CN111505345A - Atomic force microscope probe modification method based on scanning electron microscope micro-control system - Google Patents
Atomic force microscope probe modification method based on scanning electron microscope micro-control system Download PDFInfo
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- CN111505345A CN111505345A CN202010411303.8A CN202010411303A CN111505345A CN 111505345 A CN111505345 A CN 111505345A CN 202010411303 A CN202010411303 A CN 202010411303A CN 111505345 A CN111505345 A CN 111505345A
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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
- G01Q60/24—AFM [Atomic Force Microscopy] or apparatus therefor, e.g. AFM probes
- G01Q60/38—Probes, their manufacture, or their related instrumentation, e.g. holders
Abstract
A method for modifying an atomic force microscope probe based on a scanning electron microscope micro-control system belongs to the technical field of atomic force microscope measurement and comprises the following steps: preparing nanoparticles to be modified; preparing glue needed in the modification process; preparing a probe; preparing a scanning electron microscope working cabin; modifying the probe by adopting a scanning electron microscope micro-control system; the probe was dried. The modification process is visual, and the nano particles with specific sizes and dispersion states can be selected to modify the probe to be modified, so that the state of the modified nano particles is controllable; secondly, the visual operation can clearly determine the modification position, and the nano particles are precisely modified to the needle point of the probe to be modified, so that the phenomenon of 'first modification and then observation' is avoided, and the modification efficiency of the probe is improved. The method has the advantages of simple and easy operation and high modification success rate, and the obtained probes and particles have enough strength for atomic force test.
Description
Technical Field
The invention belongs to the technical field of measurement of atomic force microscopes, and particularly relates to a method for modifying an atomic force microscope probe based on a scanning electron microscope micro-control system.
Background
The rapid development of materials and nano technology promotes the continuous widening of the application field of nano materials. The nano-particles have wide application prospects in the field of biological medicine, such as targeted drug transportation, magnetic induction thermotherapy, nuclear magnetic resonance imaging, biological detection and the like, by virtue of unique surface effect, size effect, functional groups and excellent physical properties such as magnetism, heat, electricity and the like.
The research on the interaction force between the nano particles and cells is helpful for further defining the action mechanism of the nano particles in a living body and better realizing the applications of targeting, treatment, imaging and the like. An Atomic Force Microscope (AFM) is an advanced surface sensitive technology, can describe the interaction on the molecular level in real time, enables quantitative research on the processes of cell adhesion, survival, differentiation, uptake and the like to become possible, and has wide application in the aspect of cell mechanics. By modifying the AFM probe with the nano-particles, the effects of the nano-carrier and the cell membrane can be directly measured qualitatively and quantitatively, the cellular uptake and the orientation of the nano-particles in the cells can be known, and AFM images can also be used for monitoring the dynamic change process of the cell surface.
The currently common probe modification technology comprises a coating method, an adhesive method, a spraying method, a solution deposition method and the like, and because the curvature radius of the probe tip is nano-scale, the modification process is difficult to observe, and an operation method of firstly modifying and then observing is usually adopted, the efficiency and the success rate of tip modification are low. In addition, because the surface area of the needle point is limited and the dispersibility of the nanoparticles is difficult to guarantee, the monodisperse nanoparticles cannot be modified to be accurately attached to the needle point, and the subsequent tests of imaging quality and force curve are influenced. Based on the method, the size and the dispersion state of the nano particles can be selected based on a visual scanning electron microscope micro control system, the nano particles are controlled to be accurately modified at the probe needle point, the probe modification efficiency and the success rate are greatly improved, and the accuracy and the reliability of subsequent experimental results are ensured.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides an atomic force microscope probe modification method based on a visual scanning electron microscope micro-control system, which comprises the following steps: under the visual basis of electron microscope imaging, the movable sample stage carried by the scanning electron microscope and the control cantilever are utilized, the two operation probes are used for picking up glue and nano particles by controlling the three-dimensional movement of the sample stage and the operation cantilever, and the glue and the nano particles are sequentially contacted with the needle point of the probe to be modified, so that the probe is modified. The method is simple and easy to operate, the accuracy and the high efficiency of probe modification are guaranteed, and the interaction between the nano particles and cells can be directly tested by using the probe.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for modifying an atomic force microscope probe based on a scanning electron microscope micro-control system is realized based on a scanning electron microscope working cabin and comprises the following steps:
in a first step, nanoparticles to be modified are prepared
(1) And (2) adding the nano-particles 8 into a dispersing agent at room temperature, and performing ultrasonic dispersion treatment for 5-60min to obtain nano-particle dispersion liquid, wherein each 10m L of the dispersing agent corresponds to 1-100 mu g of the nano-particles 8, and the dispersing agent is methanol, absolute ethyl alcohol, n-hexane, dichloromethane or acetone.
(2) And (3) dropwise adding the nanoparticle dispersion liquid onto a silicon wafer B14, and leaving the nanoparticles to be modified in a dispersed state after the dispersing agent is volatilized. The silicon wafer B14 with the dispersed nanoparticles was fixed on a micro-platform on the top of a support bar B11 with a conductive tape. The micro platform on the top of the supporting rod B11 can rotate clockwise or anticlockwise by 0-90 degrees, and the initial position of the micro platform is perpendicular to the supporting rod B11.
Secondly, glue 9 is dropped on another silicon wafer A13, and the silicon wafer A13 is fixed on the micro platform on the top of another support bar A10 by using conductive adhesive tape. The glue is epoxy resin glue, conductive silver glue, conductive gold glue or conductive carbon powder glue.
Third, preparing the probe
Three probes are used in the modification process, namely a manipulation probe A6, a manipulation probe B5 and a probe 7 to be modified. And the cantilevers of the three operation probes A6, the operation probe B5 and the probe 7 to be modified are respectively fixed on the rotary micro-platform at the tops of the cantilever support rod A3, the cantilever support rod B4 and the support rod C12 by adopting a conductive adhesive tape, and the needle tip parts of the three probes are ensured to be positioned outside the support rod micro-platform. The curvature radius of the needlepoints of the operation probe A6 and the operation probe B5 is smaller than that of the needlepoint of the probe 7 to be modified.
Fourthly, preparing a scanning electron microscope working cabin
The scanning electron microscope working cabin comprises a scanning electron microscope micro-control system, three probes, five supporting rods and three silicon wafers.
The scanning electron microscope micro-control system is located inside a scanning electron microscope and used for observing through a scanning electron microscope imaging system, and comprises a control system, a control cantilever 1 and a round sample table 2. The control cantilever 1 is positioned above the circular sample table 2, and the control system controls the control cantilever 1 to accurately move in the XYZ three-dimensional direction and simultaneously controls the circular sample table 2 to accurately move and rotate in the XYZ three-dimensional direction.
Two cantilever support rods are arranged on the side face of the control cantilever 1, an operation probe A6 is arranged at the top of the cantilever support rod A3, and an operation probe B5 is arranged at the top of the cantilever support rod B4. Three support rods are arranged on the circular sample table 2 at intervals, a silicon wafer B14 loaded with nano-particles 8 is fixedly arranged at the top of the middle support rod B11, a silicon wafer A13 loaded with glue is fixedly arranged at the top of the support rod A10 at one side, and a silicon wafer C15 loaded with probes 7 to be modified is fixedly arranged at the top of the support rod C12 at the other side.
The micro platform on the top of the support rod where the operation probe A6 and the operation probe B5 are located rotates clockwise by 30-60 degrees, and the micro platform on the top of the support rod where the probe 7 to be modified, the nanoparticle 8 and the glue 9 are located rotates anticlockwise by 30-60 degrees, but the clockwise rotation angle value of the micro platform on the top of the support rod where the operation probe A6 and the operation probe B5 are located is not smaller than the counterclockwise rotation angle value of the micro platform on the top of the support rod where the probe to be modified, the nanoparticle and the glue are located. The relative positions of the cantilever supporting rod A3, the cantilever supporting rod B4, the supporting rod A10, the supporting rod B11 and the supporting rod C12 are only required to be guaranteed not to block each other in the moving process.
Fifthly, modifying the probe by adopting a scanning electron microscope micro-control system, wherein the probe modification process is carried out under a scanning electron microscope imaging system, and the steps are as follows:
(1) moving the control cantilever 1 to approach to the direction of the circular sample table 2, so that the needle tip of the operation probe A6 picks up the nano-particles 8 to be modified on the silicon wafer B14; and observing and picking up the nano particles 8 with proper size and in a proper dispersion state through a scanning electron microscope imaging system for subsequent modification.
(2) Manipulation arm 1 is moved so that the tip of manipulation probe B5 sticks glue 9. And moving the control cantilever 1 to enable the needle point of the operation probe B5 to be in contact with the needle point of the probe 7 to be modified, and placing the glue 9 at the needle point of the probe 7 to be modified.
(3) And moving the control cantilever 1 again to determine a proper modification position, and enabling the needle tip of the operation probe A6 to contact the needle tip of the probe 7 to be modified, so that the magnetic nano-particles 8 are modified at the needle tip position of the probe 7 to be modified, and obtaining the modified probe.
Sixth, drying the probe
And (4) drying the modified probe obtained in the fifth step for 6-24h to obtain the atomic force microscope probe of the modified magnetic nano-particles. The drying treatment is oven drying, freeze drying, supercritical carbon dioxide drying, high temperature vacuum drying or room temperature natural drying.
The invention has the beneficial effects that: the modification process of the modification method is visual, and nanoparticles with specific sizes and in a dispersion state can be selected to modify the probe to be modified, so that the state of the modified nanoparticles is controllable; secondly, the visual operation can clearly determine the modification position, and the nano particles are precisely modified to the needle point of the probe to be modified, so that the phenomenon of 'first modification and then observation' is avoided, and the modification efficiency of the probe is improved. The method has the advantages of simple and easy operation and high modification success rate, and the obtained probes and particles have enough strength for atomic force test. The cell-magnetic nanoparticle interaction obtained by testing provides further experimental verification for researches on the uptake of the particles by cells, the adhesion between the cells and the particles, the survival capability of the cells and the like.
Drawings
Fig. 1 is a schematic view showing the placement relationship of a modification tool under a scanning electron microscope micromanipulation system.
In the figure: 1, operating a cantilever; 2, a circular sample table; 3, a cantilever support rod A; 4, a cantilever support rod B; 5 operating the probe A; 6 operating the probe B; 7, a probe to be modified; 8 a nanoparticle; 9, glue; 10 supporting the rod A; 11 supporting the rod B; 12 supporting the rod C; 13, a silicon wafer A; 14, a silicon wafer B; 15 silicon wafer C.
Detailed Description
The invention is further illustrated below with reference to specific embodiments and the accompanying drawings.
A method for modifying an atomic force microscope probe based on a scanning electron microscope micro-manipulation system comprises the following steps:
the first step is to add 50 mug magnetic nanometer particle 8 into 10m L absolute ethyl alcohol at room temperature, to carry out ultrasonic dispersion treatment for 30min to obtain magnetic nanometer particle dispersion liquid, to drop 20 mug L dispersion liquid onto a silicon slice B14 (the surface area of the silicon slice B14 is 3 mm)2) And after the absolute ethyl alcohol is naturally dried, the magnetic nano-particles 8 to be modified in a dispersed state are remained. The silicon wafer B14 with the nanoparticles 8 dispersed therein was fixed to a micro-platform on the top of the supporting bar B11 with a conductive tape.
And secondly, dropping glue 9 onto the other silicon wafer A13, wherein the glue 9 is epoxy resin glue. The silicon chip A13 was fixed to the micro-platform of another support bar A10 with conductive tape.
Thirdly, preparing an operation probe A5 and an operation probe B6, wherein the curvature radius of the operation probe A5 and the curvature radius of the operation probe B6 are both 10nm, and preparing a probe 7 to be modified, wherein the curvature radius is 50 nm. The three probe cantilevers are respectively fixed on the rotating micro-platform of the cantilever supporting rod B4, the cantilever supporting rod A3 and the supporting rod C12 by using conductive adhesive, and the needle tip parts of the three probes are ensured to be outside the micro-platform.
And fourthly, preparing a scanning electron microscope working cabin. Assembling a cantilever support rod A3 and a cantilever support rod B4 on a control cantilever 1, assembling a support rod A10, a support rod B11 and a support rod C12 on a circular sample table 2, wherein the interval between the support rod A10, the support rod B11 and the support rod C12 is 10mm, the interval between the cantilever support rod A3 and the cantilever support rod B4 is 15mm, clockwise rotating 30 degrees on the cantilever support rod B4 where an operation probe A5 and an operation probe B6 are located and a micro platform on the top of the cantilever support rod A3, and anticlockwise rotating 30 degrees on the support rod C12 where a probe 7, a nanoparticle 8 and glue 9 are located, the support rod B11 and the micro platform on the top of the support rod A10 to be modified.
And fifthly, modifying the probe by using a scanning electron microscope micro-control system, wherein the modification process is carried out under a scanning electron microscope imaging system.
(1) Moving the manipulation cantilever 1 to approach the circular sample stage 2, so that the tip of the manipulation probe A5 picks up the magnetic nanoparticles 8 to be modified in the first step.
(2) Moving the control cantilever 1 to make the needle point of the operation probe B6 pick up the glue 9 in the second step; moving control cantilever 1 to make the needle point of operation probe B6 contact with the needle point of probe 7 to be modified, and placing glue 9 at the needle point of probe 7 to be modified.
(3) And moving the control cantilever 1 again to enable the needle point of the operation probe A5 to contact the needle point of the probe 7 to be modified, and modifying the magnetic nanoparticles 8 at the needle point position of the probe 7 to be modified to obtain the modified probe.
And sixthly, placing the modified probe obtained in the fifth step in an oven for drying treatment at 80 ℃ for 24 hours to obtain the atomic force microscope probe 7 of the modified magnetic nano-particles 8.
The above-mentioned embodiments only express the embodiments of the present invention, but not should be understood as the limitation of the scope of the invention patent, it should be noted that, for those skilled in the art, many variations and modifications can be made without departing from the concept of the present invention, and these all fall into the protection scope of the present invention.
Claims (4)
1. A method for modifying an atomic force microscope probe based on a scanning electron microscope micro-control system is characterized in that the method for modifying the atomic force microscope probe is realized based on a scanning electron microscope working cabin and comprises the following steps:
in a first step, nanoparticles to be modified are prepared
(1) At room temperature, adding the nano particles (8) into a dispersing agent, and performing ultrasonic dispersion to obtain a nano particle dispersion liquid;
(2) dripping the nano-particle dispersion liquid on a silicon wafer B (14), and leaving the nano-particles to be modified in a dispersed state after the dispersing agent is volatilized; fixing the silicon chip B (14) dispersed with the nano particles on a micro platform at the top of the support rod B (11) by using a conductive adhesive tape; the micro platform at the top of the supporting rod B (11) can rotate clockwise or anticlockwise by 0-90 degrees, and the initial position of the micro platform is vertical to the supporting rod B (11);
secondly, dripping glue (9) on the other silicon wafer A (13), and fixing the silicon wafer A (13) on a micro platform at the top of the other supporting rod A (10) by using a conductive adhesive tape;
third, preparing the probe
Three probes are used in the modification process, namely an operation probe A (6), an operation probe B (5) and a probe to be modified (7); fixing cantilevers of the three operation probes A (6), the operation probe B (5) and the probe (7) to be modified on a rotary micro-platform at the top of a cantilever support rod A (3), a cantilever support rod B (4) and a support rod C (12) respectively by adopting a conductive adhesive tape, and ensuring that the tip parts of the three probes are positioned outside the support rod micro-platform; the curvature radius of the needle points of the operation probe A (6) and the operation probe B (5) is smaller than that of the needle point of the probe (7) to be modified;
fourthly, preparing a scanning electron microscope working cabin
The scanning electron microscope working cabin comprises a scanning electron microscope micro-control system, three probes, five support rods and three silicon wafers;
the scanning electron microscope micro-control system is positioned inside a scanning electron microscope and is used for observing through a scanning electron microscope imaging system, and comprises a control system, a control cantilever (1) and a round sample table (2); the control cantilever (1) is positioned above the circular sample table (2), and the control system controls the control cantilever (1) to accurately move in the XYZ three-dimensional direction and simultaneously controls the circular sample table (2) to accurately move and rotate in the XYZ three-dimensional direction;
two cantilever support rods are arranged on the side face of the control cantilever (1), an operation probe A (6) is arranged at the top of the cantilever support rod A (3), and an operation probe B (5) is arranged at the top of the cantilever support rod B (4); three support rods are arranged on the circular sample table (2) at intervals, a silicon wafer B (14) loaded with nano particles (8) is fixedly arranged at the top of a middle support rod B (11), a silicon wafer A (13) loaded with glue is fixedly arranged at the top of a support rod A (10) at one side, and a silicon wafer C (15) loaded with probes (7) to be modified is fixedly arranged at the top of a support rod C (12) at the other side;
the micro platform at the top of the supporting rod where the operation probe A (6) and the operation probe B (5) are located rotates clockwise by 30-60 degrees, the micro platform at the top of the supporting rod where the probe (7), the nano particles (8) and the glue (9) to be modified rotate anticlockwise by 30-60 degrees, and the clockwise rotation angle value of the micro platform at the top of the supporting rod where the operation probe A (6) and the operation probe B (5) are located is not smaller than the anticlockwise rotation angle value of the micro platform at the top of the supporting rod where the probe, the nano particles and the glue to be modified are located; the relative positions of the cantilever support rod A (3), the cantilever support rod B (4), the support rod A (10), the support rod B (11) and the support rod C (12) are only required to be ensured not to block each other in the moving process;
fifthly, modifying the probe by adopting a scanning electron microscope micro-control system, wherein the probe modification process is carried out under a scanning electron microscope imaging system, and the steps are as follows:
(1) moving the control cantilever (1) to approach the circular sample table (2) so that the needle point of the operation probe A (6) picks up the nano particles (8) to be modified on the silicon wafer B (14); and observing and picking up the nano particles (8) with proper size and in a proper dispersion state through a scanning electron microscope imaging system;
(2) moving the control cantilever (1) to enable the needle point of the operation probe B (5) to stick glue (9); moving the control cantilever (1) to enable the needle point of the operation probe B (5) to be in contact with the needle point of the probe (7) to be modified, and placing the glue (9) at the needle point of the probe (7) to be modified;
(3) moving the control cantilever (1) again to determine a proper modification position, and enabling the needle point of the operation probe A (6) to contact the needle point of the probe (7) to be modified, so that the magnetic nanoparticles (8) are modified at the needle point position of the probe (7) to be modified, and a modified probe is obtained;
sixth, drying the probe
And drying the modified probe obtained in the fifth step to obtain the atomic force microscope probe of the modified magnetic nano-particles.
2. The atomic force microscope probe modification method based on the scanning electron microscope micro-manipulation system is characterized in that in the first step, 1-100 μ g of nanoparticles (8) correspond to every 10m L of dispersing agents, wherein the dispersing agents are methanol, absolute ethyl alcohol, n-hexane, dichloromethane or acetone.
3. The atomic force microscope probe modification method based on the scanning electron microscope micro-manipulation system, according to claim 1, characterized in that the glue is epoxy glue, conductive silver glue, conductive gold glue or conductive carbon powder glue.
4. The atomic force microscope probe modification method based on the scanning electron microscope micro-manipulation system according to claim 1, wherein the drying treatment is oven drying, freeze drying, supercritical carbon dioxide drying, high-temperature vacuum drying or room-temperature natural drying.
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