Scanning ionic conductance microprobe system and scanning method thereof

Abstract

The invention discloses a scanning ionic conductance microprobe system and a scanning method thereof. The scanning ionic conductance microprobe system comprises a motion control unit, a probe, a signal acquisition unit, a controller and a TMR sensor, the probe is installed on the motion control unit, the motion control unit is connected with the controller, the TMR sensor is arranged at the bottom of the probe, the output end of the signal acquisition unit is connected with the controller, the input end of the signal acquisition unit is connected with the probe, the TMR sensor and the substrate, and a magnetic layer is arranged on the probe. By integrating the technologies of high-precision positioning, multi-signal acquisition, magnetic induction and the like, the accuracy and efficiency of living cell detection are effectively improved, multi-dimensional information of living cells can be accurately obtained in real time, technical support is provided for specific living cell detection and drug pathology analysis, a powerful research tool is provided for scientific research personnel, and the method is suitable for popularization and application. Wide application prospects are realized.

Description

Technical Field

[0001] The invention belongs to the technical field of microscope application, in particular to a scanning ion conductivity microprobe system and a scanning method thereof. Background Art

[0002] Cells are the basic units of life and form the cornerstone of all known organisms on Earth. From single-celled organisms to complex human organisms, every life form is composed of a large number of cells of various types, each with its own structure and function, which together maintain the normal functioning of the organism. Cytology, as a basic discipline, has always occupied a pivotal position in the vast field of biological sciences.

[0003] Cytological research covers all levels of cells. It takes cells as the research object and studies the structure and function of cells and organelles, the life history of cells and the laws of various life activities at three levels: the overall level of cells, the submicroscopic level, and the molecular level. It promotes the continuous in-depth research and development of genetics and development, and also provides an important scientific basis for understanding the nature and mechanism of life.

[0004] With the advancement of science and technology, the application of microscopic tools, especially probe microscopy, has become increasingly widespread. The susceptible, label-free, in situ and real-time detection methods of living cells have gradually gained attention in the biomedical field, showing the research advantages of long-term and dynamic monitoring of living cells under physiological and pathological conditions. Scanning ion conductance microscopy (SICM) is one of the best tools for observing living cells under physiological conditions today. It can not only perform nanoscale imaging of cell morphology and sub-organs, but also continuously monitor ion channels on the cell membrane surface, and analyze the biological characteristics of cell metabolism. SICM realizes height detection based on ion current. When the distance between the tip of its probe and the morphology is less than the opening diameter, the ion current mutates, and the current scanning height is calculated by capturing the mutated ion current. However, in the actual cell scanning process, due to the use of jump mode scanning, the point scanning needs to repeatedly jump to a safe height to explore and scan downward, resulting in redundant scanning paths, slow scanning speed, and low scanning efficiency. At the same time, the sensitivity of the scanning probe to detect the current determines the accuracy of the detection height and the return time to a certain extent, which in turn affects the performance and scanning results of SICM scanning. Therefore, improving the scanning probe and optimizing the scanning method have always been urgent issues to be solved in SICM fast scanning. Summary of the invention

[0005] The object of the present invention is to provide a scanning ion conductivity microprobe system and a scanning method thereof, so as to solve the problems of low scanning accuracy and efficiency in the prior art.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions: A scanning ion conductivity microprobe system, comprising a motion control unit, a probe, a signal acquisition unit, a controller and a TMR sensor; The probe is installed on a motion control unit, the motion control unit is connected to a controller, the TMR sensor is arranged at the bottom of the probe, the output end of the signal acquisition unit is connected to the controller, the input end of the signal acquisition unit is respectively connected to the probe, the TMR sensor and the substrate, and a magnetic layer is arranged on the probe.

[0007] Furthermore, the motor in the motion control unit is connected to the piezoelectric ceramic motion module, the piezoelectric ceramic motion module is connected to the probe, the motor is connected to the controller, and the piezoelectric ceramic motion module is a three-dimensional motion structure.

[0008] Furthermore, the probe also includes a probe layer and a separation layer, the probe layer is a glass microprobe, the separation layer is arranged on the surface of the probe layer, the magnetic layer is arranged on the surface of the separation layer, and the magnetic layer is a ferrosoferric oxide annular coating.

[0009] Furthermore, a current amplifier is provided on the ion current circuit between the input end of the signal acquisition unit and the probe, and a host computer is installed on the ion current circuit between the output end of the signal acquisition unit and the controller.

[0010] Furthermore, the input end of the signal acquisition unit is respectively connected to the TMR sensor and the substrate through an ion current circuit.

[0011] Furthermore, the controller is connected to the motor in the motion control unit via an ion current circuit.

[0012] Furthermore, the substrate is arranged between the probe and the TMR sensor, and a substrate solution composed of a solution and cells to be detected is arranged in the substrate.

[0013] A scanning method of the scanning ion conductivity microprobe system, comprising: Turn on the TMR sensor, which continuously performs magnetic detection. At the same time, the probe is driven by the piezoelectric ceramic motion module in the motion control unit and moves downward along the Z axis from a safe height. When the value of the TMR sensor collected by the signal acquisition unit exceeds the low value, the controller controls the probe to slow down and continue to move downward; When the value of the TMR sensor collected by the signal acquisition unit exceeds the high value, the controller controls the probe to slow down and continue to move downward; Turn off the TMR sensor, and the controller controls the probe to slow down and continue to approach the substrate. When the signal acquisition unit collects the sudden ion current, it records the current height value and scans and collects information about the cells to be tested in the substrate; The controller controls the probe to return to a safe height, and the piezoelectric ceramic motion module in the motion control unit drives the probe to move along the X and Y directions to the next point to be scanned, and repeats the above process until the final point of the set scanning size is scanned.

[0014] Furthermore, the low value and the high value are both magnetic detection values, which are used to calibrate the starting point of the deceleration adjustment.

[0015] Furthermore, the speed of the probe moving downward along the Z axis is controlled by the following function:

[0016] in, represents the initial motion speed of the probe, Indicates the movement speed of the probe after deceleration.

[0017] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a scanning ion conductivity microprobe system, by installing the probe on a motion control unit, the motion control unit is connected to a controller, and the controller can control the motion control unit to realize high-precision positioning and movement of the probe in three-dimensional space. The probe is precisely manipulated on a microscopic scale to ensure the accuracy and reliability of the experimental results. The output end of the signal acquisition unit is connected to the controller, and the input end of the signal acquisition unit is respectively connected to the probe, the TMR sensor and the substrate, and multiple signals can be simultaneously collected and output to the controller. The multi-parameter detection method provides rich sample information, and realizes real-time display, storage and further processing of data. By comprehensively analyzing these signals, a more in-depth and accurate understanding of the sample to be tested can be achieved. The TMR sensor is arranged at the bottom of the probe, and the probe has a magnetic layer, which can use the magnetic effect to sense the proximity of the morphological position, and collect multi-dimensional information of cell subcellular characteristics at the nanoscale, and carry out targeted acquisition and analysis through the characteristics of different information, which can more accurately identify and analyze the fine structure and function of the cell, and provide a powerful tool for cell biology research. The present invention effectively improves the accuracy and efficiency of living cell detection by integrating high-precision positioning, multi-signal acquisition, and magnetic induction technologies. It can obtain multi-dimensional information of living cells in real time and accurately, and provides technical support for specific living cell detection and drug pathology analysis. It provides a powerful research tool for scientific researchers and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0019] Figure 1 It is a schematic diagram of the structure of the scanning ion conductivity microprobe system of the present invention.

[0020] Figure 2 It is a schematic diagram of the structure of the scanning ion conductivity microprobe of the present invention.

[0021] Wherein: 1-motion control unit, 2-probe, 3-current amplifier, 4-signal acquisition unit, 5-host computer, 6-controller, 7-TMR sensor, 8-substrate, 9-ion current circuit, 21-probe layer, 22-magnetic layer, 23-separation layer. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Generally, the components of the embodiments of the present invention described and shown in the drawings here can be arranged and designed in various different configurations.

[0023] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0024] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0025] In the description of the embodiments of the present invention, it should be noted that if the terms "upper", "lower", "horizontal", "inner", etc. indicate an orientation or positional relationship based on the orientation or positional relationship shown in the drawings, or the orientation or positional relationship in which the product of the invention is usually placed when in use, it is only for the convenience of describing the present invention and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. In addition, the terms "first", "second", etc. are only used to distinguish the description, and cannot be understood as indicating or implying relative importance.

[0026] In addition, if the term "horizontal" appears, it does not mean that the component must be absolutely horizontal, but can be slightly tilted. For example, "horizontal" only means that its direction is more horizontal than "vertical", which does not mean that the structure must be completely horizontal, but can be slightly tilted.

[0027] In the description of the embodiments of the present invention, it is also necessary to explain that, unless otherwise clearly specified and limited, the terms "set", "install", "connect", and "connect" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal connection of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0028] The present invention is further described in detail below in conjunction with the accompanying drawings: See also Figure 1 The present invention provides a scanning ion conductivity microprobe system, comprising a motion control unit 1, a probe 2, a current amplifier 3, a signal acquisition unit 4, a host computer 5, a controller 6, a TMR sensor 7, a substrate 8 and an ion current circuit 9. The probe 2 is mounted on the motion control unit 1, the motion control unit 1 is connected to the controller 6, and the controller 6 controls the motion control unit 1 to drive the probe 2 to move.

[0029] The motion control unit 1 is the core driving part of the whole system, including a motor and a piezoelectric ceramic motion module. The motor is used as a power source and is connected to the piezoelectric ceramic motion module. The piezoelectric ceramic motion module is connected to the probe 2 to ensure the high stability and accuracy of the motion of the probe 2. The motor is connected to the controller 6 through the ion current circuit 9. The piezoelectric ceramic motion module is a three-dimensional motion structure with three-dimensional motion capabilities in the X, Y, and Z directions. The piezoelectric ceramic motion module can achieve nanometer-level precise displacement control and can perform fine positioning and movement in three-dimensional space.

[0030] like Figure 2As shown, the probe 2 includes a probe layer 21, a separation layer 22 and a magnetic layer 23. The probe layer 21 is an original drawn glass microprobe, and its surface has a certain roughness; the separation layer 22 is densely arranged by sputtering using insulating materials, which separates the solution in the probe from the magnetic layer 23, thereby preventing unnecessary electrochemical interference and magnetic interference, and ensuring the accuracy and reliability of the experiment; the magnetic layer 23 is an annular coating containing the magnetic substance ferrosoferric oxide, which not only enhances the magnetic response ability of the probe, but also closely adheres to the probe surface by sputtering, thereby improving the stability and durability of the overall structure.

[0031] The signal acquisition unit 4 is used to collect the ion current signal in the ion current circuit 9, and the ion current circuit 9 is a current loop composed of a probe 2, a substrate 8, a signal acquisition unit 4, and a current amplifier 3, which realizes the accurate measurement and transmission of the ion current. The output end of the signal acquisition unit 4 is connected to the controller 6 through the ion current circuit 9, and the input end of the signal acquisition unit 4 is respectively connected to the probe 2, the TMR sensor 7 and the substrate 8 through the ion current circuit 9. The ion current circuit 9 between the input end of the signal acquisition unit 4 and the probe 2 is provided with a current amplifier 3, and the current amplifier 3 is used to amplify the ion current into a detectable current, which greatly improves the intensity of the ion current signal, so that it can be accurately captured by the signal acquisition unit 4 and converted into analyzable data. The host computer 5 is installed on the ion current circuit 9 between the output end of the signal acquisition unit 4 and the controller 6. The host computer 5 is a scanning system interactive interface, which not only provides an intuitive and easy-to-use operating interface, but also realizes functions such as motion control, data display and interaction, and communication control. The user can easily set the parameters such as the motion trajectory and speed of the probe through the host computer 5, monitor the ion current changes during the experiment in real time, and communicate and remotely control data with other devices. The TMR sensor 7 is disposed at the bottom of the probe 2. The TMR sensor 7 is a highly sensitive magnetic detector that can accurately detect magnetic changes in the probe 2, providing an additional detection dimension for the experiment. The substrate 8 is disposed between the probe 2 and the TMR sensor 7. The substrate 8 is a substrate solution composed of a solution and cells to be tested.

[0032] The present invention also provides a scanning method of a scanning ion conductivity microprobe system, which specifically comprises the following steps: (1) Turn on the TMR sensor 7, which continuously performs magnetic detection. At the same time, the probe 2 is driven by the piezoelectric ceramic motion module in the motion control unit 1 to move rapidly downward along the Z axis from a set safety height; (2) When the value of the TMR sensor 7 collected by the signal acquisition unit 4 exceeds the set low value, the controller 6 controls the probe 2 to decelerate and continue to move downward at a low speed; (3) When the value of the TMR sensor 7 collected by the signal acquisition unit 4 exceeds the set high value, the controller 6 controls the probe 2 to decelerate and continue to move downward at a slow speed; (4) Turn off the TMR sensor 7, and the controller 6 controls the probe 2 to slow down and then approach the substrate 8 at an extremely slow speed. When the signal acquisition unit 4 detects a sudden ion current in the ion current circuit 9, the current height value is recorded, and information scanning and collection of the cells to be tested in the substrate 8 are performed.

[0033] (5) The controller 6 controls the probe 2 to quickly return to the set safety height, and the piezoelectric ceramic motion module in the motion control unit (1) drives the probe 2 to move along the X and Y directions to the next point to be scanned.

[0034] (6) Repeat steps (1) to (5) until the scan reaches the final point of the set scan size.

[0035] The set safety height is a starting point for the downward movement of the probe 2, and is used to prevent the probe 2 from colliding during movement.

[0036] The set low value is the magnetic detection value of the TMR sensor 7 at a low position, which is used to calibrate the starting point of low speed adjustment after setting; the set high value point is the magnetic detection value of the TMR sensor 7 at a high position, which is used to calibrate the starting point of slow speed adjustment after setting.

[0037] When the probe 2 moves downward along the Z axis, the fast speed is the set initial movement speed, and the low speed, slow speed, and very slow speed are the speeds at which the probe 2 moves downward along the Z axis, which are controlled by the following function:

[0038] in, represents the initial motion speed of the probe, Indicates the speed corresponding to the low speed, slow speed, and extremely slow speed of the probe after deceleration.

[0039] The present invention is further described in detail below in conjunction with specific embodiments: Embodiment 1: In this embodiment, the scanning method of the scanning ion conductivity microprobe system includes a preparation stage and a scanning stage.

[0040] The preparation stage mainly involves TMR magnetic value calibration and placement of probe 2 and substrate 8. Indicates that the TMR magnetic value calibration is the maximum magnetic value of the probe 2 when it moves along the Z axis using the TMR sensor 7. and minimum magnetic value The probe 2 and the substrate 8 are placed by installing the probe 2 on the piezoelectric ceramic motion module and fixing it, and placing the cell sample to be tested on the substrate 8. In the preparation stage, the piezoelectric ceramic motion module drives the probe 2 to move from top to bottom along the Z axis, and the host computer 5 reads the corresponding maximum magnetic value. and minimum magnetic value .

[0041] During the scanning phase, set the safe height , set the initial movement speed , set the low value to , set the high value to , turn on the TMR sensor 7, and control the piezoelectric ceramic motion module through the controller 6 to drive the probe 2 from the set safety height to Move downward along the Z axis, while the TMR sensor 7 continuously performs magnetic detection. When the detected magnetic value is greater than When the speed adjustment is turned on, the current speed remains unchanged; when it exceeds the set low value , with numerical value Continue to move downward at a low speed; when it exceeds the set high value , with numerical value Then the TMR sensor 7 is turned off and the value is The probe 2 moves downward at a very slow speed. When the signal acquisition unit 4 detects a sudden ion current in the ion current circuit 9, it records the current height value and scans and collects information about the cells to be tested in the substrate 8. The controller 6 controls the motion control unit 1 to drive the probe 2 to return to the set safe height. The piezoelectric ceramic motion module is used to drive the probe 2 to move along the X and Y directions to the next point to be scanned. This process is repeated until the scanning is completed.

[0042] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A scanning ion conductivity microprobe system, characterized in that: It comprises a motion control unit (1), a probe (2), a signal acquisition unit (4), a controller (6) and a TMR sensor (7); The probe (2) is mounted on a motion control unit (1), the motion control unit (1) is connected to a controller (6), the TMR sensor (7) is arranged at the bottom of the probe (2), the output end of the signal acquisition unit (4) is connected to the controller (6), the input end of the signal acquisition unit (4) is respectively connected to the probe (2), the TMR sensor (7) and the substrate (8), and a magnetic layer (23) is arranged on the probe (2).

2. A scanning ion conductivity microprobe system according to claim 1, characterized in that: The motor in the motion control unit (1) is connected to a piezoelectric ceramic motion module, the piezoelectric ceramic motion module is connected to a probe (2), the motor is connected to a controller (6), and the piezoelectric ceramic motion module is a three-dimensional motion structure.

3. A scanning ion conductivity microprobe system according to claim 1, characterized in that: The probe (2) further comprises a probe layer (21) and a separation layer (22); the probe layer (21) is a glass microprobe; the separation layer (22) is arranged on the surface of the probe layer (21); the magnetic layer (23) is arranged on the surface of the separation layer (22); and the magnetic layer (23) is a ferrosoferric oxide annular coating.

4. A scanning ion conductivity microprobe system according to claim 1, characterized in that: A current amplifier (3) is provided on the ion current circuit (9) between the input end of the signal acquisition unit (4) and the probe (2), and a host computer (5) is installed on the ion current circuit (9) between the output end of the signal acquisition unit (4) and the controller (6).

5. A scanning ion conductivity microprobe system according to claim 1, characterized in that: The input end of the signal acquisition unit (4) is respectively connected to the TMR sensor (7) and the substrate (8) through an ion current circuit (9).

6. A scanning ion conductivity microprobe system according to claim 1, characterized in that: The controller (6) is connected to the motor in the motion control unit (1) via an ion current circuit (9).

7. A scanning ion conductivity microprobe system according to claim 1, characterized in that: The substrate (8) is arranged between the probe (2) and the TMR sensor (7), and a substrate solution composed of a solution and cells to be detected is arranged in the substrate (8).

8. A scanning method of the scanning ion conductivity microprobe system according to any one of claims 1 to 7, characterized in that: include: The TMR sensor (7) is turned on, and the TMR sensor (7) continuously performs magnetic detection, while the probe (2) is driven by the piezoelectric ceramic motion module in the motion control unit (1) to move downward along the Z axis from a safe height; When the value of the TMR sensor (7) collected by the signal collection unit (4) exceeds the lower value, the controller (6) controls the probe (2) to slow down and continue to move downward; When the value of the TMR sensor (7) collected by the signal collection unit (4) exceeds the high value, the controller (6) controls the probe (2) to slow down and continue to move downward; The TMR sensor (7) is turned off, and the controller (6) controls the probe (2) to slow down and continue to approach the substrate (8). When the signal acquisition unit (4) acquires the mutant ion current, the current height value is recorded, and information scanning and acquisition of the cells to be tested in the substrate (8) are performed; The controller (6) controls the probe (2) to return to a safe height, and the piezoelectric ceramic motion module in the motion control unit (1) drives the probe (2) to move along the X and Y directions to the next point to be scanned, and the above process is repeated until the final point of the set scanning size is scanned.

9. The scanning method of the scanning ion conductivity microprobe system according to claim 8, characterized in that: The low value and the high value are both magnetic detection values, which are used to calibrate the starting point of the deceleration adjustment.

10. The scanning method of the scanning ion conductivity microprobe system according to claim 8, characterized in that: The speed of the probe (2) moving downward along the Z axis is controlled by the following function: in, represents the initial motion speed of the probe, Indicates the movement speed of the probe after deceleration.

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