CN101639474B - Device for detecting three-dimensional growth of nerve cells and electric signals thereof - Google Patents
Device for detecting three-dimensional growth of nerve cells and electric signals thereof Download PDFInfo
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Abstract
The invention discloses a device for detecting three-dimensional growth of nerve cells and electric signals thereof, which is characterized in that more than two layers of cell growing devices are fixedly connected between two layers of microelectrode array sensors, each cell growing device is provided with microchambers which are arranged in a matrix mode, the corresponding microchambers of the cell growing devices at the different layers are opposite up and down, and every two adjacent microchambers on the same line and the same row of each cell growing device are communicated by a microchannel; a large microelectrode is correspondingly arranged in each of the microchambers of the cell growing devices at the bottom layer and the top layer, and a small microelectrode is correspondingly arranged in each microchannel; a communicating vessel is arranged between two layers of cell growing devices and is provided with a micro through hole matrix corresponding to the upper microchamber and the lower microchamber; a channel is formed among the microchambers which are mutually corresponding up and down and the micro through holes of the communicating vessel; the size of each microchamber only allows to contain one nerve cell; and the width of each microchannel only allows to contain one nerve synapse. The device ensures that the cultured nerve cells can be sequentially connected in the three-dimensional space, and can detect the electric signals of the nerve cells.
Description
Technical field
The present invention relates to detect the device of electrical signal of nerve cell, particularly use the device of microelectrode array sensor electrical signal of nerve cell.
Background technology
The analysis of electroneurographic signal is one of the main contents of the research of present nuroinformatics.Device now relatively more commonly used be the microelectrode array sensor (Micro-Electrode Array, MEA).MEA is made up of the microelectrode that is embedded on the substrate.Mainly for example platinum, gold, titanium nitride and indium tin oxide etc. constitute microelectrode by metal material.Microelectrode is lined up array on substrate (normally glass).Microelectrode links to each other with the lead of processing with gold or transparent indium tin oxide, and lead is sent to outside amplifier with the microelectrode signal, or the stimulation that the outside applies is sent to microelectrode.In MEA equipment, tried cell or tissue directly on the electrode coating material, to cultivate, the local electricity that can allow to measure the extracellular direct neighbor changes the electricity physiological signal of while and a plurality of cells of line item.Research based on the Neurobiology of external MEA technology has two outstanding advantages, and the one, can write down simultaneously and stimulate different loci, the 2nd, non-intrusion type can damaging cells.On same culture sample, grow the time-histories record so MEA is applicable to, thereby allow to keep watch on the differentiation of culture the long period of the response of stimulation.But there are two intrinsic bottlenecks in existing MEA technology: the one, can't make the cell directional growth; Connect in the cynapse of the neurocyte of in vitro culture and to be at random; With a great difference that is connected with of cynapse in the actual biosome, and in vitro culture each time, the connection of neurocyte also is different.And the connected mode of the electric signal of each neuron granting and this neuron and peripheral nerve unit has much relations.Existing this external connected mode at random not only lets the experiment of nervous physiology can't have accurate repeatability, and can't obtain topological structure and nerve signal corresponding relation that neurocyte connects; The 2nd, under MEA planar micro electrode structure, neurocyte can only be in the growth of MEA substrate surface, so the topological structure of the nerve synapse of interconnected growth is limited on the MEA surface on the script space.The network structure that under this condition, obtains does not conform to through the network that being connected to form of nerve synapse has the space multistory structure with neurocyte in the actual biosome.Therefore; To a certain extent; Current with the MEA technology be the electrophysiological scientific research of neurocyte carried out of basis with biosome in the neural network that is connected in order of real space be discrepant, present plane culture technique has restricted the electrophysiological development of neurocyte.
Summary of the invention
Technical matters to be solved by this invention provides a kind of may command neurocyte connects and obtain its electric signal in order in solid space device.
For the neurocyte that overcomes in vitro culture in the prior art connects this shortcoming in the plane at random; Inventive concept of the present invention is: two-layer above cell growth assembly is set; Each microchamber of cell growth assembly is arranged by matrix-style; And the microchannel of connection is set between each microchamber with the cell growth assembly of one deck, then be communicated with between each confluent monolayer cells grower, and form passage between the micro through hole of microchamber, the linker of microchamber, the top layer cell growth assembly of corresponding bottom cell growth assembly each other through linker; Thereby make and in space multistory, to connect into network according to specific mode in cultured cells on the MEA; The electric signal of the microelectrode recording cell through MEA can detect the electric signal of the cell under the specific three-dimensional connected mode like this, obtains the relation of cell electric signal and its connected mode.The number of plies of cell growth assembly and the arrangement mode of microchamber in the cell growth assembly and microchannel can be according to the connected mode settings of neurocyte in the biosome and nerve synapse; Cultivate the network of the neurocyte formation on this device like this and can simulate the neural network in the actual biosome; Therefore detected electroneurographic signal approaches real electroneurographic signal in the biosome, thereby detected electroneurographic signal can comprise a large amount of biological informations.The connected mode setting of the neural network that the number of plies of cell growth assembly and the arrangement mode of microchamber in the cell growth assembly and microchannel also can be wanted to create according to people; The neurocyte of cultivating like this on this device just can connect according to the connected mode that people need; Thereby can obtain three-dimensional artificial neural network, the microelectrode of MEA can come out the electrical signal detection of this artificial neural network.
In order to solve the problems of the technologies described above, technical scheme of the present invention is following:
The device of three-dimensional growth of nerve cells of the present invention and electrical signal detection thereof mainly comprises bottom microelectrode array sensor, top layer microelectrode array sensor, bottom cell growth assembly, top layer cell growth assembly and linker; Said bottom cell growth assembly and top layer cell growth assembly are located between bottom microelectrode array sensor and the top layer microelectrode array sensor; Said bottom microelectrode array sensor and top layer microelectrode array sensor comprise substrate, are located at said suprabasil big microelectrode and little microelectrode; The substrate of said bottom cell growth assembly and bottom microelectrode array sensor is fixedly connected; The substrate of said top layer cell growth assembly and top layer microelectrode array sensor is fixedly connected; Said bottom cell growth assembly and top layer cell growth assembly include the microchamber of arranging by matrix-style; The corresponding microchamber of the microchamber of said bottom cell growth assembly and top layer cell growth assembly is relative up and down, all has a microchannel to be communicated with between the adjacent microchamber of the same delegation of said bottom cell growth assembly and top layer cell growth assembly and between the adjacent microchamber of same row; Be equipped with a big microelectrode of said bottom microelectrode array sensor in the microchamber of said bottom cell growth assembly accordingly, the little microelectrode of said bottom microelectrode array sensor is positioned at the microchannel of said bottom cell growth assembly accordingly; Be equipped with a big microelectrode of said top layer microelectrode array sensor in the microchamber of said top layer cell growth assembly accordingly, the little microelectrode of said top layer microelectrode array sensor is positioned at the microchannel of top layer cell growth assembly accordingly; Said linker is located between bottom cell growth assembly and the top layer cell growth assembly; And said linker and bottom cell growth assembly and top layer cell growth assembly are fixedly connected; Be provided with in the said linker and the microchamber of said bottom cell growth assembly and corresponding up and down the micro through hole of microchamber of top layer cell growth assembly by the matrix-style arrangement; Form passage between the micro through hole of the microchamber of the bottom cell growth assembly of said mutual correspondence, the microchamber of top layer cell growth assembly, linker; The size of said microchamber is for only holding a neurocyte, and the width of said microchannel is for only holding a nerve synapse.
Further; Linker according to the invention has two; Said two linkers are fixedly connected with said top layer cell growth assembly and bottom cell growth assembly respectively; The intermediate layer cell grower also has been fixedly connected between said two linkers; Said intermediate layer cell grower is provided with and the microchamber of said bottom cell growth assembly and the corresponding microchamber by the matrix-style arrangement of microchamber of top layer cell growth assembly; All there is the microchannel to be communicated with between the adjacent microchamber of the same delegation of said intermediate layer cell grower and between the adjacent microchamber of same row, forms passage between the microchamber of the microchamber of the bottom cell growth assembly of said mutual correspondence, the micro through hole of linker, intermediate layer cell grower, the microchamber of top layer cell growth assembly.
Compared with prior art; Advantage of the present invention is: (1) is because the present invention is provided with the cell growth assembly of the different numbers of plies; Cell growth assembly is provided with microchamber and microchannel again; Make to cultivate and to carry out oriented growth with the neurocyte in the confluent monolayer cells grower according to the arrangement mode of microchamber and microchannel, the cell in the cell growth assembly of different layers connects through micro through hole, thereby the neurocyte of cultivation can connection in order in solid space in this device; (2) arrangement mode of the number of plies of cell growth assembly and microchamber in the cell growth assembly and microchannel can be according to the connected mode setting of neurocyte in the biosome and nerve synapse; Cultivate the network of the neurocyte formation on this device like this and can simulate the neural network in the actual biosome; Microelectrode through in microchamber and the microchannel detects electroneurographic signal; Therefore detected electroneurographic signal approaches real electroneurographic signal in the biosome, thereby detected electroneurographic signal can comprise a large amount of biological informations.(3) the connected mode setting of the arrangement mode of the number of plies of cell growth assembly and microchamber in the cell growth assembly and microchannel and the neural network that also can want to create according to people; The neurocyte of cultivating like this on this device just can connect according to the connected mode that people need; Thereby can obtain artificial neural network, microelectrode can come out the electrical signal detection of this artificial neural network.
Description of drawings
Fig. 1 is the structural representation of first kind of embodiment of apparatus of the present invention;
Fig. 2 is the A-A cut-open view of Fig. 1;
Fig. 3 is the B-B cut-open view of Fig. 1;
Fig. 4 is the structural representation of second kind of embodiment of apparatus of the present invention;
Fig. 5 is the A-A cut-open view of Fig. 4;
Fig. 6 is the B-B cut-open view of Fig. 4;
Fig. 7 is the structural representation of cell growth assembly of the present invention;
Fig. 8 is the D-D cut-open view of Fig. 7;
Fig. 9 is the structural representation of connector of the present invention;
Figure 10 is the C-C cut-open view of Fig. 9;
Figure 11 is the structural representation of microelectrode array sensor of the present invention;
Figure 12 is the F-F cut-open view of Figure 11.
Embodiment
Fig. 1, Fig. 2 and Fig. 3 show the structure of first kind of embodiment of apparatus of the present invention.This device comprises bottom cell growth assembly 2, top layer cell growth assembly 1, bottom microelectrode array sensor 42, top layer microelectrode array sensor 41 and linker 3.Bottom cell growth assembly 2 includes the microchamber of arranging by matrix-style with top layer cell growth assembly 1.Below illustrate, as bottom cell growth assembly 2 and top layer cell growth assembly 1 as Fig. 1 and shown in Figure 7 when comprising nine microchambers, nine microchambers are according to the matrix-style arrangement of three rows, three row.The corresponding microchamber of the microchamber of bottom cell growth assembly 2 and top layer cell growth assembly 1 is relative up and down, so that the cynapse of cultivating in the neurocyte in the microchamber of neurocyte and bottom cell growth assembly 2 in the microchamber of top layer cell growth assembly 1 can be connected with each other.As a kind of preferred implementation wherein; In the top layer cell growth assembly 1 microchamber be positioned at the bottom cell growth assembly 2 relative with it microchamber directly over; Make the central lines of microchamber of microchamber and the bottom cell growth assembly 2 of relative up and down top layer cell growth assembly 1, guarantee that the neurocyte in the corresponding microchamber of different layers can interconnect.All there is a microchannel to be communicated with between the adjacent microchamber of the same delegation of bottom cell growth assembly 2 and top layer cell growth assembly 1 and between the adjacent microchamber of same row: specifically; For example like Fig. 1 and shown in Figure 7; In top layer cell growth assembly 1; Be positioned at the microchamber with delegation: first microchamber 12 and second microchamber 13 are communicated with through first microchannel 5, and the 3rd microchamber 14 and the 4th microchamber 15 are communicated with through the 3rd microchannel 7; Second microchamber 13 that is positioned at same row is communicated with through second microchannel 6 with the 3rd microchamber 14, and the 4th microchamber 15 is communicated with through the 4th microchannel 8 with first microchamber 12; Connection between other microchamber by that analogy.Mode of communicating between microchamber in the bottom cell growth assembly 2 and the microchannel is identical with the mode of communicating between the microchannel with the microchamber of top layer cell growth assembly 1.Like Fig. 2, Fig. 3, Figure 11 and shown in Figure 12, bottom microelectrode array sensor 42 comprises substrate 4, is located at big microelectrode 9 and little microelectrode 10 in the substrate 4 with top layer microelectrode array sensor 41.Like Fig. 1, Fig. 2 and shown in Figure 3; Bottom cell growth assembly 2 is fixedly connected with the substrate 4 of bottom microelectrode array sensor 42; The big microelectrode 9 of bottom microelectrode array sensor 42 is positioned at the microchamber of bottom cell growth assembly 2 accordingly; Each microchamber that is bottom cell growth assembly 2 is built-in with a big microelectrode 9 on the bottom microelectrode array sensor 42; The little microelectrode 10 of bottom microelectrode array sensor 42 is positioned at the microchannel of bottom cell growth assembly 2 accordingly, promptly can be equipped with two little microelectrodes 10 on the bottom microelectrode array sensor along length a direction in each microchannel of bottom cell growth assembly 2.Top layer cell growth assembly 1 is fixedly connected with the substrate 4 of top layer microelectrode array sensor 41; Each microchamber of top layer cell growth assembly 1 is built-in with a big microelectrode 9 of top layer microelectrode array sensor 41; The little microelectrode 10 of top layer microelectrode array sensor 41 is positioned at the microchannel of top layer cell growth assembly 1 accordingly, and the corresponding relation of microchamber in the corresponding relation of microchamber in the top layer cell growth assembly 1 and big microelectrode 9 and the corresponding relation of microchannel and little microelectrode 10 and the bottom cell growth assembly 2 and microelectrode 9 greatly and the corresponding relation of microchannel and little microelectrode 10 are identical.The linker 3 that has been fixedly connected between the microchamber of bottom cell growth assembly and the top layer cell growth assembly is provided with in the linker 3 and the microchamber of bottom cell growth assembly 2 and the corresponding up and down micro through hole 11 by the matrix-style arrangement of microchamber of top layer cell growth assembly 1.Like Fig. 2, Fig. 3, Fig. 9 and shown in Figure 10; Nine micro through holes 11 are arranged in the linker 3; Nine micro through holes are arranged according to the matrix-style of three rows, three row; The microchamber of the position of micro through hole 11 and bottom cell growth assembly 2 and the microchamber of top layer cell growth assembly 1 are corresponding; Be that micro through hole 11 is accordingly between the microchamber of the microchamber of relative up and down bottom cell growth assembly 2 and top layer cell growth assembly 1; Each other form passage between the micro through hole 11 of the microchamber of the microchamber of corresponding bottom cell growth assembly 2, top layer cell growth assembly 1, linker 3 up and down, make cynapse in the neurocyte in the microchamber of cultivating neurocyte and bottom cell growth assembly 2 in the microchamber of top layer cell growth assembly 1 can put in the micro through hole 11 and couple together, realized neurocyte ordering growth in solid space.If like Fig. 1, Fig. 3, Fig. 4 and shown in Figure 6; Sidewall of micro through hole 11 and sidewall with its relative microchamber up and down are positioned on the same plane, can guarantee better then that up and down nerve synapse in two microchambers is pasting sidewall and putting in the micro through hole and connect.
The big I of the microchamber of the microchamber of bottom cell growth assembly 2 and top layer cell growth assembly 1 equals or is slightly larger than the diameter of a neurocyte, makes the size of each microchamber only can hold a neurocyte; And some short nerve synapses possibly can not stretched in the microchannel.
As shown in Figure 1, the width b of each microchannel is for only holding a nerve synapse.The purpose of doing like this is: the electric signal on nerve synapse is only detected by the little microelectrode in the microchannel; Little microelectrode in microchannel only can detect the electric signal of a nerve synapse, makes that detected electric signal of little microelectrode and the nerve synapse in the microchannel forms one-to-one relationship.
As shown in figures 1 and 3; The length c of micro through hole 11 and width d are the diameter greater than a nerve synapse; Guaranteeing that the cynapse of the neurocyte in the corresponding microchamber can put in the micro through hole 11 in cynapse and the bottom cell growth assembly 2 of neurocyte in the microchamber in the top layer cell growth assembly 1 interconnects; Through changing the length c and the width d of micro through hole 11; Can control the quantity that puts in the nerve synapse in the micro through hole, thus the neurocyte in the control top layer cell growth assembly 1 and the strength of joint of the neurocyte in the bottom cell growth assembly 2.
The diameter of big microelectrode 9 preferably is about the diameter of the neurocyte that institute will detect, and the purpose of doing like this is: both made big microelectrode ability structure be arranged in microchamber fully, can detect the electric signal of neurocyte again.
The diameter of little microelectrode 10 preferably with the nerve synapse that will detect equal diameters or be slightly less than the nerve synapse diameter, this is in order both to make little microelectrode be arranged in the microchannel fully by structure, to detect the electric signal of nerve synapse again.
More than provided first kind of embodiment of the device of the three-dimensional growth of nerve cells that comprises two-layer cell growth assembly and electrical signal detection thereof.The present invention can be on the basis of first kind of embodiment; Obtain the two-layer above three-dimensional growth of nerve cells and the device of electrical signal detection thereof through the number that increases cell growth assembly and linker; Introduce second kind of embodiment of apparatus of the present invention below, this second kind of embodiment has three confluent monolayer cells growers and two linkers.
Like Fig. 1 and shown in Figure 4, the vertical view of second kind of embodiment of the present invention and first kind of embodiment is the same.Like Fig. 5 and shown in Figure 6; The linker of second kind of embodiment of the present invention has two; It is identical with the structure of second linker 32 promptly to be equipped with micro through hole 11, the first linkers 31 in first linker 31 and second linker, 32, the first linkers 31 and second linker 32.Wherein, first linker 31 is fixedly connected with top layer cell growth assembly 1, and second linker 32 is fixedly connected with bottom cell growth assembly 2.Intermediate layer cell grower 15 has been fixedly connected between first linker 31 and second linker 32.Like Fig. 7 and shown in Figure 8; Intermediate layer cell grower 15 is identical with the structure of top layer cell growth assembly 1 and bottom cell growth assembly 2; Be that intermediate layer cell grower 15 also is provided with and the microchamber of bottom cell growth assembly 2 and the identical microchamber by the matrix-style arrangement of microchamber of top layer cell growth assembly 1 accordingly, and all have a microchannel to be communicated with between each adjacent microchamber of the same delegation of intermediate layer cell grower 15 and between each adjacent microchamber of same row.Form a passage up and down each other between the microchamber of the micro through hole 11 of the microchamber of the micro through hole 11 of the microchamber of corresponding bottom cell growth assembly 2, second linker 32, intermediate layer cell grower 15, first linker 31, top layer cell growth assembly 1; The cynapse of make cultivating neurocyte and the neurocyte in the intermediate layer cell grower 15 in top layer cell growth assembly 1 can put in the micro through hole 11 of first linker 31 and couple together; Cynapse in the neurocyte in neurocyte in the intermediate layer cell grower 15 and the bottom cell growth assembly 2 can put in the micro through hole 11 of second linker 32 and couple together, and has realized neurocyte ordering growth in solid space.
In second kind of embodiment of the present invention, the requirement of microchamber, microchannel, big microelectrode 9, little microelectrode 10 and the size of micro through hole 11 is identical with first kind of embodiment.
The method of application of apparatus of the present invention:
For the three-dimensional growth of nerve cells of first kind of embodiment of the present invention and the device of electrical signal detection thereof, being fixedly connected between top layer cell growth assembly 1, linker 3 and the bottom cell growth assembly 2 is detachable.During use; The bottom cell growth assembly 2 that at first will be fixedly connected with bottom microelectrode array sensor 42, the top layer cell growth assembly 1 and the linker 3 that have been fixedly connected with top layer microelectrode array sensor are separated; Cultured cell on the big microelectrode 9 in the different microchambers of top layer cell growth assembly 1 and bottom cell growth assembly 2 respectively, institute's cultured cells is attached on the big microelectrode 9.Two days later; Neurocyte on top layer cell growth assembly 1 and the bottom cell growth assembly 2 grows cynapse along the microchannel; Cynapse is attached on the little microelectrode 10; At this moment; In a double dish that nutrient solution is housed; Top layer cell growth chamber 1, linker 3 and bottom cell growth assembly 2 are fixedly connected, notice that nutrient solution will not have top layer cell growth assembly 1, then in double dish with face of the whole upset of the device of three-dimensional growth of nerve cells of the present invention and electrical signal detection thereof; The top layer cell growth assembly 1 and the bottom cell growth assembly 2 of original like this two positions up and down that are in vertical direction is positioned at former and later two positions of horizontal direction now, so the cynapse of the neurocyte of top layer cell growth assembly 1 and the bottom cell growth assembly 2 of former and later two positions can easily put in the micro through hole and couples together on the horizontal direction.Big microelectrode 9 on top layer microelectrode array sensor 41 and the bottom microelectrode array sensor 42 can detect the electric signal that adheres to neurocyte above that, and little microelectrode 10 can detect the electric signal that adheres to nerve synapse above that.
For the three-dimensional growth of nerve cells of second kind of embodiment of the present invention and the device of electrical signal detection thereof; During use; At first the top layer cell growth assembly 1 and first linker 31 are separated; First linker 31 is separated with intermediate layer cell growth 15 devices, and second linker 32 is separated with bottom cell growth assembly 2.But second linker 32 is not separated with intermediate layer cell grower 15, because top layer cell growth assembly 1 has been fixed in the MEA substrate with bottom cell growth assembly 2, can cultured neurons; And intermediate layer cell grower 15 is not owing to be fixed in the MEA substrate, and its lower end is uncovered, can't cultured cell, therefore need intermediate layer cell grower 15 be fixed on second linker 32.Top layer cell growth assembly 1, bottom cell growth assembly 2, first linker 31 and second linker 32 are placed in the double dish that nutrient solution is housed; On the wall with second linker 32 of cellular incubation in intermediate layer cell grower 15; Cell can wall attached to second linker 32 on; On the big microelectrode 9 of cellular incubation in the 2 different microchambers put are adorned in the growth of top layer cell growth assembly 1 and bottom cell, cell can be attached on the big microelectrode 9, two days later; Neurocyte grows cynapse along the microchannel; Cynapse is attached to little microelectrode 10, and wherein the cynapse in the intermediate layer cell grower 15 is only grown in the microchannel of intermediate layer cell grower 15, can't be attached on the little microelectrode.At this moment, top layer cell growth chamber device 1, first linker 31, intermediate layer cell grower 15, second linker 32 and bottom cell growth assembly 2 are fixedly connected, notice that nutrient solution will not have the top layer cell growth assembly.Then in double dish with face of the whole upset of the device of three-dimensional growth of nerve cells of the present invention and electrical signal detection thereof; Original like this top layer cell growth assembly 1, intermediate layer cell grower 15 and bottom cell growth assembly 2 that is in vertical direction is positioned at horizontal direction now; Therefore the cynapse of the neurocyte in the top layer cell growth assembly on the horizontal direction 1 and the intermediate layer cell grower 15 can easily put in the micro through hole of first linker 31 and couple together, and the cynapse of the neurocyte of intermediate layer cell grower 15 and bottom cell growth assembly 2 can easily put in the micro through hole of second linker 32 and couple together.Big microelectrode 9 on top layer microelectrode array sensor 41 and the bottom microelectrode array sensor 42 can detect the electric signal that adheres to neurocyte above that, and little microelectrode 10 can detect the electric signal that adheres to nerve synapse above that.
More than be two kinds of embodiments of the present invention, in practical application, can according to the quantity of the cell that will detect the quantity of microchamber is set; Neurocyte arrangement and ways of connecting according to different in the biosome are provided with the number of plies of cell growth chamber and the arrangement mode of microchamber and microchannel; Thereby the connected mode of the neurocyte of on apparatus of the present invention, cultivating can be simulated the connected mode of the neurocyte in the biosome; Therefore the electric signal of the electric signal of the neurocyte of cultivation on this device and the neurocyte in the biosome is similar, and the electric signal that measures has carried more biological informations; Also the number of plies of cell growth chamber and the arrangement mode of microchamber and microchannel can be set according to the connected mode of the artificial neural network of wanting to create; Will connect into the artificial neural network of wanting thereby this device is gone up cultured cells, obtain the electric signal of artificial neural network through the microelectrode detection of MEA.
Claims (2)
1. the device of three-dimensional growth of nerve cells and electrical signal detection thereof; It is characterized in that: it comprises bottom microelectrode array sensor, top layer microelectrode array sensor, bottom cell growth assembly, top layer cell growth assembly and linker; Said bottom cell growth assembly and top layer cell growth assembly are located between bottom microelectrode array sensor and the top layer microelectrode array sensor; Said bottom microelectrode array sensor and top layer microelectrode array sensor comprise substrate, are located at said suprabasil big microelectrode and little microelectrode; The substrate of said bottom cell growth assembly and bottom microelectrode array sensor is fixedly connected; The substrate of said top layer cell growth assembly and top layer microelectrode array sensor is fixedly connected; Said bottom cell growth assembly and top layer cell growth assembly include the microchamber of arranging by matrix-style; The corresponding microchamber of the microchamber of said bottom cell growth assembly and top layer cell growth assembly is relative up and down, all has a microchannel to be communicated with between the adjacent microchamber of said bottom cell growth assembly and top layer cell growth assembly same delegation separately and between the adjacent microchamber of same row, does not intersect between each microchannel; Be equipped with a big microelectrode of said bottom microelectrode array sensor in the microchamber of said bottom cell growth assembly accordingly, the little microelectrode of said bottom microelectrode array sensor is positioned at the microchannel of said bottom cell growth assembly accordingly; Be equipped with a big microelectrode of said top layer microelectrode array sensor in the microchamber of said top layer cell growth assembly accordingly, the little microelectrode of said top layer microelectrode array sensor is positioned at the microchannel of top layer cell growth assembly accordingly; Said linker is located between bottom cell growth assembly and the top layer cell growth assembly; And said linker and bottom cell growth assembly and top layer cell growth assembly are fixedly connected; Be provided with in the said linker and the microchamber of said bottom cell growth assembly and corresponding up and down the micro through hole of microchamber of top layer cell growth assembly by the matrix-style arrangement; Form passage between the micro through hole of the microchamber of the bottom cell growth assembly of said mutual correspondence, the microchamber of top layer cell growth assembly, linker; The size of said microchamber is for only holding a neurocyte, and the width of said microchannel is for only holding a nerve synapse.
2. the device of three-dimensional growth of nerve cells according to claim 1 and electrical signal detection thereof; It is characterized in that: said linker has two; Said two linkers are fixedly connected with said top layer cell growth assembly and bottom cell growth assembly respectively; The intermediate layer cell grower also has been fixedly connected between said two linkers; Said intermediate layer cell grower is provided with and the microchamber of said bottom cell growth assembly and the corresponding microchamber by the matrix-style arrangement of microchamber of top layer cell growth assembly; All there is the microchannel to be communicated with between the adjacent microchamber of the same delegation of said intermediate layer cell grower and between the adjacent microchamber of same row, forms passage between the microchamber of the microchamber of the bottom cell growth assembly of said mutual correspondence, the micro through hole of linker, intermediate layer cell grower, the microchamber of top layer cell growth assembly.
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