CN106754357B - Wireless program-controlled illumination system and application for light science of heredity cell high pass quantity research - Google Patents
Wireless program-controlled illumination system and application for light science of heredity cell high pass quantity research Download PDFInfo
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- CN106754357B CN106754357B CN201611095124.8A CN201611095124A CN106754357B CN 106754357 B CN106754357 B CN 106754357B CN 201611095124 A CN201611095124 A CN 201611095124A CN 106754357 B CN106754357 B CN 106754357B
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/06—Means for regulation, monitoring, measurement or control, e.g. flow regulation of illumination
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Abstract
The invention discloses a kind of wireless program-controlled illumination system for light science of heredity cell high pass quantity research, which includes host computer and control software, wireless communication module, slave computer central processing unit, slave computer register die group, slave computer light emitting diode group and constant voltage dc source.The device of the invention studies the research of Drug molecule screening especially relevant to signal path for the light science of heredity signal path of external scale evaluation cell, it solves the plant issue for lacking high-throughput, more intensity, long-time, the stimulated in vitro of multifactor illumination in presently relevant research, effectively increases the accuracy and conventional efficient of light stimulus system in light genetics research.In addition, the condition based on this system culture cell photoactivation obtained, such as intensity of illumination, photoactivation time also have wide practical use for the light genetics research of cell in future.
Description
Technical field
The invention belongs to integrated electronic circuits and light genetic arts, in particular to a kind of to be used for light science of heredity cell high pass
The wireless program-controlled illumination system of quantity research and application.
Background technique
Light genetics technology is that one rapidly developed in recent years incorporates the complete of optics, genetic engineering and electronic engineering
The biotechnology of new multi-crossed disciplines.Using photogene technical controlling cellular activity, there is not damaged, non-intruding, time-space resolution
Rate is high, can quantitatively repeat, using the advantages such as simple, is got in research fields such as Neuscience, cell biology, signal paths
To be more widely applied.
So far, by fiber-coupled diode fixed point local stimulation neuron (such as patent CN201510654472.3)
Or utilize Minitype LED array multiple spot light stimulus (such as Kwon K, Li W.Integrated multi-LED array with
three-dimensional polymer waveguide for optogenetics[C]//Micro Electro
Mechanical Systems(MEMS),2013IEEE 26th International Conference on.IEEE,2013:
1017-1020.), light science of heredity obtains very big achievement in neuroscience field.
But in more emerging signal path research field, light stimulus system also very it is simple and crude (such as Bugaj L J,
Choksi A T,Mesuda C K,et al.Optogenetic protein clustering and signaling
Activation in mammalian cells [J] .Nature methods, 2013,10 (3): 249-252.): 1. lamp bead number
Amount is few, not can be carried out high-throughput signal path molecule screening；2. can only be with single brightness irradiated stimulator cell, it cannot be from software
On fine-tune brightness with probe into intensity of illumination and signal path activation degree relationship；3. passing through physical connection and computer expert
Letter is not easy under incubator environment irradiate culture studies for a long time to cell.4. the same time can only be pierced with a kind of light
Swash.However signal path is studied, Drug molecule screening especially relevant to signal path need high-throughput, more intensity, it is long when
Between and/or multifactor incentive condition.
Summary of the invention
In view of it is above-mentioned in the prior art the shortcomings that, the present invention provides a kind of for the wireless of light science of heredity high pass quantity research
Program-controlled illumination system, the system can be improved light genetics technology conventional efficient, reduce experimental cost, which can be applied to
Carry out the control of light science of heredity high throughput in the cell of in vitro culture.
A kind of wireless program-controlled illumination system for light science of heredity high pass quantity research, comprising: host computer PC, bluetooth channel radio
Interrogating module, slave computer and constant voltage dc source, the slave computer includes: the centre positioned at IC printed board one side
Manage unit, register die group, communication interface and power interface；Positioned at the light emitting diode of IC printed board another side
Group；And light emitting diode distribution is corresponding with each hole site of orifice plate used in biological cell assays；
Control instruction is delivered to centre by blue tooth wireless communication module by the control software in the host computer PC
Manage unit；
The central processing unit receives the control instruction that blue tooth wireless communication slave is sent, and by control instruction and when
Clock signal input register chipset；
The serial input of control instruction is converted to parallel output by the register die group, and controls light emitting diode
Light-emitting mode, individually control each Light-Emitting Diode shine；
The light emitting diode group implements light stimulus to cell in light genetics research；
The constant voltage dc source is used to provide work constant voltage dc source to the component in slave computer.
Control software in the host computer PC is for setting light emitting diode (LED) bright light duration, flashing mode, light
Brightness and photochromic.
The blue tooth wireless communication module includes: blue tooth wireless communication host and blue tooth wireless communication slave, bluetooth without
Line host computer communication is connected by USB interface with host computer, and blue tooth wireless communication slave is assisted with blue tooth wireless communication host with bluetooth
View pairing；The blue tooth wireless communication host is instructed for emission control, and the blue tooth wireless communication slave is for receiving control
Instruction.
Preferably, the central processing unit, register die group and light emitting diode group welding are integrated at one piece
On circuit board.
Preferably, the register die group includes processor register die group and light emitting diode register die
Group, processor register die group are connected with light emitting diode register die group, convert control luminous two for control signal
The level signal of the specific on-off of pole pipe.
Preferably, the light emitting diode group includes 96 light emitting diodes, at 8 × 12 rectangular arrangeds, each luminous two
Centre distance is 9mm between pole pipe, corresponding with standard each hole site of 96 porocyte culture plates used in biological cell assays.
Preferably, the LED operation frequency is 40~110Hz, adjusts operating brightness with PWM mode.Due to
The biological cell of in vitro culture is the insensitive body cell of frequency rather than the neuronal cell sensitive to light frequency, therefore works as illumination
After frequency is greater than 40Hz, illumination frequency ignores to impact cell.Further preferred illumination frequency is 50Hz, improves control essence
Degree.
Preferably, the light emitting diode outgoing light wavelength include 470nm blue light, 520nm green light, 620nm feux rouges, into
One step is preferred, and light emitting diode outgoing light wavelength is 470nm.
Light genetics research relies on light-dependent control element (photosensitive protein) and is transferred to target cell, and the activation of different light-dependent control elements needs
The coloured light of different wave length stimulates.Optimal wavelength is the blue light of 470nm, can effective stimulus cryptochrome class ligth activated element
CRY2/CIBN becomes to closing: after cryptochrome class photosensitive protein CRY2 absorbs 470nm blue light, conformation changes, thus and ligand
PROTEIN C IBN is combined；And after losing blue light illumination, cryptochrome class photosensitive protein CRY2 whithin a period of time with ligandin
CIBN separation, realizes the light-operated reversible of light heredity light-dependent control element.Therefore wavelength is the blue light stimulus of 470nm by preferably
Applied in cell-signaling pathways research.
The physical size of the IC printed board is matched with 96 porocyte culture plates of standard.
The slave computer is connected by data flat cable with the blue tooth wireless communication slave.
The DC power supply is connect by power interface with the slave computer.
The course of work of wireless program-controlled illumination system: the control software in host computer issues control instruction to bluetooth channel radio
Host is interrogated, control instruction is transferred to blue tooth wireless communication slave, bluetooth by blue tooth wireless communication host in a manner of wireless blue tooth
Control instruction is delivered to the central processing unit by communication interface by wireless telecommunications slave, and central processing unit refers to control
Order and clock signal input register die group, register die group controls the light-emitting mode of light emitting diode, and it is right to control its
Cell in light genetics research implements light stimulus, completes entire illumination.
Wireless program-controlled illumination system of the invention is real for carrying out the control of light science of heredity high throughput in the cell of in vitro culture
The wireless multithreading optics regulation under the conditions of cells in vitro i.e. incubator is showed, and setting is preferably used for CIBN/CRY2 system
The incentive condition of activation.
In the present invention, the light stimulus of single-chip microcontroller and integrated circuit technique exploitation light genetics research field cell experiment is utilized
System improves the accuracy and reliability of system, compared with the existing light genetics technology studied applied to signal path, this
Inventing has technical effect beneficial below:
(1) instrument utilization rate is high, and 96 light emitting diodes and 96 porocyte culture plates that illumination system has cooperate, each
LED lamp bead can individually be controlled by register die, thus single experiment can carry out repeatedly it is non-interfering quantitative
Analysis；
(2) light stimulus regulation is accurate, and the light intensity of light emitting diode can be by software adjustment linear change, the sudden strain of a muscle of light emitting diode
Bright frequency stabilization, is not present error；
(3) system both can be irradiated stimulation to cell in normal culture environment, can also pass through wireless control
Culture is irradiated in realization for a long time in incubator environment to cell；
(4) a variety of monochrometric stimulus can import a variety of light-operated albumen, realize the research to act synergistically to signal path；
(5) apparatus structure is succinct, easy to use, is quickly adjusted conducive to according to experiment condition.
Detailed description of the invention
Fig. 1 is overall workflow frame diagram of the present invention；
Fig. 2 is the next machine face (light emitting diode face) structure chart in the embodiment of the present invention；
Fig. 3 is slave computer reverse side in the embodiment of the present invention (component face) structure chart；
Fig. 4 is the illumination operation schematic diagram in the embodiment of the present invention to 96 orifice plates.
Specific embodiment
In order to more specifically describe the present invention, with reference to the accompanying drawing and specific embodiment is to technical solution of the present invention
It is described in detail.
As shown in Figure 1, the present invention is used for the wireless program-controlled illumination system of light science of heredity high pass quantity research, comprising: host computer
1, Bluetooth wireless communication module 2, slave computer central processing unit 3, slave computer register die group 4 and slave computer LED group 5.
Host computer 1 is the interactive program operated on PC, is the operation that user's control adjusts each LED light stimulus modality
Interface, brightness (PWM duty cycle), bright light duration, flashing mode and photochromic four parameters of photic stimuli are adjusted.Bluetooth
Wireless communication module 2 includes Bluetooth wireless communication host and Bluetooth wireless communication slave, and host turns USB interface by serial ports and realizes
With the communication of host computer 1, slave is then connected with slave computer data-interface 7, and after host is matched with slave, host is by host computer
1 control instruction data packet is sent to slave with Bluetooth signal, is sent to slave computer central processing unit by slave computer data-interface 7
3.Slave computer central processing unit 3 is by control instruction and clock signal transmission to slave computer register die group 4.Slave computer register
Chipset 4 realizes the on-off control of slave computer LED group 5 by the cathode level height of control slave computer LED group 5, and then controls
The brightness of slave computer LED group 5, bright light duration, flashing mode with it is photochromic.
As shown in Figures 2 and 3, the present invention is for the bottom in the wireless program-controlled illumination system of light science of heredity high pass quantity research
Machine includes: slave computer LED group 5, slave computer IC printed board 6, slave computer central processing unit 3, the deposit of slave computer processor
Device chipset 41, slave computer LED register die group 42, slave computer data-interface 7, slave computer power interface 8.Wherein, the next
Machine data-interface 7 is connected with slave computer central processing unit 3, and PC control signal is passed to slave computer central processing unit 3；It is the next
Machine central processing unit 3 is connected with slave computer processor register die group 41, for extending the I/O of slave computer central processing unit 3
Port, transmitting control signal；Slave computer processor register die group 41 is connected with slave computer LED register die group 42,
Convert control signal to the level signal of the control specific on-off of LED；Slave computer LED group 5 includes 96 LED, is in 8 × 12 rows
Column, each adjacent LED distance are 9mm to match distance between each adjacent holes of 96 orifice plates；External dc power passes through the next electromechanical source
Interface 8 provides power supply for above-mentioned all slave computer components, and has preferable filter stabilization.
The program of host computer 1 is write using C#；Bluetooth wireless communication module is using HC-06 and USB-TTL module；Slave computer
Data-interface 7 is using row's needle socket；Slave computer central processing unit 3 uses STC15W408AS；Slave computer processor register die
Group 41 uses 74HC595；Slave computer LED register die group 42 uses PIC10F200 for core circuit；Slave computer LED group 5 is adopted
With common anode three-color LED；Slave computer power interface 8 uses DC005 power outlet and filter regulator circuit；Power supply using AC220V~
DC5V 10A pressure stabilizing adapter.The fixed 50Hz of the working frequency of LED；Brightness, brightness degree 100 are adjusted with PWM mode；Feux rouges work
Making wavelength is 630~640nm；Green light operation wavelength is 515-525nm；Blue light operation wavelength is 465-475nm；Cell trench bottom
Three kinds of light highlight fluxes are 90lx.
Illumination work structuring such as Fig. 4 institute of the present invention for the wireless program-controlled illumination system of light science of heredity high pass quantity research
Show, wherein special 96 orifice plate 12 transparent 96 orifice plate common different from market, but 96 orifice plate of black non transparent, to avoid one
The LED group 5 of a cell cultivation groove 11 impacts another culture tank.96 orifice plate plate lids 10 be also it is special, it is normal with market
See plate lid it is identical be that capping is transparent, fit 96 orifice plates, but more black non transparent card slots 9 right above each cell cultivation groove,
To support light control system slave computer IC printed board 6 and each LED light source 5 be isolated.Particularly, to avoid transparency cover 10
In diffusing reflection, 9 base portion of black non transparent card slot extends to 10 bottom of plate lid.
Workflow are as follows: cell culture fluid is injected into 96 orifice plate cell slots 11；The lid of plate lid 10 is put on 96 orifice plates 12；It will
Downwards, each LED is aligned with plate lid card slot 9 and is buckled on plate lid 10 light control system slave computer LED；By 96 orifice plates 12 and light control system
Slave computer is put into togerther cell incubator；Light control system host computer PC 1 sets illumination parameter and sends；Light control system slave computer is opened
Beginning light stimulation,；After predetermined time arrives, host computer PC client 1 issues early warning, and slave computer stops illumination.
Compared to the method for filling reagent using multiple culture dishes and gradation in traditional illumination research, the present invention has cooperated carefully
Common 96 well culture plate in born of the same parents' experiment, less reagent can be used disposably to be filled by the volley of rifle fire, therefore use nothing of the invention
Thread control illumination system is greatly improved light genetics technology conventional efficient, while experimental cost is effectively reduced, and has reached high pass
Measure the purpose of optically screening.
Technical solution of the present invention and beneficial effect is described in detail in above-described specific embodiment, Ying Li
Solution is not intended to restrict the invention the foregoing is merely presently most preferred embodiment of the invention, all in principle model of the invention
Interior done any modification, supplementary, and equivalent replacement etc. are enclosed, should all be included in the protection scope of the present invention.
Claims (5)
1. a kind of wireless program-controlled illumination system for light science of heredity high pass quantity research, comprising: host computer PC, blue tooth wireless communication
Module, slave computer and constant voltage dc source, it is characterised in that: the slave computer includes: positioned at IC printed board one
Central processing unit, register die group, communication interface and the power interface in face；Positioned at IC printed board another side
Light emitting diode group；The light emitting diode group includes 96 light emitting diodes, at 8 × 12 rectangular arrangeds, each light-emitting diodes
Centre distance is 9mm between pipe, corresponding with standard each hole site of 96 porocyte culture plates used in biological cell assays；
Control instruction is delivered to central processing list by blue tooth wireless communication module by the control software in the host computer PC
Member；
The blue tooth wireless communication module includes: blue tooth wireless communication host and blue tooth wireless communication slave, bluetooth channel radio
News host is connected by USB interface with host computer PC, and blue tooth wireless communication slave and blue tooth wireless communication host are with Bluetooth protocol
Pairing；
The central processing unit receives the control instruction that blue tooth wireless communication slave is sent, and control instruction and clock are believed
Number input register chipset；
The serial input of control instruction is converted to parallel output by the register die group, and controls the bright of light emitting diode
Lamp duration, flashing mode, lamplight brightness and photochromic individually control shining for each light emitting diode；
The outgoing light wavelength of the light emitting diode includes 470nm, 520nm and 620nm, used light-emitting diodes plumber
Working frequency is 40 ~ 100Hz, adjusts operating brightness with PWM mode, implements light stimulus to cell in light genetics research；
The constant voltage dc source is used to provide work constant voltage dc source to the component in slave computer.
2. wireless program-controlled illumination system according to claim 1, it is characterised in that: the object of the IC printed board
It is corresponding with 96 porocyte culture plates of standard to manage size.
3. wireless program-controlled illumination system according to claim 1, it is characterised in that: the register die group includes place
Manage device register die group and light emitting diode register die group, processor register die group and light emitting diode register
Chipset is connected, and converts control signal to the level signal of the control specific on-off of light emitting diode.
4. wireless program-controlled illumination system according to claim 1, it is characterised in that: the central processing unit, deposit
Device chipset and light emitting diode group welding are on one piece of integrated circuit board.
5. carrying out light heredity in the cell that described in any item wireless program-controlled illumination systems are cultivated in vitro according to claim 1 ~ 4
Learn the application in high-throughput control.
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| CN201611095124.8A CN106754357B (en) | 2016-12-01 | 2016-12-01 | Wireless program-controlled illumination system and application for light science of heredity cell high pass quantity research |
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| CN106754357B true CN106754357B (en) | 2019-06-11 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111103272B (en) * | 2018-10-26 | 2022-08-05 | 山东大学 | Real-time screening and measuring system and method for cell specific photosensitive effect |
| CN109735450A (en) * | 2019-03-11 | 2019-05-10 | 中国医学科学院生物医学工程研究所 | A kind of low energy light source shelf for cell incubator |
| WO2022126363A1 (en) * | 2020-12-15 | 2022-06-23 | 深圳先进技术研究院 | Cell phenotype control device |
| CN112831418B (en) * | 2021-04-07 | 2024-04-23 | 广州市丹树生物科技有限责任公司 | In-vitro high-throughput screening device for phototherapy and application thereof |
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| EP3092474B1 (en) * | 2014-06-12 | 2020-04-22 | Axion Biosystems, Inc. | Multiwell microelectrode array with optical stimulation |
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| TW201229235A (en) * | 2011-01-12 | 2012-07-16 | Hsiuping Inst Technology | Universal optogenetic cell experimental system |
| WO2013056037A1 (en) * | 2011-10-13 | 2013-04-18 | The Cleveland Clinic Foundation | Estimation of neural response for optical stimulation |
| CN203287381U (en) * | 2013-06-05 | 2013-11-13 | 青岛市中心血站 | 96-well plate wrong-sample-adding-resistant device |
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