CN107828652B - The cell activity adjusting method and device of Cell-oriented Activity determination experiment - Google Patents
The cell activity adjusting method and device of Cell-oriented Activity determination experiment Download PDFInfo
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Abstract
The present invention discloses the cell activity adjusting method and device of a kind of Cell-oriented Activity determination experiment, it is ultrasonic transducer right above electric cell-substrate impedance sensor, ultrasonic transducer is fixedly connected with the transducer height regulating device for adjusting ultrasonic transducer relative to distance between electric cell-substrate impedance sensor, electric cell-substrate impedance sensor detects controller through control line connection cell impedance, and ultrasonic transducer connects ultrasonic transducer controller through control line；PC host computer connects MCU controller by data line, and MCU controller is separately connected ultrasonic transducer controller, cell impedance detection controller and transducer height regulating device；The model of impedance value Yu irradiation distance and irradiation time is fitted with BP neural network, optimum combination is found offline with genetic algorithm, finally more accurately carry out cell activity on-line control, impedance error caused by reducing the cell death inducer that add various dose various concentration in cell culture fluid.
Description
Technical field
The present invention relates to cytoactive detection experimental techniques, specifically carry out cell activity in cytoactive detection experiment
The method and apparatus of adjusting.
Background technique
Currently, need to establish different cell activity grades according to different requirements, in cytoactive detection experiment,
It needs to automatically adjust to cell activity.A kind of use is disclosed in the document that Chinese Patent Application No. is 95116995.5
The method that cell activity is adjusted in non-toxic, a effective amount of form of hyaluronic acid (pharmaceutically acceptable salt), this germplasm
Acid is a kind of Sodium Hyaluronate that form is the bottled 20mg/mL of 15mL, this Sodium Hyaluronate is 2% solution, also contains three in bottle
It steams, aqua sterilisa.When carrying out cell activity adjusting in this way, need to be added dropwise the hyalomitome of various concentration into cell culture fluid
Sour sodium causes to will cause error to cell impedance value measured by electric cell-substrate impedance sensor in cytoactive detection experiment.Also
Have it is a kind of adjust cell activity method, be by add various dose cell death inducer induce cell apoptosis, thus
Generate different grades of cell activity index.Both methods requires the medicament time of about half an hour, tests
Step very complicated, and there are the factors of human interference.If a kind of method that cell activity is adjusted can be found, do not need
Any medicament is added, the experimental procedure of very complicated is not needed, it, can be by automatic as long as cell activity grade needed for input
The device of change realizes that cell activity is accurately adjusted, then will generate far-reaching influence to medicine, biology, cytology.
Summary of the invention
The purpose of the invention is to save the existing tedious steps with inducer of apoptosis preparation cytoactive detection sample
With the tediously long time, manual operation and inductive dose difference bring error are reduced, a kind of Cell-oriented Activity determination is proposed
The cell activity adjusting method and device of experiment are shone using low-intensity low frequency ultrasound amplitude, realize oneself of cell activity grade
Dynamicization is adjusted.
The cell activity regulating device of Cell-oriented Activity determination proposed by the present invention experiment the technical solution adopted is that: packet
Electric cell-substrate impedance sensor is included, is ultrasonic transducer right above electric cell-substrate impedance sensor, ultrasonic transducer is fixedly connected with tune
Save transducer height regulating device of the ultrasonic transducer relative to distance between electric cell-substrate impedance sensor；Electric cell-substrate impedance sensor
Controller is detected through control line connection cell impedance, ultrasonic transducer connects ultrasonic transducer controller through control line；PC
Host computer connects MCU controller by data line, and MCU controller is separately connected ultrasonic transducer controller, cell impedance inspection
Survey controller and transducer height regulating device.
The transducer height regulating device is made of stepper motor, controllor for step-by-step motor, sliding block and screw rod, stepping
Motor connects controllor for step-by-step motor through step motor control line, and controllor for step-by-step motor connects MCU controller；Stepper motor hangs down
Straight arrangement, output shaft are fixedly and coaxially connected the lower end of screw rod, and cooperation is cased with sliding block on screw rod, and sliding block is fixedly connected with ultrasonic waves
It can device.
The skill that the adjusting method of the cell activity regulating device of Cell-oriented Activity determination experiment proposed by the present invention uses
Art scheme be the following steps are included:
A, PC host computer controls transducer height adjustment device work through MCU controller, makes ultrasonic transducer to moving up
The home position of highest point is moved, ultrasound wave irradiation distance is h at this time1；Cell culture fluid, PC are added dropwise in electric cell-substrate impedance sensor
The ultrasound wave irradiation time of PC control ultrasonic transducer is t1, electric cell-substrate impedance sensor detects cell impedance value Z11；So
After replace cell culture fluid, the control ultrasound wave irradiation time is t2, detect cell impedance value Z12；It loops back and forth like this, until PC
The host computer acquisition ultrasound wave irradiation time is sequentially t1,t2,…,tnN sample data (h1, t1, Z11),(h1, t2,
Z12),…,(h1, tn, Z1n), first sampling period terminates；
B, PC PC control ultrasonic transducer moves down, and obtaining ultrasound wave irradiation distance is h2, in ultrasound wave irradiation distance h2No
Under conditions of change, the PC host computer acquisition ultrasound wave irradiation time is sequentially t1,t2,…,tnN sample data (h2, t1,
Z21),(h2, t2, Z22),…,(h2, tn, Z2n), second sampling period terminates；
C, and so on, obtain the sample data (h in m-th of sampling periodm, t1, Zm1),(hm, t2, Zm2),…,
(hm, tn, Zmn), total mn group data；
D, PC host computer is with ultrasound wave irradiation distance h1,h2,…,hm, irradiation time t1,t2,…,tnAs BP neural network
Input, with cell impedance value Z11,,…,ZmnAs output, BP neural network model Z=f (h, t) is obtained, and train the model；
E, using ultrasound wave irradiation distance h as preceding p binary systems of genes of individuals, using ultrasound wave irradiation time t as individual base
Rear q binary systems of cause, p and q are positive integer；PC host computer uses genetic algorithm, searches out best parameter group (h0, t0)；
F, PC host computer is according to best parameter group (h0, t0), transducer height regulating device is controlled by MCU controller,
Adjusting ultrasound wave irradiation distance is h0And the ultrasound wave irradiation time is controlled as t0Ultrasound wave irradiation is carried out, expected corresponding cell activity is obtained
Value.
Further, in step E, PC host computer by the transcription frequency of individual i at the decimal form of two parameters, in conjunction with
Corresponding impedance value Z is calculated in Z=f (h, t)i, pass through formula Ci=-(| Zi|-|Z0|)2Calculate impedance value fitness Ci, Zi
For the impedance value of individual i gene, Z0For the genetic test desired value of individual i.
It further, is 2 by ultrasound wave irradiation distance h points in step EpTime t of ultrasound wave irradiation point is 2 by a gradeqIt is a
Grade shares 2p+qKind may be individual, p < q.Each individual shared p+q binary coding, by duplication, intersection, makes a variation
To best parameter group (h0, t0), preceding p binary system is ultrasound wave irradiation distance h grade, and rear q binary system is the ultrasound wave irradiation time
T grade.
The present invention compared with technology, has the advantages that with existing method
1. the present invention can control the relevant parameter of ultrasonic irradiation by ultrasonic wave controller, then automatically regulate thin
The activity of born of the same parents.And existing cell activity adjusting method, it is all that artificial addition cell induction agent induces cell, artificial
Operation is more subjective, and will lead to the effect that cell activity is adjusted, there are errors, therefore the present invention can more accurately realize cell
Active Regulation.
2. the present invention obtains irradiation distance and irradiation time data corresponding with impedance value by test experiment, with BP nerve
Network fits the relationship of impedance value Z Yu irradiation distance h and irradiation time t, is then found offline most with genetic algorithm
Excellent combination finally carries out cell activity on-line control using apparatus of the present invention.In practical applications, the pass established according to hard measurement
It is model, it can be achieved that on-line control distance, time parameter can reach the requirement of cell activity adjusting, avoids complicated touch
Rope experiment.
3. cell activity adjusting method proposed by the present invention does not need to add in toward cell culture fluid compared to other methods
Add the apoptosis agent of various dose to induce cell apoptosis, but is induced by being shone with low-intensity low frequency ultrasound amplitude thin
Born of the same parents' apoptosis adjusts cell activity, to generate different grades of Activity Rank, reducing in the prior art will be in cell culture fluid
Impedance error caused by the cell death inducer of middle addition various dose various concentration.
4. the present invention by PC host computer directly with ultrasonic wave controller, controllor for step-by-step motor and electric cell-substrate impedance sensor
Output bus is connected, it can be achieved that cell activity, which adjusts experiment, tests co-ordination with cytoactive detection, has continuity, accurately
Property and high efficiency, suffer from broad application prospect in medicine, cytology and chemical field.
5. cell activity regulating device proposed by the present invention realizes that cell is living using the low-frequency ultrasonic irradiation of low-intensity
Property automatic adjustment, save and prepare induction agent and wait the long time that induces cell apoptosis, experiment is greatly saved
Time used also makes experiment become succinct easy to operate.
6, cell activity regulating device proposed by the present invention can prepare the sample of different cell activity grades, establish different
The cell activity of grade so that more accurate more easily carry out cytoactive detection experiment, therefore is determined in cell activity grade
In the case where, it can be used for the exploratory experiment of cytoactive detection method,
Detailed description of the invention
Fig. 1 is the overall structure diagram of the cell activity regulating device of Cell-oriented Activity determination of the present invention experiment；
Fig. 2 is the structural exploded view of electric cell-substrate impedance sensor 13 in Fig. 1；
Fig. 3 is the circuit control block diagram of Fig. 1.
The serial number and title of all parts in attached drawing: 1, cell incubator, 2, sliding rail, 3, sliding block, 4, screw rod, 5, experiment
Platform, 6, stepper motor, 7, step motor control line, 8, ultrasonic transducer controller, 9, PC host computer, 10, ultrasonic wave transducer
Device, 11, MCU controller, 12, electrode interface, 13, electric cell-substrate impedance sensor, 14, cell impedance detect controller, 16, input it is defeated
Bus out, 17, controllor for step-by-step motor, 18, culture dish, 19, array-type electrode, 20, culture dish pedestal.
Specific embodiment
Referring to Fig. 1, putting for cell activity regulating device of the present invention has cell incubator 1, peace in cell incubator 1
Fill ultrasonic transducer 10, electric cell-substrate impedance sensor 13 and transducer height regulating device.Ultrasonic transducer 10 is located at thin
The surface of born of the same parents' impedance transducer 13.Transducer height regulating device is fixedly connected with ultrasonic transducer 10, for adjusting ultrasound
Distance of the wave transducer 10 relative to electric cell-substrate impedance sensor 13.Ultrasonic transducer 10 connects ultrasonic transducer through control line
Controller 8, ultrasonic transducer controller 8 are mounted on the top of ultrasonic transducer 10, with ultrasonic transducer 10 simultaneously on
Lower adjusting.Electric cell-substrate impedance sensor 13 detects controller 14 through control line connection cell impedance, and cell impedance detects controller 14
Positioned at the lower section of electric cell-substrate impedance sensor 13.
It is PC host computer 9 and MCU controller 11 outside cell incubator 1.PC host computer 9 is connected by USB data line
MCU controller 11, MCU controller 11 connect the ultrasonic transducer inside cell incubator 1 through input/output bus 16 and control
Device 8, cell impedance detection controller 14 and transducer height regulating device.MCU controller 11 passes through ultrasonic transducer control
Device 8 processed can control ultrasonic transducer 10 and issue the ultrasonic wave of some strength I and fixed frequency f, and can control irradiation time t.
Transducer height regulating device is made of stepper motor 6, controllor for step-by-step motor 17, sliding block 3, screw rod 4.Stepping electricity
Machine 6 connects controllor for step-by-step motor 17 through step motor control line 7, and controllor for step-by-step motor 17 is connected through input/output bus 16
External MCU controller 11 is controlled the start and stop of stepper motor 6 by MCU controller 11 through controllor for step-by-step motor 17.Stepper motor
6 are arranged vertically in itself, and upward, the output shaft of stepper motor 6 is fixedly and coaxially connected the lower end of screw rod 4 to output shaft, match on screw rod 4
Trap has sliding block 3, and the side of sliding block 3 can be slided up and down along vertical sliding rail 2, and the side wall of sliding rail 2 is fixedly connected on cell culture
On the inner wall of case 1, the bottom of sliding rail 2 is fixedly connected with horizontal experimental bench 5.Another side wall of sliding block 3 extends to cell impedance biography
The surface of sensor 13 and it is fixedly connected with ultrasonic transducer 10 and ultrasonic transducer controller 8.When stepper motor 6 works
When, ultrasonic transducer 10 can be driven to move up and down by the cooperation of screw rod 4 and sliding block 3, with adjust ultrasonic transducer 10 to
Vertical range between the electric cell-substrate impedance sensor 13 of underface.
Referring to fig. 2, electric cell-substrate impedance sensor 13 is seated on experimental bench 5, and electric cell-substrate impedance sensor 13 is by array-type electrode
19 and culture dish 18 form, culture dish 18 is cylindrical glassware, in can hold cell culture fluid, ultrasonic transducer
10 can irradiate cell culture fluid.18 bottom of culture dish is culture dish pedestal 20, and culture dish pedestal 20 is seated in experimental bench 5
Corresponding recesses in.Tiled array-type electrode 19 on the inner bottom surface of culture dish 18, the pin electrode interface of array-type electrode 19
12 connect cell impedance from the inner wall of culture dish 18 after extraction detects controller 14.
Referring to Fig. 3, cell impedance detection controller 14 is made of DDS signal generator and signal picker.Cell impedance
The output end of input terminal of the array-type electrode 19 of sensor 13 through 12 connection signal collector of electrode interface, signal picker connects
MCU controller 11 is connect, signal picker can acquire the signal of electric cell-substrate impedance sensor 13 and amplify to signal, control convenient for MCU
The calculation processing of device 11 processed, MCU controller 11 will be in treated data transmissions to PC host computer 9.DDS signal generator it is defeated
Outlet connects electric cell-substrate impedance sensor 13, and the input terminal of DDS signal generator connects MCU controller 11, the control of MCU controller
DDS signal generator is used to generate the pumping signal of certain frequency and assignment.
Referring to Fig. 1-3, when the cell activity adjustment device work of Cell-oriented Activity determination of the present invention experiment, when cell hinders
When the energy for the ultrasonic irradiation that the cell in anti-sensor 13 receives ultrasonic transducer 10, mechanical damage can be caused to cell
Wound, a part of cell can be therefore dead, and another part cell will do it injury repair, to form different cell activity values.
The influence that the intensity I and irradiation time t of ultrasound wave irradiation adjust cell activity is maximum, and the frequency of ultrasound wave irradiation is to cell activity tune
The effect of section is minimum.Secondly, when ultrasonic wave is propagated in uniform dielectric, due to medium absorption and influence ultrasound intensity with distance
Increase and weaken, here it is acoustic attenuation.When propagating in air, ultrasonic wave initial intensity is I0, after h meters of distance,
Its intensity are as follows: Ih=I0e-2ah, a is absorption coefficient in formula.By the aerial propagation formula of ultrasonic wave it is found that ultrasound intensity I and
The relationship that propagation distance h exponentially successively decreases.Therefore it when needing certain ultrasound intensity I, can be converted into surpassing by PC host computer 9
The distance between acoustic wave transducer 10 and culture dish 18 h.It, then can be by adjusting one group of phase if necessary to a cell activity grade
The ultrasound parameter value (h, t) answered prepares the sample of required cell activity grade, and wherein ultrasound wave irradiation frequency is fixed as f (MHz).
Therefore, for cell activity regulating device of the present invention in experiment, first test experiment obtains cell impedance value Z and ultrasound wave irradiation distance h
With the correspondence sample data of irradiation time t, then with BP neural network algorithm sample instruction is carried out to collected sample data
Practice, obtain relation function Z=f (h, t), parameter off-line optimization is then carried out with genetic algorithm according to relation function Z=f (h, t),
Obtain the best parameter group (h of ultrasound wave irradiation distance h and irradiation time t0, t0), finally, according to best parameter group (h0, t0)
Carry out the desired cell activity value of on-line control.It is specific as follows:
Carry out test experiment first with cell activity regulating device of the present invention: MCU controller 11 is through controllor for step-by-step motor
17 control stepper motors 6 work, and so that ultrasonic transducer 10 is moved upwards up to highest point, this is ultrasonic transducer 10 original
Position, the maximum ultrasound wave irradiation distance between 13 bottom of ultrasonic transducer 10 and electric cell-substrate impedance sensor is h1.It is hindered in cell
Cell culture fluid is added dropwise in the culture dish 18 of anti-sensor 13, stands 2h.PC host computer 9 controls ultrasonic wave through MCU controller 11
Transducer controller 8, control ultrasound wave irradiation time are t1, ultrasonic transducer 10 irradiates cell culture fluid.It is irradiated
Afterwards, PC host computer 9 controls MCU controller 11 and detects 14 1 open signals of controller to cell impedance, at this point, cell impedance passes
Cell impedance value Z when the detection cell of sensor 13 is adherent11, and by cell impedance value Z11It is uploaded to PC host computer 9, PC host computer 9
It collects and records sample data (h1, t1, Z11).Then the cell culture in an electric cell-substrate impedance sensor 13 is replaced
New cell culture fluid identical in culture dish 18 is stood 2h, after PC host computer 9 controls 10 pairs of ultrasonic transducer replacements by liquid
Cell culture fluid irradiated, the control ultrasound wave irradiation time be t2, it is irradiated, it is detected by cell impedance detection controller 14
Cell impedance value Z12, PC host computer 9 collects and records down-sampled data (h1, t2, Z12).In this way, being protected in ultrasound wave irradiation distance
It holds as h1It under conditions of constant, moves in circles, until it is t that PC host computer 9, which obtains the ultrasound wave irradiation time,nCell impedance value Z1n,
It is sequentially t that PC host computer 9 obtains the ultrasound wave irradiation time altogether at this time1,t2,…,tnN sample data (h1, t1, Z11),
(h1, t2, Z12),…,(h1, tn, Z1n), first sampling period terminates.
Start to carry out second sampling period later: PC host computer 9 controls MCU controller 11 to controllor for step-by-step motor 17
One pulse signal, control stepper motor 6 rotate, and move down sliding block 3 to adjust ultrasonic transducer 10 and cell impedance and sense
The distance between 13 bottom of device obtains ultrasound wave irradiation distance h2.In ultrasound wave irradiation distance h2Under conditions of constant, using with first
A sampling period identical method, so that PC host computer 9 is obtained the ultrasound wave irradiation time is sequentially t1,t2,…,tnN sample
Data (h2, t1, Z21),(h2, t2, Z22),…,(h2, tn, Z2n), wherein Z21,Z22…,Z2nThe ultrasound wave irradiation time is sequentially respectively
t1,t2,…,tnWhen cell impedance value.
Again and so on, obtain the sample data (h in m-th of sampling periodm, t1, Zm1),(hm, t2, Zm2),…,
(hm, tn, Zmn), wherein hmUltrasound wave irradiation distance when being m-th of sampling period, Zm1,Zm2…,ZmnBe respectively ultrasound wave irradiation away from
From for hm, the ultrasound wave irradiation time be sequentially t1,t2,…,tnWhen cell impedance value.
So far entire data sample acquisition terminates, and acquires mn group data altogether.
After data sample acquires, PC host computer 9 is with ultrasound wave irradiation distance h (h1,h2,…,hm), irradiation time t (t1,
t2,…,tn) input as BP neural network, with cell impedance value Z (Z11,,…,Zmn) be trained as output, it obtains thin
Model Z=f (h, t) between born of the same parents' impedance value Z and ultrasound wave irradiation distance h and irradiation time t.For the convergence for accelerating training network
Property, PC host computer 9 is first to ultrasound wave irradiation distance h1、h2…hm, ultrasound wave irradiation time t1、t2…tnAnd cell impedance value Z11、
Z12…ZmnIt is normalized, respectively obtains corresponding ultrasound wave irradiation distance h '1、h’2…h’m, the sample ultrasound wave irradiation time
t’1、t’2…t’n, cell impedance value Z '11、Z’12…Z’mn, data sample after obtained normalized is then divided into training
Sample and test sample, training sample is for creating BP neural network model, and test sample is for correcting the model.To train number
According to the input variable of ultrasound wave irradiation distance h, ultrasound wave irradiation time t as BP neural network in sample, with cell impedance value Z work
For output variable, reasonable BP neural network model is established.Test amendment is carried out to the model with test data sample, by repairing
The just Parameters in Mathematical Model is finally completed so that the relative error between actual measured value and model output value is small as far as possible
The training process of entire BP neural network.The mathematics obtained between cell impedance value Z and ultrasound wave irradiation distance h, irradiation frequency t closes
It is formula Z=f (h, t).
After the completion of BP neural network training, PC host computer 9 is to ultrasound wave irradiation distance h and irradiation time t the two parameters
Best parameter group (h is found using genetic algorithm0, t0), it is operated by duplication, intersection, variation etc., is phased out fitness
The low solution of functional value increases the high solution of fitness function value, the optimal solution that evolution N is instead of needed afterwards, to be wanted
Cell activity value.Offline parameter optimizing is divided into classification and the coding, genetic algorithm off-line optimization of parameter, specific as follows:
1. classification and the coding of parameter: by the ultrasound wave irradiation between 13 bottom of ultrasonic transducer 10 and electric cell-substrate impedance sensor
Distance h points are 2pA grade (p is positive integer), as preceding p binary systems of genes of individuals, MCU controller 11 gives stepper motor
17 1 pulse signals of controller, control stepper motor 6 rotate, and control to adjust hr=hm+ a × r, r=0,1,2 ..., 2p, formula
Middle hrDistance when being grade r, hmIt is the minimum range of ultrasonic transducer 10 Yu electric cell-substrate impedance sensor 13, h1It is ultrasonic transduction
Maximum distance between device 10 and electric cell-substrate impedance sensor 13, a are the difference of neighbor distance grade, i.e. a=(h1-hm)/8。
It is 2 by time t of ultrasound wave irradiation pointqA grade (q is positive integer) is led to as rear q binary systems of genes of individuals
MCU controller 11 is crossed to 8 one timer signals of ultrasonic transducer controller, controls the time t of ultrasound wave irradiation, i.e. tk=t1+g
× k, k=0,1,2 ..., 2q, t in formulakIrradiation time when for grade being k, t1For the initial time of ultrasound wave irradiation, when g is adjacent
Between grade time difference.
The two parameter combinations one of ultrasound wave irradiation distance h, irradiation time t share 2 as a result,p+qKind may individual, wherein p <
q.Each individual shared p+q binary coding, preceding p binary systems are the grade of ultrasound wave irradiation distance h, control from left to right
Ultrasound wave irradiation distance；Afterwards q be ultrasound wave irradiation time t grade, control the time of ultrasound wave irradiation.
2. genetic algorithm off-line optimization.Using genetic algorithm, obtains the device and adjust the optimal of two parameters of cell activity
Combination, specific work step are as follows:
Step 1: expected cell impedance value Z is arranged in PC host computer 90, evolutionary generation d=0 is initialized, maximum evolve is set
Algebra dmax, setting crossover probability is Pc, setting mutation probability is Pm。
Step 2: PC host computer 9 from 2p+q2 are randomly choosed during kind is possiblepIndividual composition group T0, and it is compiled
Number, from individual 1 to individual 2p。
Step 3: PC host computer 9 initializes individual count parameter i=1.
Step 4: PC host computer 9 by the transcription frequency of individual i at the decimal form of two parameters, and marriage relation letter
Number Z=f (h, t) is calculated, and impedance value Z is obtainedi。
Step 5: PC host computer 9 calculates and records impedance value fitness Ci, fitness function is as follows:
Ci=-(| Zi|-|Z0|)2
Wherein ZiFor the detection impedance value of individual i gene, Z0For the genetic test desired value of individual i, CiIt is bigger, impedance value
More meet the requirements.
Step 6: PC host computer 9 executes i=i+1.
Step 7: judge parameter i > 2pIt is whether true, if invalid, in group 2pIndividual has asked suitable not yet
Response needs to jump to step 4；If i > 2pIt sets up, then 2pThe fitness value of individual has all been asked, and following step is executed
Eight.
Step 8: termination condition judgement.Evolutionary generation d is added 1 by PC host computer 9, then judges d > dmaxIt is whether true, such as
Fruit is invalid, executes following step nine；By the individual with maximum adaptation degree obtained in evolutionary process if setting up
Gene is exported as optimal solution, executes following step ten three.
Step 9: PC host computer 9 carries out duplication operation, selectes 1 individual at random from group T every time, and dyed
Body duplication, the selected probability of each individual is Pi, repeat 2pIt is secondary, obtain a population T1。PiIt is that the fitness of each individual accounts for
The ratio of total fitness, it is known that group size 2p, the fitness value of individual i is Ci, then individual i selected probability PiAre as follows:
Step 10: PC host computer 9 carries out crossover operation, by crossing-over rate PcThe chromosome number c of the intersection determined, from group
T1In determine that c chromosome carries out crossover operation at random: obtain 2 will be selected before crossover operationpIndividual group in a random way
At 2p-1It, should be by P when carrying out intersection to pairing group of individualscTwo individual part-structures behind the crosspoint determined carry out mutual
It changes, generates two new individuals, repeat 2p-1It is secondary, original chromosome is replaced with the new chromosome of generation, obtains group T2。
Step 11: PC host computer 9 carries out mutation operation, by aberration rate PmThe variation number m determined, from T2In it is random
It determines m chromosome, carries out mutation operation respectively, replace original chromosome with the new chromosome of generation, obtain group T3。
Step 12: by group T3As population of new generation, that is, use T3Instead of T0, go to step 3.
Step 13: the genes of individuals of output is divided into two sections of extractions by PC host computer 9, obtains optimal combination, the preceding position p two
System is the ultrasound wave irradiation distance h grade of optimum combination；Q binary systems are the ultrasound wave irradiation time t grade of optimum combination afterwards, and
By two parameter (h of optimum combination0, t0) be stored in PC host computer 9 with decimal form.
Finally, according to best parameter group (h0, t0) carrying out the desired cell activity value of on-line control: PC host computer 9 is according to something lost
Best parameter group (the h that propagation algorithm parameter off-line optimization obtains0, t0), stepper motor 6 is controlled by MCU controller 11 and is rotated,
Adjusting ultrasound wave irradiation distance is h0, and the ultrasound wave irradiation time is controlled as t0, ultrasound wave irradiation is carried out, it is living to obtain expected corresponding cell
Property value.Since there are certain mathematical relationship, i.e. Q=g (Z), to can be expected with cell impedance value for cell activity value
Cell activity value.
Claims (7)
1. a kind of cell activity regulating device of Cell-oriented Activity determination experiment, including electric cell-substrate impedance sensor, it is characterized in that:
It is ultrasonic transducer right above electric cell-substrate impedance sensor, it is opposite that ultrasonic transducer is fixedly connected with adjusting ultrasonic transducer
The transducer height regulating device of distance between electric cell-substrate impedance sensor；Electric cell-substrate impedance sensor is through control line connection cell resistance
Anti- detection controller, ultrasonic transducer connect ultrasonic transducer controller through control line；PC host computer is connected by data line
MCU controller is connect, MCU controller is separately connected ultrasonic transducer controller, cell impedance detection controller and energy converter
Height adjustment device.
2. the cell activity regulating device of Cell-oriented Activity determination experiment according to claim 1, it is characterized in that: described
Transducer height regulating device be made of stepper motor, controllor for step-by-step motor, sliding block and screw rod, stepper motor is through control line
Controllor for step-by-step motor is connected, controllor for step-by-step motor connects MCU controller；Stepper motor is arranged vertically, and output shaft is coaxial
It is fixedly connected with the lower end of screw rod, cooperation is cased with sliding block on screw rod, and sliding block is fixedly connected with ultrasonic transducer.
3. the cell activity regulating device of Cell-oriented Activity determination experiment according to claim 1, it is characterized in that: cell
Impedance transducer is made of array-type electrode and culture dish, is contained with cell culture fluid in culture dish, on the inner bottom surface of culture dish
Tiling has array-type electrode, and array-type electrode detects controller through electrode interface connection cell impedance.
4. the cell activity regulating device of Cell-oriented Activity determination experiment according to claim 3, it is characterized in that: cell
Impedance detection controller is made of DDS signal generator and signal picker, the input of array-type electrode connection signal collector
End, the output end of signal picker connect MCU controller, and the output end of DDS signal generator connects electric cell-substrate impedance sensor,
The input terminal of DDS signal generator connects MCU controller.
5. a kind of cell activity adjusting method of cell activity regulating device as described in claim 1, it is characterized in that include with
Lower step:
A, PC host computer controls transducer height adjustment device work through MCU controller, is moved upwards up to ultrasonic transducer
The home position of highest point, ultrasound wave irradiation distance is h at this time1；Cell culture fluid is added dropwise in electric cell-substrate impedance sensor, PC is upper
The ultrasound wave irradiation time that machine controls ultrasonic transducer is t1, electric cell-substrate impedance sensor detects cell impedance value Z11；Then more
Cell culture fluid is changed, the control ultrasound wave irradiation time is t2, detect cell impedance value Z12；It loops back and forth like this, until PC upper
The machine acquisition ultrasound wave irradiation time is sequentially t1,t2,…,tnN sample data (h1, t1, Z11),(h1, t2, Z12),…,
(h1, tn, Z1n), first sampling period terminates；
B, PC PC control ultrasonic transducer moves down, and obtaining ultrasound wave irradiation distance is h2, in ultrasound wave irradiation distance h2Constant
Under the conditions of, the PC host computer acquisition ultrasound wave irradiation time is sequentially t1,t2,…,tnN sample data (h2, t1, Z21),
(h2, t2, Z22),…,(h2, tn, Z2n), second sampling period terminates；
C, and so on, obtain the sample data (h in m-th of sampling periodm, t1, Zm1),(hm, t2, Zm2),…,(hm, tn,
Zmn), total mn group data；
D, PC host computer is with ultrasound wave irradiation distance h1,h2,…,hm, irradiation time t1,t2,…,tnAs the defeated of BP neural network
Enter, with cell impedance value Z11,…,ZmnAs output, BP neural network model Z=f (h, t) is obtained, and train the model；
E, using ultrasound wave irradiation distance h as preceding p binary systems of genes of individuals, using ultrasound wave irradiation time t as genes of individuals
Q binary systems afterwards, p and q are positive integer；PC host computer uses genetic algorithm, searches out best parameter group (h0, t0)；
F, PC host computer is according to best parameter group (h0, t0), transducer height regulating device is controlled, ultrasound wave irradiation distance is adjusted
For h0And the ultrasound wave irradiation time is controlled as t0Ultrasound wave irradiation is carried out, is obtained mutually it is contemplated that cell activity value.
6. the cell activity adjusting method of cell activity regulating device according to claim 5, it is characterized in that: in step E,
At the decimal form of two parameters, corresponding resistance is calculated in conjunction with Z=f (h, t) in the transcription frequency of individual i by PC host computer
Anti- value Zi, pass through formula Ci=-(| Zi|-|Z0|)2Calculate impedance value fitness Ci, ZiFor the impedance value of individual i gene, Z0It is a
The genetic test desired value of body i.
7. the cell activity adjusting method of cell activity regulating device according to claim 5, it is characterized in that: in step E,
It is 2 by ultrasound wave irradiation distance h pointspTime t of ultrasound wave irradiation point is 2 by a gradeqA grade, shares 2p+qKind may be individual, and p <
Q, each individual shared p+q binary coding, obtains best parameter group (h by duplication, intersection, variation0, t0), preceding p
Position binary system is ultrasound wave irradiation distance h grade, and rear q binary system is ultrasound wave irradiation time t grade.
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