CN107462511A - Pass through the device of nano-electrode An arrayed recording endocellular electricity signal - Google Patents
Pass through the device of nano-electrode An arrayed recording endocellular electricity signal Download PDFInfo
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- CN107462511A CN107462511A CN201710570314.9A CN201710570314A CN107462511A CN 107462511 A CN107462511 A CN 107462511A CN 201710570314 A CN201710570314 A CN 201710570314A CN 107462511 A CN107462511 A CN 107462511A
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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
- 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
- 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 belongs to record the technical field of endocellular electricity signal, a kind of device by nano-electrode An arrayed recording endocellular electricity signal is specifically disclosed, including silicon materials substrate and the nano-electrode array being placed on silicon materials substrate, the position of nano-electrode array is defined using photoetching and reactive ion etching on silicon materials substrate, the position of non-nano electrod-array is covered by insulating properties silica on silicon materials substrate, the nano-electrode array is the nano-electrode of some settings, and the nano-electrode array is electrically connected between electronic component with for stimulating/measuring.The device simply and easily can enter horizontal electrical signal record to multiple cells interior at the same time, in addition, it can record extracellular electric signal and endocellular electricity signal according to being actually needed, and in addition to recording the electric signal of nerve cell, it can be also used for recording the electric signal of cardiac muscle cell or other kinds of cell.
Description
Technical field
It is more particularly to a kind of to pass through nano-electrode An arrayed recording the invention belongs to record the technical field of endocellular electricity signal
The device of endocellular electricity signal.
Background technology
In the prior art, study cerebral neuron function and connectedness all the time by huge concern, but still deposit
In many technical difficulties.In these years, with genetic marker, immunohistochemical staining, optics and photoelectric technology, electrophysiology and
The rapid development of computer technology, biomedical sector have been able to identify the type of neuron, explain their molecular mechanism,
The wiring between them is untied, the principle of neural coding is decoded and acts on the functional attributes of specific brain region.
Brain is a complicated system, it is necessary to which a set of comprehensive technology handles and understands the nerve association inside brain
Make.Endocellular electricity signal record for research occur neuron current potential generate and change electrical activity be it is essential, because
This it be to illustrate nerve cell synaptic input to stimulate the important method for the mechanism for being converted into output potential change.Pass through endocellular electricity
The record of signal, it will be appreciated that the activity of nerve cell, and cooperation and communication between nerve cell.Intracellular signals record skill
Art, which is produced and changed to nerve cell action potential, provides important research meanses, but minimally invasive, steady due to establishing in vivo
Fixed endocellular electricity signal recording system has certain difficulty, cause at present still can not effectively measuring animal brain it is potential
The electroneurographic signal subthreshold value activity change of perception, memory and behavior.Intracellular electrode passage would generally draw when recording cell
Play biochemical and physiological disturbance or even damage the activity of nerve cell.For example, using traditional Patch-clamp techniques nerve
During endocellular electricity signal, patch-clamp liquid pipe internal solution and cytoplasm fast exchange solute.This allows with inside patch-clamp liquid pipe
Solution is by the concentration of experiment control intracellular ion He other molecules, and the change of the intracellular point position of record, but this can
Biochemical process necessary to normal cell function is caused to be destroyed.Intracellular important molecule or cell liquid easily by with film
The contact free diffusing of the external solution of piece pincers, the normal activity of cell is compromised, and generally cell is utilizing patch-clamp
Will be progressively dead after two hours of progress Intracellular signals record.This also causes born of the same parents' patch clamp technique to turn into for a long time
One limiting factor of cell electric signal record.In addition, cell patchclamp technique is typically only capable to enter single nerve cell one by one
Horizontal electrical signal records, and very high to the technical requirements of experimenter, cannot enter horizontal electrical signal in multiple nerve cells with the time
Record, this strongly limits the communication understood between different nerve cells and Coordinating Activity.
Up to the present, most miniature neural interface is all built upon records nerve using the electrode of cell membrane exterior
The outer electric signal of epicyte.The outer electric signal of nerve cell is recorded due to weak output signal, the Asia of endocellular electricity signal cannot be recorded
Threshold value behavior, and easily interfered by cytoproximal extracellular electric signal is faced, endocellular electricity signal can not be carried out
Accurate record.In recent years, emerging nano-fabrication technique is rapidly developed novel micro nanometer device, between cell and electrode
Be electrically coupled be improved significantly so that the recording technique of extracellular electric signal is improved, but cannot still overcome cell dispatch from foreign news agency
Letter record is compared to obvious inferior position existing for endocellular electricity signal record.
Extracellular signal recording method, be non-invasive such as multiple electrode array and multi-transistor array, it is allowed to it is long-term and
Multiplexing measurement.However, extracellular recording sacrifices the one-to-one relationship between cell and electrode, and make signal intensity and matter
Amount significantly reduces.Therefore, extracellular technology can not be recorded with the action potential for exploring the degree of accuracy needed for ion channel property.Carefully
Intracellular recording method such as patch clamp technique enter cell interior to measure the voltage or electric current on cell membrane by using electrode, it is allowed to
With the amplitude and shape of high s/n ratio operation of recording current potential.However, intracellular process generally will record time limit due to invasive
System is in several hours, and its complexity makes it difficult to record multiple cells simultaneously.
Therefore, research and develop it is a kind of can at the same time it is interior multiple cells are entered horizontal electrical signal record device it is extremely urgent.
The content of the invention
The purpose of the present invention is overcome the deficiencies in the prior art, and specific one kind passes through nano-electrode An arrayed recording endocellular electricity
The device of signal, the device simply and easily can enter horizontal electrical signal record to multiple cells interior at the same time, in addition, it can
To record extracellular electric signal and endocellular electricity signal according to being actually needed, and except record nerve cell electric signal it
Outside, it can be also used for recording the electric signal of cardiac muscle cell or other kinds of cell.
In order to reach above-mentioned technical purpose, the present invention is realized by following technical scheme：
A kind of device by nano-electrode An arrayed recording endocellular electricity signal of the present invention, including silicon materials substrate
With the nano-electrode array being placed on silicon materials substrate, the position of nano-electrode array uses photoetching and reaction on silicon materials substrate
Ion(ic) etching is defined, and the position of non-nano electrod-array is covered by insulating properties silica on silicon materials substrate, institute
State nano-electrode array be some settings nano-electrode, the nano-electrode array with for stimulate/measure electronic component it
Between electrically connect.
As further getting close to for above-mentioned technology, the nano-electrode includes the silicon core of vertical, in the silicon core under
Portion is wrapped up by insulating properties shell, and the middle upper top of the silicon core is the conducting metal of biocompatibility.
As the further improvement of above-mentioned technology, the insulating properties shell of the middle and lower part of the silicon core is exhausted for biocompatibility
Edge oxide aluminium dioxide, insulating properties shell are the effects in order to prevent electric current by nano-electrode sidewall leakage.
As the further improvement of above-mentioned technology, the conducting metal of upper top is platinum electrode or gold electricity in the silicon core
Pole, silicon core and platinum electrode or gold electrode provide electric pathway for cell interior, for providing electro photoluminescence to cell or passing through electrochemistry
Reaction detection intracellular potential change.
In the present invention, the nano-electrode array is the nano-electrode of the setting of 4 × 4 array arrangements, the nanometer electricity
A diameter of 100-300nm of pole, its length are 1-3 microns, and the spacing of two adjacent nano-electrodes is 1-5 microns in array.
In the present invention, the nano-electrode array is distributed in block array, and block is shaped as square or circle or three
Angular, the area of the nano-electrode array is not more than 40x40 square microns, and the area of the nano-electrode array is less than typically
The size of neurocyte, so can be used in entering individual cells horizontal electrical signal record.
Compared with prior art, the beneficial effects of the invention are as follows：
(1) in the present invention, signal record in nerve cell can be used for by nano-electrode array and stimulate in culture dish
Cerebral Cortex neuron activity, so as to for studying multiple single Synaptic junctions.
(2) in the present invention, nano-electrode non-destructively can provide endocellular electricity to cell and stimulate, and record neuron
The endocellular electricity signal of cell, neuron generation action can be induced the pulse of neuron Injection Current by vertical nano-electrode
Current potential, these caused IAP changes can be recorded by membrane elastic property simultaneously to be verified；Anti- mistake
Come, with the operation of electric current patch-clamp pattern to Injection Current in nerve cell, by vertical nanowires electrod-array can monitor by
Each action potential caused by patch-clamp Injection Current.
(3) in the present invention, intracellular recording generally carries out signal and amplifies 100 times, and extracellular recording electric signal is amplified
For 1000 times, sample rate 5-100kHz, signal 1Hz-20KHz bandpass filtering, it records electric signal result and accurately may be used
Lean on.
Brief description of the drawings
The present invention is described in detail with specific embodiment below in conjunction with the accompanying drawings：
Fig. 1 is the structural representation of the device of the present invention by nano-electrode An arrayed recording endocellular electricity signal.
Fig. 2 is heretofore described nano pole structure schematic diagram.
Embodiment
As shown in figure 1, a kind of device by nano-electrode An arrayed recording endocellular electricity signal of the present invention, including
Silicon materials substrate 1 and the nano-electrode array 2 being placed on silicon materials substrate 1, correspond to nano-electrode battle array on silicon materials substrate 1
The position of row 2 is defined using photoetching and reactive ion etching, the placement of the non-nano electrod-array 2 on silicon materials substrate 1
Position is covered by insulating properties silica, the nano-electrode array 2 be some settings nano-electrode 21, the nanometer
Electrod-array 2 is electrically connected between electronic component 3 with for stimulating/measuring.
As shown in Fig. 2 the nano-electrode 21 includes the silicon core 211 of vertical, the middle and lower part of the silicon core 21 is by insulating
Property shell 212 wrap up, the middle upper top of the silicon core 21 is the conducting metal 213 of biocompatibility.Under in the silicon core 211
The insulating properties shell 212 in portion is biocompatibility insulation oxide aluminium dioxide, and insulating properties shell 212 is to prevent electric current from leading to
Cross the effect of nano-electrode sidewall leakage, the conducting metal 213 of upper top is platinum electrode or gold electrode in the silicon core 211, silicon
Core and platinum electrode or gold electrode provide electric pathway for cell interior, for providing electro photoluminescence to cell or being visited by electrochemical reaction
Survey intracellular potential change.
In the present invention, the nano-electrode array 2 for 4 × 4 array arrangements setting nano-electrode 21, the nanometer
A diameter of 100-300nm of electrode 21, its length are 1-3 microns, and the spacing of two adjacent nano-electrodes 21 is 1- in array
5 microns.
In the present invention, the nano-electrode array 2 is distributed in block array, and block is shaped as square or circle or three
Angular or other geometries, the area of the nano-electrode array 2 are not more than 40x40 square microns, the nano-electrode battle array
The area of row 2 is less than the size of general neurocyte, therefore it can be used in entering individual cells horizontal electrical signal record.
The process of detailed description below recording device records endocellular electricity signal of the present invention：
(1) culture hole for cultivating cell He holding cell culture fluid is made using dimethyl silicone polymer.
(2) by dimethyl silicone polymer culture hole with nano-electrode array table is mutually assembled, nano-electrode array 2 and thorn
The connection for swashing/measuring between electronic component is realized using the printed circuit board (PCB) of conductive elastic connector and customization.
(3) cell 10 is incubated on nano-electrode array 2, cell membrane is penetrated by nano-electrode array 2 so that receives
Rice electrode can directly contact the cytokine milieu in cell 10, after cell starts beating, be amplified using 60 channel voltages
The electric signal (each 16 nano-electrodes of array) of cell of the device system record culture on nano-pillar electrod-array, in identical
By the use of Ag/AgCl electrodes in the medium as reference electrode in culture medium, intracellular recording generally carries out signal and amplifies 100 times,
Extracellular recording electric signal is amplified as 1000 times, sample rate 5-100kHz, signal 1Hz-20KHz bandpass filtering.
The present invention is specifically described below by way of several embodiments：
Embodiment one
The mouse neuronal cell line cultivated on nano-electrode array 2 shows normal growth, and microscope imaging shows, nanometer
The nerve cell grown on electrod-array 2 shows the morphological cellular similar to plane domain in rhythmicity hopping process, will
After the overcritical freeze-drying of cell progress is fixed, it is observed using SEM (SEM), shows nerve growth
On nano-electrode.
Embodiment two
Using with the action potential recorded on nano-electrode array 2 in nerve cell.The signal intensity detected and electricity
The detection zone of pole is directly related, is operated with electric current patch-clamp pattern to Injection Current in nerve cell, passes through nano-electrode battle array
Row 2 can monitor the endocellular electricity recorded as each action potential caused by patch-clamp Injection Current, nano-electrode array 2
The signal to noise ratio of signal is typically larger than 100.
Embodiment three
Continuous repetition in several days records multiple endocellular electricity signals on nano-electrode array 2, can observe and record simultaneously
Action potential change in different nerve cells, these nerve cells contact with each other, and the communication that contacts with each other are formed, when to wherein one
After individual nerve cell carries out electro photoluminescence, it can be observed in other nerve cells and produce corresponding intracellular potential change.
The invention is not limited in above-mentioned embodiment, every various changes or modifications to the present invention do not depart from the present invention
Spirit and scope, if these changes and modification belong within the scope of the claim and equivalent technologies of the present invention, then this hair
It is bright to also imply that comprising these changes and modification.
Claims (6)
- A kind of 1. device by nano-electrode An arrayed recording endocellular electricity signal, it is characterised in that：Including silicon materials substrate and The nano-electrode array being placed on silicon materials substrate, on silicon materials substrate the position of nano-electrode array using photoetching and react from Son etching is defined, and the position of non-nano electrod-array is covered by insulating properties silica on silicon materials substrate, described Nano-electrode array is the nano-electrode of some settings, the nano-electrode array and for stimulating/measuring between electronic component Electrical connection.
- 2. the device according to claim 1 by nano-electrode An arrayed recording endocellular electricity signal, it is characterised in that：Institute Stating nano-electrode includes the silicon core of vertical, and the middle and lower part of the silicon core is wrapped up by insulating properties shell, upper in the silicon core to push up Portion is the conducting metal of biocompatibility.
- 3. the device according to claim 1 by nano-electrode An arrayed recording endocellular electricity signal, it is characterised in that：The insulating properties shell of the middle and lower part of the silicon core is biocompatibility insulation oxide aluminium dioxide.
- 4. the device according to claim 1 by nano-electrode An arrayed recording endocellular electricity signal, it is characterised in that：The conducting metal of upper top is platinum electrode or gold electrode in the silicon core.
- 5. the device by nano-electrode An arrayed recording endocellular electricity signal according to any one of Claims 1-4, it is special Sign is：The nano-electrode array for 4 × 4 array arrangements setting nano-electrode, the nano-electrode it is a diameter of 100-300nm, its length are 1-3 microns, and the spacing of two adjacent nano-electrodes is 1-5 microns in array.
- 6. the device according to claim 5 by nano-electrode An arrayed recording endocellular electricity signal, it is characterised in that：Institute State nano-electrode array to be distributed in block array, block is shaped as square or circle or triangle, the nano-electrode array Area be not more than 40x40 square microns.
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| CN111289819A (en) * | 2020-04-09 | 2020-06-16 | 中山大学 | Integrated recording regulation and control system for measuring intracellular electric signals by myocardial cell electroporation |
| CN114137191A (en) * | 2021-12-29 | 2022-03-04 | 上海交通大学 | Multifunctional electrode array system for cell biochemical signal detection and regulation |
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| CN114137191A (en) * | 2021-12-29 | 2022-03-04 | 上海交通大学 | Multifunctional electrode array system for cell biochemical signal detection and regulation |
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