CN113721043B - SICM scanning system and method based on array line laser - Google Patents
SICM scanning system and method based on array line laser Download PDFInfo
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- CN113721043B CN113721043B CN202111016831.4A CN202111016831A CN113721043B CN 113721043 B CN113721043 B CN 113721043B CN 202111016831 A CN202111016831 A CN 202111016831A CN 113721043 B CN113721043 B CN 113721043B
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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
- G01Q60/44—SICM [Scanning Ion-Conductance Microscopy] or apparatus therefor, e.g. SICM probes
Abstract
The invention discloses a SICM scanning system and method based on array line laser, wherein an array line laser emission source and a CCD photosensitive camera are respectively arranged at two sides of an electrochemical cell, a laser beam generated by the array line laser emission source penetrates through the electrochemical cell and is applied to the CCD photosensitive camera, the height of a scanning sample to be detected in the electrochemical cell can be rapidly adjusted, the height of a glass probe tube is determined by utilizing the laser beam, the highest jumping position of a scanning ion conductance microscope in a jumping scanning mode can be rapidly determined, the sample scanning efficiency is greatly improved, and the scanning time cost is reduced; after the imaging of the array line laser is processed and analyzed by the image processing unit, the node feedback is connected with the hardware system controller to obtain the highest point position of the sample, the closed-loop feedback system optimizes the traditional sample scanning process to a great extent, the time cost is saved in sample scanning, and the closed-loop feedback system has great effect and significance in capturing the cell pharmacological reaction morphology change in the life science field.
Description
Technical Field
The invention belongs to the field of scanning ion conductance microscopes, and particularly relates to a SICM scanning system and method based on array line laser.
Background
The Scanning Ion Conductance Microscope (SICM) as a micro-imaging tool emerging in recent years has a great application prospect in the field of micro-imaging, and is particularly suitable for carrying out shape imaging on a sample material in a submerged environment. The probe microscope has no contact influence on a sample in the scanning process, can detect the original state of the sample in real time, quickly and at high resolution, has great advantage particularly in detecting the sample with soft surface, and is widely applied to the fields of micro-nano biological cells, new materials, medicine, pharmacy and the like. Compared with a scanning atomic force microscope and a scanning electron microscope, the scanning atomic force microscope and the scanning electron microscope are slightly inferior in precision, but the scanning atomic force microscope and the scanning electron microscope can still achieve micron-sized scanning effect, have functions which are not provided by the microscopes, such as the effects of physicochemical analysis, material delivery and extraction, and have great application prospects in scientific research.
With the continuous development of the technology, the scanning ion conductance microscope has undergone multiple improvements, which are further improved from the first generation in principle and structure, but the improvement of the performance of the instrument is always the key point for improving the scanning speed. Related researchers at home and abroad only start with the scanning principle to improve the scanning speed, and include methods such as a prediction scanning method, a compressed sensing scanning method, a two-step downward probing scanning method and the like, which improve the scanning speed of an instrument to a certain extent, but are not widely applied to the sample scanning process in the actual process, so further exploration and improvement are needed in the aspect; the research work on the performance of SICM imaging technology instruments is relatively very short nowadays, and particularly in China, the research on instrument development and application is very little, and the SICM imaging technology instruments are all made of the phoenix feather bone.
The conventional scanning ion conductance microscope generally adopts a traditional jump scanning mode, the first problem faced by the conventional scanning ion conductance microscope is how to determine the jump height of a probe, the jump height is generally set manually by experience, the jump height directly determines the scanning time of a sample, if the jump height is too large, the scanning speed is slow, and if the jump height is too small, the probe tip is contacted with the surface of the sample, so that the scanning is interrupted. Therefore, a quick and stable imaging method for determining the jumping peak by self-adapting to the target area of the sample to be detected is lacked at present.
Disclosure of Invention
The invention aims to provide a SICM scanning system and a SICM scanning method based on array line laser, which are used for overcoming the defects of the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a SICM scanning system based on array line laser comprises an ion current collector, a glass probe tube, a micro-displacement control console, an array line laser emission source, a CCD photosensitive camera and a core control unit; the glass probe tube is fixed on the micro-displacement control console, the glass probe tube can extend into an electrochemical cell provided with a sample to be scanned, one end of the ion current collector is connected with the reference electrode through a power supply, the other end of the ion current collector is connected with the detection electrode, and the detection electrode is arranged in the glass probe tube; the array line laser emission source and the CCD photosensitive camera are respectively arranged on two sides of the electrochemical cell, a laser beam generated by the array line laser emission source penetrates through the electrochemical cell and strikes the CCD photosensitive camera, the CCD photosensitive camera is connected with an image processing unit, and the image processing unit, the micro-displacement control console and the ionic current collector are all connected to the core control unit.
Furthermore, the micro-displacement control platform comprises a Z-direction micro-motor, Z-direction piezoelectric ceramics and XY-direction piezoelectric ceramics, wherein the XY-direction piezoelectric ceramics are arranged on the Z-direction micro-motor, the Z-direction piezoelectric ceramics are fixed on one side of the XY-direction piezoelectric ceramics, and the glass probe tube is fixed on one side of the Z-direction piezoelectric ceramics.
Further, array line laser emission source fixed mounting is on finely tuning the platform, finely tune the platform and include vertical direction fine setting screw rod and horizontal fine setting structure, horizontal fine setting structure includes array line laser fine setting platform base, array line laser fine setting platform base is provided with can follow its lateral sliding's horizontal base, be provided with vertical slide on the horizontal base, array line laser emission source is fixed in on the vertical slide, be equipped with vertical spout on the horizontal base, vertical slide sets up in vertical spout, vertical direction fine setting screw rod passes through threaded connection with vertical slide.
Furthermore, the transverse base and the array line laser fine tuning platform base slide in a matched mode through a dovetail groove, and a locking knob used for locking the transverse base is arranged on one side of the array line laser fine tuning platform base.
Furthermore, the glass probe tube is a conical hollow tube, and the diameter of the opening at the tip end of the glass probe tube is less than or equal to 100nm.
Furthermore, the core control unit comprises an FPGA control chip, a high-speed AD conversion chip for converting the ion current in the loop in real time, a driving module for driving the micro motor and the piezoelectric ceramics and a power module, and the FPGA control chip is used for outputting a control signal to the driving module.
Furthermore, an Ag/AgCl electrode is adopted as a reference electrode.
Furthermore, the Z-direction movement range of the Z-direction piezoelectric ceramic is less than or equal to 30 microns, the XY-direction movement range of the XY-direction piezoelectric ceramic is less than or equal to 100 microns, and the precision is +/-1 nm.
Furthermore, the movement range of the Z-direction micro motor is less than or equal to 15mm, and the precision is +/-1 mu m.
A SICM scanning method comprises the following steps:
s1, calibrating the positions of an array line laser emission source and a CCD photosensitive camera, so that the CCD photosensitive camera can receive complete laser beam imaging;
s2, adjusting the height of the electrochemical cell until the lower end of the laser beam image received by the CCD photosensitive camera is incomplete, and stopping height adjustment of the electrochemical cell;
and S3, adjusting the height of the glass probe tube, acquiring laser beam imaging in real time until the upper end of the laser beam imaging is incomplete, acquiring the highest point position information of the sample, setting the jump height information of the scanning ion conductance microscope in a jump scanning mode, and scanning the surface of the sample based on the highest point position information.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention relates to a SICM scanning system based on array line laser, which comprises an ionic current collector, a glass probe tube, a micro-displacement control console, an array line laser emission source, a CCD photosensitive camera and a core control unit, wherein the array line laser emission source and the CCD photosensitive camera are respectively arranged at two sides of an electrochemical cell; after the array line laser is started, the laser enters the solution through the transparent wall surface of the electrochemical cell, penetrates out of the other side wall surface to reach the CCD photosensitive element, the imaging of the array line laser is processed and analyzed through the image processing unit, the junction feedback is connected with the hardware system controller, and the hardware system controller controls the corresponding executing mechanism according to the control mechanism to obtain the highest point position of the sample. The closed-loop feedback system greatly optimizes the traditional sample scanning process, particularly, on the aspect of determining the jump height position in the jump scanning mode, the time cost is saved on sample scanning, and the closed-loop feedback system has great effect and significance on capturing the change of the cell pharmacological reaction morphology in the field of life science.
The invention discloses a SICM scanning method, which starts from the actual scanning principle, uses an array line laser method, assists and optimizes the scanning process from the physical and physical perspective by improving the peripheral equipment of a scanning ion conductance microscope structure, and reduces the jump height in a jump scanning mode to the maximum extent by searching the highest point position of the sample morphology, thereby realizing the rapid scanning effect of the scanning ion conductance microscope.
Drawings
FIG. 1 is a schematic diagram of a scanning system according to an embodiment of the present invention.
FIG. 2 is a diagram of the case where the array laser does not contact the sample in the embodiment of the present invention.
FIG. 3 is a schematic diagram of determining a location of a highest point of a sample in an embodiment of the invention.
Fig. 4 is a schematic perspective view of the installation of the scanning system in the embodiment of the invention.
Fig. 5 is a flowchart of the operation of the scanning method in the embodiment of the present invention.
In the figure, 1-reference electrode; 2-a power supply; 3-a glass probe tube; 4-an ionic current collector; 5-Z direction piezoelectric ceramics; 6-XY piezoelectric ceramics; a 7-Z upward micromotor; 8-core control unit; 9-an image processing unit; a 10-Z down micromotor; 11-a CCD light-sensitive camera; 12-line laser beam; 13-a sample to be scanned; 14-an electrochemical cell; 15-array line laser emission source; 16-laser beam without contacting the sample; 17-a laser beam when contacting the sample and probe tip; 18-vertical direction fine adjustment knob; 19-a base; 20-horizontal direction fine adjustment structure; 21-array line laser fine tuning platform base; 22-a transverse base; 23-vertical slide.
Detailed Description
The invention is described in further detail below with reference to the accompanying drawings:
a SICM scanning system based on array line laser comprises an ion current collector 4, a glass probe tube 3, a micro-displacement control console, an array line laser emission source 15, a CCD photosensitive camera 11 and a core control unit 8; the glass probe tube 3 is fixed on the micro-displacement control console, the glass probe tube 3 can extend into the electrochemical cell 14, one end of the ionic current collector 4 is connected with the reference electrode through the power supply 2, the other end is connected with the detection electrode, and the detection electrode is arranged in the glass probe tube 3; the array line laser emission source 15 and the CCD light sensing camera 11 are respectively arranged on two sides of the electrochemical cell 14, a sample 13 to be scanned is placed in the electrochemical cell 14, a laser beam generated by the array line laser emission source 15 penetrates from the electrochemical cell 14 to the CCD light sensing camera 11, the axis of the glass probe tube 3 is perpendicular to the laser beam, the CCD light sensing camera 11 is connected with the image processing unit 9, and the image processing unit 9, the micro-displacement control console and the ionic current collector 4 are all connected to the core control unit 8.
The micro-displacement control platform comprises a Z-direction micro-motor 7, Z-direction piezoelectric ceramics 5 and XY-direction piezoelectric ceramics 6, the XY-direction piezoelectric ceramics 6 are arranged on the Z-direction micro-motor 7, the Z-direction piezoelectric ceramics 5 are fixed on one side of the XY-direction piezoelectric ceramics 6, and the glass probe tube 3 is fixed on one side of the Z-direction piezoelectric ceramics 5; the core control unit 8 controls the movement of the Z-direction piezoelectric ceramic 5 and the XY-direction piezoelectric ceramic 6 through the piezoelectric controller, and controls the up-and-down movement of the Z-direction micro-motor 7. The micro-displacement console is used for fine-tuning the glass probe tube 3. An electrochemical cell 14 is secured to Z-down micro-machine 10 for regulating vertical movement of electrochemical cell 14.
The array line laser emission source 1 is fixedly arranged on the fine adjustment platform, and the fine adjustment platform is used for adjusting the posture of the array line laser emission source; the fine setting platform includes vertical direction fine setting screw 18 and horizontal fine setting structure 20, horizontal fine setting structure 20 is including array line laser fine setting platform base 21, array line laser fine setting platform base 21 is provided with can follow its lateral sliding's horizontal base 22, be provided with vertical slide 23 on the horizontal base 22, array line laser emission source 15 is fixed in on vertical slide 23, be equipped with vertical spout on the horizontal base 2, vertical slide 23 sets up in vertical spout, vertical direction fine setting screw 18 passes through threaded connection with vertical slide 23, through rotatory vertical direction fine setting screw 18, realize vertical fine setting of vertical slide 23, drive the ascending removal of array line laser emission source 15 vertical side simultaneously.
The transverse base 22 and the array line laser fine tuning platform base 21 slide in a matched mode through a dovetail groove, and a locking knob used for locking the transverse base 22 is arranged on one side of the array line laser fine tuning platform base 21. The horizontal movement depends on a gear rack as a transmission mechanism, the dovetail grooves are used as a guide mechanism, the screw rod rack is used as a transmission mechanism in the vertical movement direction, the dovetail grooves are also used as a guide mechanism, and the dovetail grooves in the vertical direction are composed of two groups and are respectively distributed on two sides of the array line laser.
As shown in fig. 4, which is a schematic view of a scanning system mounting three-dimensional structure, a vertical fine adjustment knob 18 is used for adjusting the Z-direction position of array line laser, and a horizontal fine adjustment structure 20 is used for adjusting the X-direction position of an array line laser source, so as to ensure that the array laser emission source is at a proper position; in addition, the CCD light-sensitive camera 11 is used for receiving array line laser beams and transmitting imaging information to the image processing unit 9, wherein the CCD light-sensitive camera is fixed on the base, and the light-sensitive plane of the CCD light-sensitive camera is always vertical to the transmission direction of the array line laser beams so as to ensure the effectiveness and the accuracy of measurement;
the image processing unit 9 is used for processing the image of the array line laser beam on the CCD photosensitive camera 11 and sending the processed image to the core control unit 8;
as shown in fig. 1, the system core control unit 8 is used for controlling the piezoelectric ceramic controller and the micro-motor controller, and detecting the magnitude of the ion current.
The reference electrode 1 adopted by the ion current loop adopts an Ag/AgCl electrode;
the glass probe tube 3 is a conical hollow tube, as shown in fig. 1, the diameter of the opening at the tip is less than or equal to 100nm, and the solution filled in the glass probe tube is consistent with the solution in the electrochemical cell;
the core control unit 8 comprises an FPGA control chip, a high-speed AD conversion chip for converting the ion current in the loop in real time, a driving module for driving the micromotor and the piezoelectric ceramics and a power module; the Z-direction piezoelectric ceramic 5 and the XY-direction piezoelectric ceramic 6 are used for providing small-range high-precision positioning, wherein the Z-direction movement range is less than or equal to 30 micrometers, the XY-direction movement range is less than or equal to 100 micrometers, and the precision is +/-1 nm, and the Z-direction micro-motor 7 and the Z-direction micro-motor 10 can provide large-range movement, wherein the movement range is less than or equal to 15mm, and the precision is +/-1 micrometer;
the following is a specific working process of the invention:
as shown in fig. 1, in order to ensure that the array line laser emission source 15 and the CCD photo camera 11 are correctly matched, the array line laser emission source 15 is finely adjusted; the fine adjustment platform has two degrees of freedom, and respectively controls the vertical position and the horizontal X-direction position of the array line laser emission source, the specific structure is as shown in FIG. 4, and the corresponding fine adjustment of the position of the array line laser emission source can be realized by respectively operating two knobs;
firstly, starting from a first state, namely that an array line laser beam is not contacted with the highest point of a sample, imaging of the laser beam on a CCD photosensitive camera 11 is complete at the moment, as shown in the condition of FIG. 2, at the moment, after the acquired image is processed by an image processing unit 9, the processing result is fed back to a system core controller 8, according to the working process, the core controller 8 controls a Z-down micro-motor 10 through controlling a micro-motor controller, so that an electrochemical cell 14 moves upwards along the vertical direction, namely, a sample 13 to be scanned also moves upwards at the same time until the highest point of the sample 13 to be scanned is contacted with a certain position of the array line laser, at the moment, as the sample 13 to be detected is a non-transparent substance, the array line laser is partially shielded on the Z-direction width, imaging of the array line laser on the CCD photosensitive camera 11 can be changed, the information can be captured through the image processing unit 9, and then the Z-down micro-motor 10 stops moving by utilizing a feedback mechanism, at the moment, the system reaches a second state, as shown in the position of the array line laser beam shown in FIG. 1;
then, preparing to enter a third state, wherein the strokes of the piezoelectric ceramics 5 and 6 are very small and are only suitable for high-precision feeding in a small range, so that the piezoelectric ceramics are required to act together with the micro-motor 7 in the Z direction movement process; in order to determine the jump height position of the scanning ion conductance microscope probe in the jump scanning mode, the system core controller 8 controls the microcomputer controller to control the Z-direction micro-motor 7 to move, so that the probe moves downwards until the tip of the glass probe tube 3 contacts the plane formed by the array line laser beams, at this time, the laser beams generate corresponding refraction at the tip of the glass probe tube 3, so that the image on the CCD photo-sensing camera 11 changes again, as in the previous similar way, the change information can be captured by the image processing unit 9, and then the Z-direction micro-motor 7 stops moving by using the feedback mechanism again, at this time, the system reaches a third state, as shown in fig. 3, the highest point position in the Z direction is recorded here and is used as the jump height in the jump scanning mode;
finally, the jump highest point position is provided, and the appearance scanning of the sample is carried out according to the jump scanning mode of the scanning ion conductance microscope, because the optimal jump height is realized by the method and is not set according to experience, the Z downward probing distance is greatly reduced, and the phenomenon of 'firing pin' caused by insufficient jump height setting in the past is also avoided;
the invention realizes the purpose of fast scanning the sample by the scanning ion conductance microscope based on the array line laser, thereby greatly saving the scanning time cost and believing that the scanning ion conductance microscope has wide prospect in the subsequent scientific research.
Claims (8)
1. A SICM scanning system based on array line laser is characterized by comprising an ion current collector (4), a glass probe tube (3), a micro-displacement control console, an array line laser emission source (15), a CCD photosensitive camera (11) and a core control unit (8); the glass probe tube (3) is fixed on the micro-displacement control console, the glass probe tube (3) can extend into an electrochemical cell (14) provided with a sample (13) to be scanned, one end of the ionic current collector (4) is connected with a reference electrode through a power supply (2), the other end of the ionic current collector is connected with a detection electrode, and the detection electrode is arranged in the glass probe tube (3); the array line laser emission source (15) and the CCD photosensitive camera (11) are respectively arranged on two sides of the electrochemical cell (14), a laser beam generated by the array line laser emission source (15) penetrates through the electrochemical cell (14) and is applied to the CCD photosensitive camera (11), the CCD photosensitive camera (11) is connected with an image processing unit (9), and the image processing unit (9), the micro-displacement control console and the ion current collector (4) are all connected with the core control unit (8); the array line laser emission source (15) is fixedly installed on the fine adjustment platform, the fine adjustment platform comprises a vertical fine adjustment screw (18) and a horizontal fine adjustment structure (20), the horizontal fine adjustment structure (20) comprises an array line laser fine adjustment platform base (21), the array line laser fine adjustment platform base (21) is provided with a transverse base (22) capable of sliding transversely along the array line laser fine adjustment platform base, the transverse base (22) is provided with a vertical sliding seat (23), the array line laser emission source (15) is fixed on the vertical sliding seat (23), the transverse base (22) is provided with a vertical sliding groove, the vertical sliding seat (23) is arranged in the vertical sliding groove, and the vertical fine adjustment screw (18) is connected with the vertical sliding seat (23) through threads; the Z-direction movement range of the Z-direction piezoelectric ceramic (5) is less than or equal to 30 mu m, the XY-direction movement range of the XY-direction piezoelectric ceramic (6) is less than or equal to 100 mu m, and the precision is +/-1 nm.
2. The SICM scanning system based on array line laser according to claim 1, wherein said micro displacement control platform comprises a Z direction micro motor (7), a Z direction piezoelectric ceramic (5) and an XY direction piezoelectric ceramic (6), the XY direction piezoelectric ceramic (6) is arranged on the Z direction micro motor (7), the Z direction piezoelectric ceramic (5) is fixed on one side of the XY direction piezoelectric ceramic (6), the glass probe tube (3) is fixed on one side of the Z direction piezoelectric ceramic (5).
3. The SICM scanning system based on array line laser as claimed in claim 1, wherein the transverse base (22) and the array line laser fine tuning platform base (21) slide in a dovetail groove fit manner, and a locking knob for locking the transverse base (22) is arranged on one side of the array line laser fine tuning platform base (21).
4. The SICM scanning system based on array line laser according to claim 1, characterized in that the glass probe tube (3) is a tapered hollow tube with tip opening diameter less than or equal to 100nm.
5. The SICM scanning system based on array line laser as claimed in claim 1, wherein the core control unit (8) includes FPGA control chip, high speed AD conversion chip for real-time conversion of ion current in the loop, driving module for driving micro-motor and piezoelectric ceramic and power module, the FPGA control chip is used to output control signal to the driving module.
6. The SICM scanning system based on array line laser as claimed in claim 1, wherein the reference electrode (1) is Ag/AgCl electrode.
7. The SICM scanning system based on array line laser as claimed in claim 1, wherein the micro-motor (7) in Z direction has a motion range of 15mm or less and a precision of ± 1 μm.
8. A SICM scanning method based on the SICM scanning system based on array line laser according to claim 1, comprising the steps of:
s1, calibrating the positions of an array line laser emission source and a CCD photosensitive camera, so that the CCD photosensitive camera can receive complete laser beam imaging;
s2, adjusting the height of the electrochemical cell until the lower end of the laser beam image received by the CCD photosensitive camera is incomplete, and stopping height adjustment of the electrochemical cell;
and S3, adjusting the height of the glass probe tube, acquiring laser beam imaging in real time until the upper end of the laser beam imaging is incomplete, acquiring the highest point position information of the sample, setting the jump height information of the scanning ion conductance microscope in a jump scanning mode, and scanning the surface of the sample based on the highest point position information.
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| GB0801900D0 (en) * | 2008-02-01 | 2008-03-12 | Imp Innovations Ltd | Scanning probe microscopy |
| CN102455371A (en) * | 2010-10-22 | 2012-05-16 | 国家纳米技术与工程研究院 | Device and method for evaluating malignancy degree of neuroblastoma |
| CN102455355A (en) * | 2010-10-22 | 2012-05-16 | 国家纳米技术与工程研究院 | Apparatus and method for rapidly assessing nano-material on biological security of breathing system |
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| CN105842485B (en) * | 2016-03-21 | 2019-01-18 | 西安交通大学 | Double probe scanning Ion Conductance Microscope systems and its imaging method based on balanced bridge |
| CN107462745A (en) * | 2016-06-03 | 2017-12-12 | 中国科学院沈阳自动化研究所 | A kind of apparatus and method of the quadrature amplitude scanning imagery pattern of SICM |
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| CN109142797B (en) * | 2018-09-05 | 2020-03-17 | 西安交通大学 | Droplet type scanning ion conductance microscope, probe thereof and scanning method |
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| US6373049B1 (en) * | 1999-05-03 | 2002-04-16 | Industrial Technology Research Institute | Knock mode scanning near-field optical microscope |
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