CN107328955A - Cellular electrophysiologicalsensor detection method, the device and system of scanning probe are excited based on light - Google Patents
Cellular electrophysiologicalsensor detection method, the device and system of scanning probe are excited based on light Download PDFInfo
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- CN107328955A CN107328955A CN201710495757.6A CN201710495757A CN107328955A CN 107328955 A CN107328955 A CN 107328955A CN 201710495757 A CN201710495757 A CN 201710495757A CN 107328955 A CN107328955 A CN 107328955A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01Q—SCANNING-PROBE TECHNIQUES OR APPARATUS; APPLICATIONS OF SCANNING-PROBE TECHNIQUES, e.g. SCANNING PROBE MICROSCOPY [SPM]
- G01Q60/00—Particular types of SPM [Scanning Probe Microscopy] or microscopes; Essential components thereof
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Abstract
The scanning cellular electrophysiologicalsensor detection method of probe, device and system are excited based on light the invention discloses a kind of.This method includes：The conductive pinpoint of driving scanning probe is contacted with the cell membrane sample on sample substrate, and the cell membrane sample is excited using LASER Light Source so that the cell membrane sample obtains photoelectric characteristic response；By the conductive pinpoint to the cell membrane sample extrinsic motivated electric signal；Using ion channel pattern and photoelectric respone electric signal of the cell membrane sample when light excites lower and extrinsic motivated electric signal described in the scanning probe records.The present invention can detect ion channel in cell membrane photoelectric characteristic and ion channel nanoscale three-dimensional appearance simultaneously, and not destroy sample structure and property.
Description
Technical field
The thin of probe is scanned the present embodiments relate to biological electricity detection technology field, more particularly to a kind of excited based on light
Born of the same parents' electro physiology detection method, device and system.
Background technology
Biological electricity is played a significant role in plant growth vital movement, and electric signal all can to many physiological activities of plant
Produce material impact, iuntercellular passes through one of basic mode that electric signal conducts information is that plant information is conducted.Scientist is in life
Change of the cell biological film to ion permeability is found that in the research of thing electricity generation mechanism.The spy that ion channel is produced by cell
Different protein is constituted, and they gather together and are embedded on cell membrane.The activity of ion channel is to realizing that cell various functions have
It is significant.The function of ion channel on cell membrane, the osmotic pressure except that can adjust intraor extracellular is also to maintain cell membrane
The important molecule of current potential, and cell will carry out electric signal conduction, be the turnover by ion to cause the change of film potential.Laser
Effect can significantly affect the activation of opsin photo-sensitive ion passage stable state and inactivation.Dim light acting photosensitive ion channel cell membrane
Voltage gated ion channel characteristic can be changed, so as to influence the formation and granting of action potential, regulation of physiological functions.It is right at present
The detection method of photo-sensitive ion passage still lacks, and cell membrane light can be obtained simultaneously and excites bioelectrical signals and epicyte protein high-precision
The technological means for spending structure is still not enough.
The method of tradition research cell membrane electrical characteristics and ion channel performance includes：Patch clamp technique, channel protein separation
Biochemical technology, artificial membrane ion channel reconstitution technology and some physics and the chemical technologies such as purifying.Can be with using patch clamp technique
Photosensitive ion channel or photopigment modulation of ion channels electric current and its opening and closing time-histories are directly observed and differentiated, but can not be visited in real time
Survey cell membrane nanostructured morphologies, it is impossible to while obtaining the structural information and electrical characteristics of cell membrane nanoscale.
The content of the invention
Present invention offer is a kind of to excite the cellular electrophysiologicalsensor detection method for scanning probe, device and system based on light, with reality
While present sample surfaces detection nanoscale pattern, ion channel in cell membrane light is obtained in real time and excites electric signal.
In a first aspect, the embodiments of the invention provide it is a kind of based on light excite scanning probe cellular electrophysiologicalsensor detection side
Method, this method includes：
The conductive pinpoint of driving scanning probe is contacted with the cell membrane sample on sample substrate, using LASER Light Source
Excite the cell membrane sample so that the cell membrane sample obtains photoelectric characteristic response；
By the conductive pinpoint to the cell membrane sample extrinsic motivated electric signal；
Excited and led to the ion under extrinsic motivated electric signal in light using cell membrane sample described in the scanning probe records
Road pattern and photoelectric respone electric signal.
Preferably, using it is described scanning probe records described in cell membrane sample light excite with extrinsic motivated electric signal
Ion channel pattern and electric signal are specifically included：The cell membrane sample is detected using the scanning probe scanning and excites bar in light
Ion channel pattern under part, ion channel of the cell membrane sample under photo-excitation conditions is detected using the scanning probe
Photoelectric respone electric signal；
Excited and led to the ion under extrinsic motivated electric signal in light using cell membrane sample described in the scanning probe records
After road pattern and photoelectric respone electric signal, in addition to：According to the ion channel shape of ion channel morphogenesis cell membrane sample
Looks figure, according to the photoelectric respone electric signal distribution map of the photoelectric respone electric signal cellulation membrane sample of each ion channel.
Preferably, the cell membrane sample is excited to specifically include using LASER Light Source：Regulate and control the optical maser wavelength of LASER Light Source
And intensity, using cell membrane sample described in the laser excitation of different wave length and varying strength；Using the scanning probe records institute
State cell membrane sample and excited in light and specifically included with the ion channel pattern and photoelectric respone electric signal under extrinsic motivated electric signal：
Using ion channel of the cell membrane sample under the laser excitation of different wave length and varying strength described in the scanning probe records
Pattern and photoelectric respone electric signal.
Preferably, using it is described scanning probe records described in cell membrane sample light excite with extrinsic motivated electric signal
After ion channel pattern and photoelectric respone electric signal, in addition to：
According to the ion channel pattern and photoelectric respone electric signal of the cell membrane sample, the conductive pinpoint positioning is driven
Pinpointed in the ion channel of cell membrane sample, change the wavelength and/or intensity of the LASER Light Source, it is thin using scanning probe records
The local photoelectric respone electric signal that the ion channel of after birth sample is pinpointed under the laser excitation of different wavelength and/or intensity becomes
Change.
Preferably, using it is described scanning probe records described in cell membrane sample light excite with extrinsic motivated electric signal
After ion channel pattern and photoelectric respone electric signal, in addition to：
Change cell membrane sample environment, the photoelectric respone electric signal of detection cell membrane sample under difficult environmental conditions becomes
Change, ion channel feature image and photoelectric respone electric signal image under the conditions of generation varying environment；Or
According to the ion channel pattern and photoelectric respone electric signal of the cell membrane sample, the conductive pinpoint positioning is driven
In the single ion channel of cell membrane sample, change cell membrane sample environment, the list of cell membrane sample is observed using scanning probe
The photoelectric respone change in electric of individual ion channel under difficult environmental conditions.
Second aspect, the embodiment of the present invention additionally provides a kind of cellular electrophysiologicalsensor detection dress for being excited based on light and scanning probe
Put, the device includes：
Probe actuation module is scanned, conductive pinpoint and the cell membrane on sample substrate for driving scanning probe
Sample is contacted；
Light excites performing module, for exciting the cell membrane sample using LASER Light Source so that the cell membrane sample
Obtain photoelectric characteristic response；
Applied electronic signal performing module, for by the conductive pinpoint to the cell membrane sample extrinsic motivated telecommunications
Number；
Pattern and photoelectric respone electric signal record performing module, for using cell membrane sample described in the scanning probe records
Ion channel pattern and photoelectric respone electric signal of the product in the case where light is excited.
Preferably, the light excite performing module specifically for：Regulate and control the optical maser wavelength and intensity of LASER Light Source, using not
The laser excitation cell membrane sample of co-wavelength and intensity；The pattern and electric signal record performing module specifically for：Using institute
State ion channel pattern and photoelectricity of the cell membrane sample under the laser excitation of different wave length and intensity described in scanning probe records
Respond electric signal.
Preferably, the device also includes：Pinpoint electric signal record performing module, for according to the cell membrane sample from
Subchannel pattern and photoelectric respone electric signal, drive the conductive pinpoint to be positioned at the ion channel fixed point of cell membrane sample, change
Become the wavelength and/or intensity of the LASER Light Source, pinpointed using the ion channel of scanning probe records cell membrane sample in difference
Wavelength and/or intensity laser excitation under local photoelectric respone change in electric.The third aspect, the embodiment of the present invention is also carried
Supplied it is a kind of based on light excite scanning probe cellular electrophysiologicalsensor detection system, including scanning probe, in addition to LASER Light Source and
Device described in second aspect, described device connects the scanning probe and LASER Light Source, the conduction of the scanning probe respectively
Needle point is located at the center for the laser that LASER Light Source is launched.
Preferably, the system also includes sample room, and the cell membrane sample, scanning probe and sample substrate are respectively positioned on described
Sample room, the sample room is connected with air and water pump, and vapor can be pumped into sample room by the air and water pump；Set in the sample room
There is temp-controlling element.
The present invention provide based on light excite scanning probe physiological detection cell membrane electrical characteristics and ion channel method,
Device and system, can obtain the channel current of cell membrane in real time directly while sample surfaces detect nanoscale pattern
And electrology characteristic, and sample structure and property are not destroyed.
Brief description of the drawings
Fig. 1 is the cellular electrophysiologicalsensor detection method flow that a kind of in the embodiment of the present invention one excites scanning probe based on light
Figure；
Fig. 2 is the cellular electrophysiologicalsensor detection method flow that a kind of in the embodiment of the present invention two excites scanning probe based on light
Figure；
Fig. 3 is the cellular electrophysiologicalsensor detection method flow that a kind of in the embodiment of the present invention three excites scanning probe based on light
Figure；
Fig. 4 is the cellular electrophysiologicalsensor detection device structure that a kind of in the embodiment of the present invention four excites scanning probe based on light
Schematic diagram；
Fig. 5 is the cellular electrophysiologicalsensor detection device structure that a kind of in the embodiment of the present invention five excites scanning probe based on light
Schematic diagram；
Fig. 6 is the cellular electrophysiologicalsensor detection device structure that a kind of in the embodiment of the present invention six excites scanning probe based on light
Schematic diagram；
Fig. 7 is the cellular electrophysiologicalsensor detection system structure that a kind of in the embodiment of the present invention seven excites scanning probe based on light
Schematic diagram；
Fig. 8 is the cellular electrophysiologicalsensor detection system knot that another in the embodiment of the present invention seven excites scanning probe based on light
Structure schematic diagram；
Fig. 9 is the cellular electrophysiologicalsensor detection system knot that another in the embodiment of the present invention seven excites scanning probe based on light
Structure schematic diagram.
Embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples.It is understood that this place is retouched
The specific embodiment stated is used only for explaining the present invention, rather than limitation of the invention.It also should be noted that, in order to just
Part related to the present invention rather than entire infrastructure are illustrate only in description, accompanying drawing.
Embodiment one
Fig. 1 is that a kind of of the offer of the embodiment of the present invention one excites the cellular electrophysiologicalsensor detection method for scanning probe based on light
Flow chart, the present embodiment is applicable to the situation of cell electrical characteristics and ion channel detection, for example, rice leaf cell electricity is special
Property and ion channel detection, crops/phytochrome is to cell ion channel and the regulatory mechanism of proton pump, opsin light
Quick ion channel, the neuronal cell that expressing has opsin can be activated specifically, and information is handed between research neuronal cell
Stream mechanism, draws neutral net etc., and this method can be by exciting the cellular electrophysiologicalsensor detection device of scanning probe to hold based on light
OK, the device can be realized by hardware and/or software, for example, can be the control unit for being integrated in scanning probe microscopic system
Or control program.As shown in figure 1, this method includes：
Step S110, the conductive pinpoint of driving scanning probe are contacted with the cell membrane sample on sample substrate.
Wherein, cell membrane sample is located on sample substrate, and scanning probe is located above cell membrane, driving scanning probe
A conductive pinpoint side contacts relative with sample substrate with cell membrane sample.
Step S120, the cell membrane sample excited using LASER Light Source so that it is special that the cell membrane sample obtains photoelectricity
Property response.
Wherein, the cell membrane sample is excited using LASER Light Source so that the cell membrane sample obtains photoelectric characteristic and rung
Should, the opened/closed degree that the ion channel of cell membrane sample is logical changes, and ion channel pattern and electric signal become
Change.
The cell membrane sample is excited to specifically include using LASER Light Source：Using laser excitation sample certain time.It is different
Cell sample obtains the influence of the time Stimulated Light firing time of photoelectric characteristic response, and the ion channel of different cell membrane samples is beaten
Laser firing time required for opening is different, and relevant with the photosensitive property of cell membrane sample, being excited to light for having is very sensitive, has
It is less sensitive.For example, bacteria rhodopsin cell membrane is only needed to several seconds.Those skilled in the art can select as needed
The time of laser excitation, the present invention is not especially limited to laser firing time.
Step S130, by the conductive pinpoint to the cell membrane sample extrinsic motivated electric signal；
Step S130, using it is described scanning probe records described in cell membrane sample light excite with extrinsic motivated electric signal
Ion channel pattern and photoelectric respone electric signal.
Wherein, using it is described scanning probe records described in cell membrane sample light excite with extrinsic motivated electric signal from
Subchannel pattern and electric signal are specifically included：The cell membrane sample is detected in the case where light is excited using the scanning probe scanning
Ion channel pattern, ion channel electric signal of the cell membrane sample in the case where light is excited is detected using the scanning probe.
In addition, the electric signal in order to be able to more accurately record ion channel, can also pass through the electric signal of ion channel
Preamplifier amplification is recorded again.
Preferably, using ion channel pattern and electricity of the cell membrane sample in the case where light is excited described in the scanning probe records
After signal, in addition to：According to the ion channel shape appearance figure of ion channel morphogenesis cell membrane sample, according to each ion channel
The electric signal distribution map of electric signal cellulation membrane sample.
It is pointed out that step S110 and step S120 order can be exchanged, the result of the present invention is had no effect on.
On the basis of above-mentioned technical proposal, the cell membrane sample is excited to specifically include using LASER Light Source：Regulation and control swash
The optical maser wavelength and intensity of radiant, using cell membrane sample described in the laser excitation of different wave length and varying strength；It is described to adopt
Specifically included with the ion channel pattern and electric signal of cell membrane sample described in the scanning probe records：Visited using the scanning
Pin records ion channel pattern and electric signal of the cell membrane sample under the laser excitation of different wave length and varying strength.
Because different photosensitive proteins have different photochromic sensitiveness, have to sensitive to blue light, have to green light sensitive, have
To red light sensitiveness.For example, the opsin in cone cell can be further divided into long wave according to the difference of its absorption spectrum ranges
Long opsin, medium wavelength opsin, short wavelength's opsin etc..L-type cone cell, M types cone cell and S type cone cells it is quick
It is respectively 565nm, 535nm and 440nm to feel peak value.When being detected to them, wavelength is respectively chosen as sensitive peak value, i.e.,
565nm, 535nm and 440nm.Laser intensity can adjust from 1mW to 100mW, suitably be adjusted according to the size of light excitation current
Light intensity.
Using the benefit of cell membrane sample described in the laser excitation of different wave length and varying strength be can detect it is various not
Same photosensitive protein, can detect different albumen corresponding light-dependent current and light sensitive characteristic.Those skilled in the art can root
According to the wavelength and intensity of selection laser is needed, the present invention is not especially limited to the wavelength and intensity of laser.
The cellular electrophysiologicalsensor detection method that scanning probe is excited based on light of the present embodiment, cell membrane ion can be detected simultaneously
Passage electrical characteristics and ion channel nanoscale three-dimensional appearance, and sample structure and property are not destroyed.
Embodiment two
Fig. 2 is refer to, Fig. 2 is that a kind of of the offer of the embodiment of the present invention two excites the cellular electrophysiologicalsensor for scanning probe based on light
The flow chart of detection method.Improvement is optimized based on above-described embodiment scheme in the present embodiment, and it is fixed in particular, provide
The concrete scheme of point detection.As shown in Fig. 2 this method includes：
Step S210, the conductive pinpoint of driving scanning probe are contacted with the cell membrane sample on sample substrate.
Step S220, the cell membrane sample excited using LASER Light Source so that it is special that the cell membrane sample obtains photoelectricity
Property response.
Step S230, by the conductive pinpoint to the cell membrane sample extrinsic motivated electric signal.
Step S240, using it is described scanning probe records described in cell membrane sample light excite with extrinsic motivated electric signal
Ion channel pattern and electric signal.
Step S250, ion channel pattern and photoelectric respone electric signal according to the cell membrane sample, driving described in lead
Acusector point is positioned at the ion channel fixed point of cell membrane sample, changes the wavelength and/or intensity of the LASER Light Source, using scanning
The local photoelectricity that the ion channel of probe records cell membrane sample is pinpointed under the laser excitation of different wavelength and/or intensity rings
Answer change in electric.
Wherein, drive the conductive pinpoint to be positioned at the ion channel fixed point of cell membrane sample, change the LASER Light Source
Wavelength and/or intensity, using scanning probe detection cell membrane sample ion channel pinpoint in different wavelength and/or intensity
Laser excitation under local electric signal so that according to the local telecommunications under the laser excitation of different wavelength and/or intensity
Number, obtain the ion channel electrical characteristics curve of cell membrane sample.
The present embodiment uses the detection mode that contact type scanning and fixed point detection are combined.Contact type scanning, is utilized
Conductive pinpoint scanning probe cell membrane sample ion channel pattern and electric signal；Fixed point detection, according to ion channel pattern and electricity
Signal, is positioned at ion channel in the sample of scanning using needle point and pinpoints, the local electric current of ion channel is measured and
Imaging.So as to directly sample surfaces detect nanoscale pattern while, in real time obtain cell membrane channel current and
Electrology characteristic, and sample structure and property are not destroyed.
Embodiment three
Fig. 3 is refer to, Fig. 3 is a kind of cellular electrophysiologicalsensor detection side based on scanning probe that the embodiment of the present invention three is provided
The flow chart of method.Improvement is optimized based on above-described embodiment scheme in the present embodiment, in particular, provides change sample
Environment is scanned the concrete scheme of detection or fixed point detection.As shown in figure 3, this method includes：
Step S310, the conductive pinpoint of driving scanning probe are contacted with the cell membrane sample on sample substrate.
Step S320, the cell membrane sample excited using LASER Light Source so that it is special that the cell membrane sample obtains photoelectricity
Property response.
Step S330, by the conductive pinpoint to the cell membrane sample extrinsic motivated electric signal.
Step S340, using ion channel pattern of the cell membrane sample in the case where light is excited described in the scanning probe records and
Photoelectric respone electric signal.
Step S350, change cell membrane sample environment, the photoelectric respone of detection cell membrane sample under difficult environmental conditions
Ion channel feature image and photoelectric respone electric signal image under the conditions of change in electric, generation varying environment；Or
According to the ion channel pattern and photoelectric respone electric signal of the cell membrane sample, the conductive pinpoint positioning is driven
In the single ion channel of cell membrane sample, change cell membrane sample environment, the list of cell membrane sample is observed using scanning probe
The photoelectric respone change in electric of individual ion channel under difficult environmental conditions.
Wherein, changing cell membrane sample environment includes medicine, humidity or temperature etc., on the one hand, scanned by conductive pinpoint
Ion under the conditions of the photoelectric respone change in electric of detection cell membrane sample under difficult environmental conditions, generation varying environment leads to
Road feature image and photoelectric respone electric signal image, what is now obtained is cell membrane sample entirety or the ion channel in some region
Feature image and photoelectric respone electric signal image；On the other hand, the conductive pinpoint is driven to be positioned at the single of cell membrane sample
Ion channel, to observe the photoelectric respone change in electric of single ion channel under difficult environmental conditions, what is now obtained is
The single ion channel photoelectric respone change in electric of cell membrane sample.
The electric signal opened by each ion channel, can directly obtain electric signal amplitude distribution that each ion channel opens,
The function parameter such as probability and open life-span distribution is opened, on this basis acceptable and analytic function parameter and film potential, ion
Relation between concentration etc..
The present embodiment uses the detection mode that contact type scanning and fixed point detection are combined, can be directly in sample table
While face detects nanoscale pattern, the channel current and electrology characteristic of cell membrane are obtained in real time, and do not destroy sample knot
Structure and property.
The following is provided in the specific embodiment of the invention it is a kind of based on light excite scanning probe cellular electrophysiologicalsensor visit
The embodiment of device is surveyed, embodiment of the embodiment based on above-mentioned method of device realizes that not most description in a device please be joined
Examine the embodiment of preceding method.
Example IV
Fig. 4 is refer to, Fig. 4 is that a kind of of fourth embodiment of the invention offer excites the cell electricity of scanning probe raw based on light
Manage the structural representation of detection device.As shown in figure 4, the device 1 includes：
Probe actuation module 101 is scanned, for driving the conductive pinpoint of scanning probe thin on sample substrate with being located at
After birth sample is contacted；
Light excites performing module 102, for exciting the cell membrane sample using LASER Light Source so that so that the cell
Membrane sample obtains photoelectric characteristic response；
Applied electronic signal performing module 103, for electric to the cell membrane sample extrinsic motivated by the conductive pinpoint
Signal；
Pattern and photoelectric respone electric signal record performing module 104, for using cell described in the scanning probe records
Ion channel pattern and electric signal of the membrane sample in the case where light is excited.
Further, the light excite performing module 102 specifically for：Regulate and control the optical maser wavelength and intensity of LASER Light Source,
Using different wave length and the laser excitation cell membrane sample of intensity；The pattern and photoelectric respone electric signal record performing module
104 specifically for：Using cell membrane sample described in the scanning probe records under the laser excitation of different wave length and intensity
Ion channel pattern and electric signal.
Connected each other between each module of the present embodiment and realize that contact type scanning is detected, directly can received in sample surfaces detection
While metrical scale pattern, the channel current and electrology characteristic of cell membrane are obtained in real time, and do not destroy sample structure and property.
Embodiment five
Improvement is optimized based on the scheme of example IV in embodiment five, in particular, provides fixed point detection
Scheme, the difference is that only fixed point pattern many and electric signal record performing module 104 with example IV.As shown in figure 5, should
Device 1 also includes：
Electric signal record performing module 105 is pinpointed, for the ion channel pattern and photoelectricity according to the cell membrane sample
Electric signal is responded, drives the conductive pinpoint to be positioned at the ion channel fixed point of cell membrane sample, changes the LASER Light Source
Wavelength and/or intensity, are pinpointed in different wavelength and/or intensity using the ion channel of scanning probe records cell membrane sample
Local photoelectric respone change in electric under laser excitation.
Embodiment six
Improvement is optimized based on example IV or five scheme in embodiment six, in particular, provides change sample
Product environment is scanned the scheme of detection or fixed point detection, the difference is that only more the signal detection under single environmental Kuznets Curves
Module 106.As shown in fig. 6, the device 1 also includes：
Signal detection module 106 under environmental Kuznets Curves, is used for：
Change cell membrane sample environment, the photoelectric respone electric signal of detection cell membrane sample under difficult environmental conditions becomes
Change, ion channel feature image and photoelectric respone electric signal image under the conditions of generation varying environment；Or
According to the ion channel pattern and photoelectric respone electric signal of the cell membrane sample, the conductive pinpoint positioning is driven
In the single ion channel of cell membrane sample, change cell membrane sample environment, the list of cell membrane sample is observed using scanning probe
The photoelectric respone change in electric of individual ion channel under difficult environmental conditions.
Connect the detection for realizing that contact type scanning and fixed point detection are combined between each module of embodiment five and six each other
Pattern, can obtain the channel current and electricity of cell membrane in real time directly while sample surfaces detect nanoscale pattern
Characteristic, and sample structure and property are not destroyed.
Embodiment seven
Refer to Fig. 7-9, Fig. 7-9 is that a kind of light that seventh embodiment of the invention is provided excites the cell based on scanning probe
The structural representation of electro physiology detection system.As shown in figures 7 and 9, the system includes：Above-described embodiment four-six is any described
Device 1, scanning probe 2 and LASER Light Source 3, described device 1 connects the scanning probe 2 and LASER Light Source 3 respectively, described
The conductive pinpoint for scanning probe 2 is located at the center for the laser that LASER Light Source 3 is launched.
Specifically, the control end connection described device 1 of the scanning probe 1, the scanning probe 1 is in described device 1
Driving is lower to move, and is contacted with cell membrane sample, ion channel pattern of the scanning record detection cell membrane sample in the case where light is excited with
Electric signal.
The needle point for scanning probe is conducting probe, and body construction is silica or silicon nitride material, and needle point coating is conductive
Layer, conductive layer is golden film, chromium, titanium, platinoiridita laminated film etc..
LASER Light Source 3 includes optical maser wavelength and the adjustable laser generator of luminous intensity and right-angled triangular prism.Laser from
Laser generator horizontal exit, 90 degree of directive cell membrane samples 6 straight up are turned to by triangular prism reflection.The laser light
The control end connection described device 1 of the laser generator in source 3.The photoelectricity that the LASER Light Source 3 is used for activated cell membrane sample is special
Property response.The LASER Light Source 3 is opened or closed under the control of described device 1, adjustment optical maser wavelength and intensity.In order to
Make laser directive cell membrane sample 6 straight up, the sample substrate 7 for placing the cell membrane sample 6 is printing opacity graphite base.
On the basis of the technical scheme of above-described embodiment, as shown in Figure 8 and Figure 9, the system also include air and water pump 4,
Temp-controlling element 5 and sample room 8, the scanning probe 2, cell membrane sample 6 and sample substrate 7 are respectively positioned on the sample room 8, institute
State sample room 8 with air and water pump 4 to be connected, vapor can be pumped into the sample room 8 by the air and water pump 4；Set in the sample room 8
There is temp-controlling element 5.The control end of air and water pump 4 and temp-controlling element 5 connects described device 1 respectively, changes under the driving of described device 1
Become cell sample loop border.
Specifically, the sample room 8 is the sealing device with moisture outlet and moisture ingress, moisture ingress and air and water pump 4
It is connected, vapor can be pumped into sample room 8 and adjust humidity by air and water pump 4, keeps the bioactivity of cell membrane to be measured.Temp-controlling element
5 be semiconductor refrigerating element, and control sample temperature is maintained in less than 10 degree of low temperature environment, keeps the biology of biomembrane to be measured
Activity.
The said goods can perform the method that any embodiment of the present invention is provided, and possess the corresponding functional module of execution method
And beneficial effect.
Note, above are only presently preferred embodiments of the present invention and institute's application technology principle.It will be appreciated by those skilled in the art that
The invention is not restricted to specific embodiment described here, can carry out for a person skilled in the art it is various it is obvious change,
Readjust and substitute without departing from protection scope of the present invention.Therefore, although the present invention is carried out by above example
It is described in further detail, but the present invention is not limited only to above example, without departing from the inventive concept, also
Other more equivalent embodiments can be included, and the scope of the present invention is determined by scope of the appended claims.
Claims (10)
1. a kind of excite the cellular electrophysiologicalsensor detection method for scanning probe based on light, it is characterised in that this method includes：
The conductive pinpoint of driving scanning probe is contacted with the cell membrane sample on sample substrate, is excited using LASER Light Source
The cell membrane sample so that the cell membrane sample obtains photoelectric characteristic response；
By the conductive pinpoint to the cell membrane sample extrinsic motivated electric signal；
Excited and the ion channel shape under extrinsic motivated electric signal in light using cell membrane sample described in the scanning probe records
Looks and photoelectric respone electric signal.
2. according to the method described in claim 1, it is characterised in that existed using cell membrane sample described in the scanning probe records
Light is excited to be specifically included with the ion channel pattern and electric signal under extrinsic motivated electric signal：Visited using the scanning probe scanning
Ion channel pattern of the cell membrane sample under photo-excitation conditions is surveyed, the cell membrane sample is detected using the scanning probe
The photoelectric respone electric signal of ion channel of the product under photo-excitation conditions；
Excited and the ion channel shape under extrinsic motivated electric signal in light using cell membrane sample described in the scanning probe records
After looks and photoelectric respone electric signal, in addition to：According to the ion channel shape appearance figure of ion channel morphogenesis cell membrane sample,
According to the photoelectric respone electric signal distribution map of the photoelectric respone electric signal cellulation membrane sample of each ion channel.
3. according to the method described in claim 1, it is characterised in that excite the cell membrane sample specifically to wrap using LASER Light Source
Include：Regulate and control the optical maser wavelength and intensity of LASER Light Source, using cell membrane sample described in the laser excitation of different wave length and varying strength
Product；Excited and the ion channel pattern under extrinsic motivated electric signal in light using cell membrane sample described in the scanning probe records
Specifically included with photoelectric respone electric signal：Using it is described scanning probe records described in cell membrane sample different wave length with it is difference strong
Ion channel pattern and photoelectric respone electric signal under the laser excitation of degree.
4. according to the method described in claim 1, it is characterised in that existed using cell membrane sample described in the scanning probe records
Light excite with after the ion channel pattern and photoelectric respone electric signal under extrinsic motivated electric signal, in addition to：
According to the ion channel pattern and photoelectric respone electric signal of the cell membrane sample, the conductive pinpoint is driven to be positioned at carefully
The ion channel fixed point of after birth sample, changes the wavelength and/or intensity of the LASER Light Source, using scanning probe records cell membrane
The ion channel of sample pinpoints the local photoelectric respone change in electric under the laser excitation of different wavelength and/or intensity.
5. according to the method described in claim 1, it is characterised in that existed using cell membrane sample described in the scanning probe records
Light excite with after the ion channel pattern and photoelectric respone electric signal under extrinsic motivated electric signal, in addition to：
Change cell membrane sample environment, the photoelectric respone change in electric of detection cell membrane sample under difficult environmental conditions is raw
Into the ion channel feature image under the conditions of varying environment and photoelectric respone electric signal image；Or
According to the ion channel pattern and photoelectric respone electric signal of the cell membrane sample, the conductive pinpoint is driven to be positioned at carefully
The single ion channel of after birth sample, change cell membrane sample environment, using scanning probe observation cell membrane sample it is single from
The photoelectric respone change in electric of subchannel under difficult environmental conditions.
6. a kind of excite the cellular electrophysiologicalsensor detection device for scanning probe based on light, it is characterised in that the device includes：
Probe actuation module is scanned, conductive pinpoint and the cell membrane sample on sample substrate for driving scanning probe
Contact；
Light excites performing module, for exciting the cell membrane sample using LASER Light Source so that the cell membrane sample is obtained
Photoelectric characteristic is responded；
Applied electronic signal performing module, for by the conductive pinpoint to the cell membrane sample extrinsic motivated electric signal；
Pattern and photoelectric respone electric signal record performing module, for being existed using cell membrane sample described in the scanning probe records
Light excite under ion channel pattern and photoelectric respone electric signal.
7. device according to claim 6, it is characterised in that the light excite performing module specifically for：Regulate and control laser
The optical maser wavelength and intensity of light source, using different wave length and the laser excitation cell membrane sample of intensity；The pattern and electric signal
Record performing module specifically for：Using cell membrane sample described in the scanning probe records in different wave length and the laser of intensity
Ion channel pattern and photoelectric respone electric signal under exciting.
8. device according to claim 6, it is characterised in that the device also includes：Electric signal record performing module is pinpointed,
For the ion channel pattern and photoelectric respone electric signal according to the cell membrane sample, the conductive pinpoint is driven to be positioned at carefully
The ion channel fixed point of after birth sample, changes the wavelength and/or intensity of the LASER Light Source, using scanning probe records cell membrane
The ion channel of sample pinpoints the local photoelectric respone change in electric under the laser excitation of different wavelength and/or intensity.
9. the cellular electrophysiologicalsensor detection system of scanning probe is excited based on light a kind of, including scans probe, it is characterised in that is also wrapped
LASER Light Source and the device described in claim any one of 6-8 are included, described device connects the scanning probe and laser light respectively
Source, the conductive pinpoint of the scanning probe is located at the center for the laser that LASER Light Source is launched.
10. system according to claim 9, it is characterised in that the system also includes sample room, the cell membrane sample,
Scanning probe is respectively positioned on the sample room with sample substrate, and the sample room is connected with air and water pump, and the air and water pump can steam water
Air pump enters sample room；Temp-controlling element is provided with the sample room.
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Application publication date: 20171107 |