electrode (1) is 6.5mm long and 20-100 mu m wide, and the connecting end is 5mm long and 2mm wide; the electrode end of the reference electrode (2) is 4.5mm long and 20-100 μm wide, and the connecting end is 8mm long and 2mm wide; the working electrode (1) is connected with the input end of the sensor electric signal conditioning circuit (11); the reference electrode (2) is grounded; the working electrode (1) and the reference electrode (2) are fixed on the PCB (3) through the connecting end; a cylinder body is fixed on the working electrode (1) and the reference electrode (2) and is used as a cell culture cavity (5);
the working electrode (1) and the reference electrode (2) are both composed of a PET film (6) and a hollow platinum nanotube array (7) which grows on the PET film (6) and has the diameter of 0.4-1 mu m and the length of 0.5-2 mu m;
the upper computer is used for controlling the electroporation signal output, displaying and recording data obtained by the test of the hollow platinum nanotube array sensor (10);
the sensor electric signal conditioning circuit (11) consists of a first-stage amplifying circuit, a low-pass filter, an RC DC blocking circuit and a second-stage amplifying circuit which are connected in sequence, wherein the first-stage amplifying circuit and the second-stage amplifying circuit are both in-phase proportional amplifiers; the RC direct current blocking circuit is composed of a 10 muF capacitor C7, a resistor R16 and a resistor R17, wherein the resistance values of the resistor R16 and the resistor R17 are both 30k omega, and the resistor R16 and the resistor R17 are connected in series and then are connected in parallel with a capacitor C7; two ends of the capacitor C7 are respectively connected with the output end of the low-pass filter and the input end of the second-stage amplifying circuit; the output end of the second-stage amplification circuit is connected with a signal acquisition card, and the signal output end of the signal acquisition card is connected with the input end of an upper computer; an electroporation signal output line (18) is connected with an analog output module of the signal acquisition card.
2. The system for integrally recording and regulating the intracellular electrical signals measured by the myocardial cell electroporation as claimed in claim 1, wherein the number of the working electrodes (1) and the reference electrodes (2) is multiple, the working electrodes (1) and the reference electrodes (2) are distributed in a centrosymmetric manner, and the distances between all the working electrodes (1) and the reference electrodes (2) are not more than 1 mm.
3. The integrated recording and regulating system for measuring the intracellular electric signals through the myocardial cell electroporation as claimed in claim 1, wherein the hollow platinum nanotube array sensor (10) is provided with a pin header (4), the PCB substrate (15) is provided with corresponding jacks (9), the pin header (4) is connected with the corresponding working electrode (1) and the reference electrode (2), and the jacks (9) are connected with the sensor electric signal conditioning circuit (11).
4. The system for integrally recording and regulating the intracellular electric signals measured by the myocardial cell electroporation as claimed in claim 1, wherein the working electrode (1) and the reference electrode (2) are prepared by the following steps:
(1) photoetching to obtain patterns of a working electrode (1) and a reference electrode (2): taking a PET film with the aperture of 450nm as an insulating substrate, and preparing an electrode end of a working electrode (1) by photoetching, wherein the electrode end is 6.5mm long, the width is 20-100 mu m, and the length and the width of the connecting end are 5mm and 2mm respectively; an electrode arrangement pattern with the electrode end of the reference electrode (2) being 4.5mm long and 20-100 μm wide, the connecting end being 8mm long and 2mm wide is obtained;
(2) plating an electrode: performing magnetron sputtering of 30nm gold or platinum on the patterned PET film obtained in the step (1), and removing the gold or platinum outside the electrode arrangement pattern to obtain a conductive PET film;
(3) preparing a hollow platinum nanotube array: taking a metal-plated surface contact copper sheet of the PET film as a working electrode, an Ag/AgCl electrode as a reference electrode, a platinum wire as a counter electrode, and electrodepositing for 200s in a constant current working mode by using an electrolyte containing 1 wt% of chloroplatinic acid and 0.5M hydrochloric acid to form a hollow platinum nano tubular structure on the hole wall of the conductive PET film; reuse of O2And etching away part of PET on the non-sputtered metal surface on the PET film by the plasma, and exposing the electrode arrangement hollow platinum nanotube array with the diameter of 0.4-1 μm and the length of 0.5-2 μm.
5. The integrated recording and control system for measuring the intracellular electric signals through the myocardial cell electroporation as claimed in claim 1, wherein the first-stage amplifying circuit further comprises a capacitor, and the capacitor is arranged between the reverse input end and the output end of the operational amplifier in the in-phase proportional amplifier; the low-pass filter is a Sallen-Key low-pass filter consisting of a low-noise operational amplifier U2A, a capacitor C5 of 1 nF-100 nF, a capacitor C6, a resistor R12 of which the resistance value is 1 k-50 k, a resistor R13, a resistor R15, a resistor R20 and a resistor R21.
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| 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 |
| CN113466110A (en) * | 2021-07-19 | 2021-10-01 | 中山大学 | Cell electrophysiological signal discrete in-situ long-time measurement and recording system and method |
| 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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