CN111505346A - AFM probe for quantitative measurement, modification method and application thereof - Google Patents
AFM probe for quantitative measurement, modification method and application thereof Download PDFInfo
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- CN111505346A CN111505346A CN202010411793.1A CN202010411793A CN111505346A CN 111505346 A CN111505346 A CN 111505346A CN 202010411793 A CN202010411793 A CN 202010411793A CN 111505346 A CN111505346 A CN 111505346A
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
- A61N2/00—Magnetotherapy
- A61N2/004—Magnetotherapy specially adapted for a specific therapy
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
- A61N2/00—Magnetotherapy
- A61N2/06—Magnetotherapy using magnetic fields produced by permanent magnets
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
Abstract
The invention provides an AFM probe for quantitative measurement, a modification method and application thereof, and belongs to the technical field of nano mechanics and experimental mechanics. The invention uses micron-sized small balls with defects on the surfaces after special treatment to uniformly adsorb a layer of nano particles, and then the micron-sized small balls are attached to the point of the AFM probe to finish probe modification, and the method specifically comprises the following steps: pretreating the micron-sized pellets; preparing a nanoparticle dispersion liquid; the micron-sized globules absorb the nano-particles; selecting magnetic nanoparticle modified micron-sized spheres; AFM probe modification was performed. The method is simple and convenient to operate and wide in application range, and the problem that the agglomeration shape of the nano particles at the tip of the AFM probe is uncontrollable in conventional modification is solved; and the interaction force between the unit nano particles and the surface of the measured object is obtained through observation and calculation of a scanning electron microscope, so that quantitative nano mechanical measurement is realized.
Description
Technical Field
The invention belongs to the technical field of nano mechanics and experimental mechanics, relates to atomic force microscope probe modification, and particularly relates to an AFM probe for quantitative measurement, a modification method and application thereof.
Background
The nano material has unique microstructure and tissue morphology, which causes great changes in physical, chemical and mechanical properties, thereby greatly widening the application field of the material. With the development of nanotechnology, on one hand, the application of Engineered Nanoparticles (ENPs) is more and more extensive, which inevitably causes the human body to be exposed in the nano environment, causing potential adverse effects on the body; on the other hand, the nano-particles, as a novel material, also play a great role in the field of biological medicine, such as targeted delivery of drugs, magnetic hyperthermia, gene therapy, and the like. In order to develop an effective and reliable prediction method and a protective measure to determine the absorption condition of the cell to the nanoparticle, the interaction force between the cell and the nanoparticle needs to be qualitatively and quantitatively determined, and reliable reference data is provided for experimental verification and numerical simulation. Atomic Force Microscopy (AFM), one of the most advanced surface-sensitive technologies today, is capable of characterizing interaction forces in real time at the molecular level. The development of the AFM probe (atomic force microscope probe) tip modification technology greatly widens the application range, and makes qualitative and quantitative determination of interaction forces of various surfaces, molecules, groups and the like easier. The AFM probe tip modified by the nano particles can be used for carrying out experimental operation on living cells and measuring the interaction force between the nano particles and the cells in real time.
The nano particles are extremely small in size and easy to agglomerate, single nano particle control is difficult to realize, a large number of nano particles are mostly treated as objects, various methods for modifying the AFM probe tip by using the nano particles exist at present, most of the methods are directly used for modifying the AFM probe tip, the nano particles are dispersed into a solution, the probe tip is contacted with the solution, the nano particles are adsorbed on the surface of the probe tip, the dispersion can wet the whole probe tip or even a cantilever part due to the action of capillary force, if the cantilever is polluted by the particles, the reflection of the cantilever to laser is poor, the measurement precision is influenced, the modification effect is limited by the dispersion degree of the nano particle dispersion, the modified AFM probe is irregular due to the existence of agglomeration of the nano particles with uncontrollable shapes, the distribution of the tip nano particles is irregular, and even if the press-in depth and the adhesion force are obtained, in addition, the method is unpredictable to a great extent, the common observation method cannot see the nanoparticles in the modification process, the modification effect can be judged only by observing the probe through a scanning electron microscope after modification, and the consumption of the probe is increased due to the uncertainty of the modification, so that the time and the economic cost of the experiment are increased.
Disclosure of Invention
In order to solve the problems, the invention provides a method for modifying an AFM probe by using shape-controllable nano particles.
The technical scheme of the invention is as follows:
an AFM probe for quantitative measurement, the probe has the following specific structure: micron-sized balls are adhered to the tip of the AFM probe, a large number of amorphous concave-convex structures are irregularly distributed on the surfaces of the micron-sized balls to form rough surfaces, and nano particles are uniformly adhered to the rough surfaces of the balls. The roughness of the rough surface is in the nanometer level, and the concave-convex peak-valley value of the rough surface is smaller than the size of the nano particles to be modified.
An AFM probe modification method for quantitative measurement comprises the following steps:
step 1, pretreatment of micron-sized pellets
(1) The micron-sized globules are pretreated by a chemical etching or mechanical processing method to form a rough surface microstructure which can increase the adsorption effect of the micron-sized globules on the nano-particles.
The etching time of the chemical etching is 0.05-2h, the etching time is determined according to different pellet materials and selected corrosive liquid, and the chemical etching adopts a conventional acid washing etching method. The mechanical processing method adopts a ball milling method of a ball mill, the ball milling time is between 0.05 and 10 hours, and the ball milling time is determined according to different small ball materials and ball milling modes.
The micron-sized balls are made of carbon, titanium, silicon dioxide, polystyrene and the like, the micron-sized balls are spherical, the diameters of the micron-sized balls are adjusted according to the type of the AFM probe, the shape and the diameter of the nano particles, the spring coefficient of an AFM probe cantilever is influenced by the weight of the micron-sized balls when the diameters of the micron-sized balls are too large, the nano particles are unevenly distributed on the surface of the AFM probe cantilever to be increased when the diameters of the micron-sized balls are too small, and the diameters of the micron-sized balls are 0.
(2) Carrying out ultrasonic treatment on the pretreated micron-sized pellets for 0.2-2h, filtering the ultrasonic solution by a filter screen with the diameter equivalent to that of the pellets for 3-10 times to obtain clean micron-sized pellets, and then placing the micron-sized pellets in an oven for drying, wherein the temperature can be selected from 40-120 ℃ according to the material of the pellets, and the drying time can be 1-24h according to the temperature.
Step 2, preparing nanoparticle dispersion liquid
Adding the nano particles into a dispersing agent, and carrying out ultrasonic dispersion treatment on the nano particles to obtain nano particle dispersion liquid, wherein the concentration of the nano particle dispersion liquid is 0.1-1mg/ml, and the nano particle dispersion liquid is adjusted according to different nano particle types.
The dispersing agent is deionized water, absolute ethyl alcohol and the like, and is selected on the premise of not polluting nano particles and micron-sized balls.
The nanoparticles can be conventional various functional nanoparticles, such as magnetic nanoparticles and the like. The size of the nano particles is between 10 and 80 nm.
Step 3, adsorbing the nano particles by the micron-sized beads
And (3) adding the micron-sized balls prepared in the step (1) into the nano-particle dispersion liquid, wherein the concentration of the micron-sized balls is 0.01-0.1mg/ml, performing ultrasonic treatment for 0.2-2h again to obtain a mixed dispersion liquid, and fully contacting the nano-particles in the nano-particle dispersion liquid with the micron-sized balls so as to adsorb the nano-particles on the surfaces of the micron-sized balls. Dropping the mixed dispersion liquid in the center of a silicon chip or a mica sheet, drying, and placing in an oven for drying, wherein the temperature can be selected from 40-200 ℃ according to the material of the pellets and the type of the nano particles, and the drying time can be 1-48h according to the temperature.
Step 4, AFM probe modification
At the moment, the nano particles are attached to the surfaces of the micron-sized spheres, the sizes of the micron-sized spheres become clearly visible under an optical microscope, and the micron-sized spheres are attached to the tips of AFM probes through a micromanipulator. In order to make the adsorption firm, the adhesion operation is performed by means of gluing or the like.
The AFM probe can be used for measuring the interaction force between unit nanoparticles and the surface of a measured object, provides an important tool for researching the adsorption of the nanoparticles and other objects, can directly obtain key parameters such as the adhesion force of the nanoparticles and other observed objects, for example, the adhesion force research of magnetic nanoparticles and cancer cells, and can select proper magnetic nanoparticles from the adhesion force to carry out magnetic thermotherapy on the cancer cells. The method comprises the following specific steps:
the AFM probe prepared by the invention can be observed by a scanning electron microscope, the same area is selected at different positions of a micron-sized bead at the tip of the AFM probe to count nanoparticles, the density of the nanoparticles at different positions is calculated, and the average value is taken as the density rho of the nanoparticles on the surface of the micron-sized bead; and measuring the integral radius R of the micron-sized balls with the surfaces modified with the nano particles. And then placing the prepared AFM probe on a probe head of an atomic force microscope, controlling the z-axis displacement of the atomic force microscope to enable the nano particles on the micron-scale small ball surface of the AFM probe point to be in contact with the measured object and to be pressed into the measured object for a certain depth d, and then separating from the surface of the measured object. And recording the interaction force F (adhesive force) between the nano particles on the surface of the micron-sized sphere of the AFM probe tip and the surface of the measured object when the AFM probe tip is separated. Obtaining the contact area S of the nano particles and the object to be measured as 2 pi Rd according to the press-in depth d, multiplying the contact area S by the density rho of the nano particles on the surface of the micron-sized small spheres to obtain the number N of the nano particles participating in interaction, and dividing the interaction force F (adhesive force) measured by an experiment by the number N of the nano particles to obtain the adhesive force between the unit nano particles and the surface of the object to be measured. Provides an important tool for researching the adsorption of the nano particles and other objects. For example, the adhesion force research of the magnetic nanoparticles and the cancer cells, and the magnetic thermotherapy on the cancer cells can be performed by selecting the appropriate magnetic nanoparticles from the adhesion force.
The invention has the beneficial effects that: the method has simple and convenient operation steps and wide application range, and avoids the problem that the agglomeration shape of the nano particles is uncontrollable because the nano particles are directly modified at the tip of the AFM probe in the conventional modification. The probe prepared by the invention can conveniently measure the integral radius of the micron-sized small ball with the surface modified with the nano particles by a scanning electron microscope observation method, simultaneously calculate the density of the nano particles on the surface of the micron-sized small ball, and measure the interaction force (adhesive force) between the unit nano particles and the surface of a measured object by an experiment, thereby realizing quantitative nano mechanical measurement and providing important tools and key parameters for researching the adsorption of the nano particles and other objects.
Drawings
Fig. 1 is a diagram of a surface-treated microsphere.
Fig. 2 is a schematic of nanoparticle adsorption.
FIG. 3 is a schematic diagram of probe modification.
Detailed Description
The following detailed description of the invention refers to the accompanying drawings.
An AFM probe for quantitative measurement, the probe has the following specific structure: micron-sized balls are adhered to the tip of the AFM probe, a large number of amorphous concave-convex structures are irregularly distributed on the surfaces of the micron-sized balls to form rough surfaces, and nano particles are uniformly adhered to the rough surfaces of the balls. The roughness of the rough surface is in the nanometer level, and the concave-convex peak-valley value of the rough surface is smaller than the size of the nano particles to be modified.
An AFM probe modification method for quantitative measurement enables a tip of an AFM probe to be modified with a carbon microsphere with a surface uniformly adsorbing magnetic nanoparticles, and comprises the following steps:
step 1, pretreatment of carbon microspheres
(1) Putting carbon microspheres with the diameter of 0.5 mu m into a ball mill for pretreatment, wherein the rotating speed is 300r/min and the time is 5 min; then carrying out acid washing treatment on the carbon microspheres for 5 min;
(2) taking out the carbon microspheres, ultrasonically cleaning the carbon microspheres for 15min by using deionized water, repeatedly cleaning and filtering for three times, cleaning and filtering residues on the surfaces of the carbon microspheres, and drying the carbon microspheres for 10h at 80 ℃;
step 2, preparing nanoparticle dispersion liquid
And (3) taking magnetic nanoparticles with the diameter of 40nm, adding absolute ethyl alcohol to disperse the magnetic nanoparticles with the concentration of 0.1mg/ml, and performing ultrasonic dispersion for 15min to achieve the optimal dispersion effect.
Step 3, adsorbing the nano particles by the carbon microspheres
And (3) taking a proper amount of carbon microspheres, adding the carbon microspheres into the magnetic nanoparticle dispersion liquid subjected to ultrasonic dispersion in the step (2), and performing ultrasonic treatment for 30min again to obtain a mixed dispersion liquid so as to enable the nanoparticles in the magnetic nanoparticle dispersion liquid to be in full contact with the carbon microspheres. Dropping the mixed dispersion liquid in the center of a silicon chip, drying the silicon chip, and drying the silicon chip in an oven at 80 ℃ for 10 hours.
Step 4, AFM probe modification
And (3) adhering the carbon microspheres obtained in the step (3) to the tip of the AFM probe by using a micromanipulator.
The AFM probe is placed on a probe head of an atomic force microscope, nano particles on the surface of a carbon microsphere at the point of the AFM probe are contacted with cancer cells and pressed into a measured object for a certain depth d by controlling the z-axis displacement of the atomic force microscope, then the nano particles are separated from the surface of the measured object, when the nano particles on the surface of the micron-sized microsphere are pressed into the measured object and separated from the measured object, magnetic nano particles and the cancer cells generate interaction force, a cantilever of the AFM probe is bent immediately, and laser irradiated on the cantilever of the AFM probe on the probe head of the atomic force microscope generates corresponding deflection and is identified and converted into a force value by a detector on the probe head of the atomic force microscope. And recording the interaction force F (adhesive force) between the magnetic nanoparticles on the carbon microsphere surface of the AFM probe tip and the surface of the cancer cell when the AFM probe tip is separated. Obtaining the contact area S of the magnetic nanoparticles and the cancer cells from the pressing depth d, multiplying the contact area S by the density rho of the magnetic nanoparticles on the surface of the carbon microsphere to obtain the number N of the magnetic nanoparticles participating in the interaction, and dividing the interaction force F (adhesive force) measured by the experiment by the number N of the magnetic nanoparticles to obtain the adhesive force between the unit magnetic nanoparticles and the surface of the object to be measured.
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 (6)
1. An AFM probe for quantitative measurement, characterized in that the probe structure is: a micron-sized small ball is adhered to the tip of the AFM probe, a large number of amorphous concave-convex structures are irregularly distributed on the surface of the micron-sized small ball to form a rough surface, and nano particles are uniformly adhered to the rough surface of the small ball; the roughness of the rough surface is in the nanometer level, and the concave-convex peak-valley value of the rough surface is smaller than the size of the nano particles to be modified.
2. The method for modifying the AFM probe used for quantitative measurement according to claim 1, comprising the following steps:
step 1, pretreatment of micron-sized pellets
(1) The micron-sized globules are pretreated by a chemical etching or mechanical processing method to form a rough surface microstructure which can increase the adsorption effect of the micron-sized globules on the nano-particles; the etching time of the chemical etching is between 0.05 and 2 hours, and the chemical etching adopts a conventional acid washing etching method; the mechanical processing method is a ball milling method by adopting a ball mill, and the ball milling time is between 0.05 and 10 hours; the micron-sized balls are spherical, and the diameter of the micron-sized balls is 0.5-2 mu m;
(2) after the pretreated micron-sized pellets are subjected to ultrasonic treatment, filtering the ultrasonic solution by a filter screen with the diameter equivalent to that of the pellets, and drying the solution in an oven at the drying temperature of between 40 and 120 ℃ for 1 to 24 hours;
step 2, preparing nanoparticle dispersion liquid
Adding the nano particles into a dispersing agent, and carrying out ultrasonic dispersion treatment on the nano particles to obtain nano particle dispersion liquid, wherein the concentration of the nano particle dispersion liquid is 0.1-1 mg/ml; the size of the nano particles is between 10 and 80 nm;
step 3, adsorbing the nano particles by the micron-sized beads
Adding the micron-sized spheres prepared in the step 1 into a nano-particle dispersion liquid, wherein the concentration of the micron-sized spheres is 0.01-0.1mg/ml, performing ultrasonic treatment for 0.2-2h again to obtain a mixed dispersion liquid, and fully contacting nano-particles in the nano-particle dispersion liquid with the micron-sized spheres so as to adsorb the nano-particles on the surfaces of the micron-sized spheres; dripping the mixed dispersion liquid in the center of a silicon chip or a mica sheet, carrying out drying treatment, and placing the silicon chip or the mica sheet in an oven for drying at the drying temperature of 40-200 ℃ for 1-48 h;
step 4, AFM probe modification
At the moment, the nano particles are attached to the surfaces of the micron-sized spheres, the sizes of the micron-sized spheres are clearly visible under an optical microscope, and the micron-sized spheres are attached to the tips of AFM probes through a micromanipulator.
3. The method as claimed in claim 2, wherein the micro-spheres are made of carbon, titanium, silicon dioxide, or polystyrene.
4. The method for modifying the AFM probe for quantitative measurement as claimed in claim 2, wherein the sonication time for the micron-sized bead in step 1 is 0.2-2 h.
5. The method for modifying the AFM probe used for quantitative measurement as claimed in claim 2, wherein the dispersant in step 2 is deionized water or absolute ethanol.
6. The application of the AFM probe used for quantitative measurement as claimed in claim 1, wherein the AFM probe is placed on a probe head of an atomic force microscope, and is used for measuring the interaction force between unit nanoparticles and the surface of a measured object, so that the key adhesion parameters of the nanoparticles and other observed objects can be directly obtained.
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| CN112362909A (en) * | 2020-10-29 | 2021-02-12 | 清华大学 | Pressure sensing method for projecting AFM probe on surface of nanowire in scanning electron microscope |
| CN112989521A (en) * | 2021-04-22 | 2021-06-18 | 四川大学 | Control method based on micro-bending forming size precision multi-objective optimization |
| WO2022227229A1 (en) * | 2021-04-28 | 2022-11-03 | 西安交通大学 | Method for preparing nanoprobe |
| GB2611841A (en) * | 2021-04-28 | 2023-04-19 | Univ Xi An Jiaotong | Method for preparing nanoprobe |
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