CN111551278A - Accurate and rapid temperature measurement system and temperature measurement method for single nanoparticle - Google Patents
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Abstract
The invention discloses a precise and rapid temperature measurement system and a temperature measurement method for a single nanoparticle, wherein the system comprises the following components: the laser emission module comprises a laser generator, a light path adjusting assembly, an optical microscopic amplification objective lens, matching lens oil and a semi-transmitting and semi-reflecting lens, and is used for converting monochromatic incident laser into p-polarized light and emitting the p-polarized light to the sample reaction module at an excited Surface Plasmon Resonance (SPR) angle; the sample reaction module is used for exciting the SPR effect of single nanoparticles; a temperature adjusting module connected with the particle reaction module, a particle intensity detecting module for recording the SPR intensity change of the single nano-particle connected with the sensing chip surface through the biomolecule binding pair at different temperatures to obtain the single nano-particlezA change in direction; by plotting the single nanoparticle temperatures Tsumz 2The temperature of the calibration curve can be accurately and rapidly measured, and the corresponding temperature of the modified biomolecules on the single nanoparticles can be further obtained.
Description
Technical Field
The invention relates to the field of nanoparticle temperature measurement, in particular to a system and a method for accurately and quickly measuring the temperature of a single nanoparticle.
Background
The application of the nano material plays an important role in the progress of nano science and technology, wherein the nano particles are widely applied in the fields of catalysis, information transfer, energy storage and the like. In recent years, due to the continuous development of the field of biochemical analysis, the demand for rapid and sensitive biomolecules has been increasing. Nanoparticles are widely used in medicine and bioengineering as a nano material which is extensively and deeply studied. For example, gold nanoparticles have properties of large surface area ratio, unique optical and electromagnetic properties, easy surface modification and the like, and are widely applied to biological detection and clinical rapid detection. It is well known that changes in temperature can have an effect on the degree of brownian motion of the nanoparticles, which in turn affects their application. Therefore, the method has great significance for accurately and rapidly determining the temperature change of the nanoparticles in the system. However, the current temperature measurement device can only measure the bulk temperature in the bulk phase, and cannot specifically measure the temperature change of the single nanoparticles, which greatly limits the research on the properties and applications of the nanoparticles.
Meanwhile, the method for accurately and rapidly measuring the temperature change of the single nano particle can be expanded to multiple fields, and provides an important basis for development progress of other fields. For example, in the case of studying biomolecular interactions using nanoparticles, most biomolecular interactions are accompanied by endothermic or exothermic processes, and due to the differences between molecules, the interactions between different molecules are different, and thus the temperature changes caused by the interactions between different molecules are also different. The interaction of the biomolecules is researched by utilizing the nanoparticles, and a new research means can be further provided for researching the interaction of the single biomolecules by realizing the measurement of the temperature change of the single nanoparticles.
Disclosure of Invention
In a first aspect, the present invention provides a system for accurate and rapid thermometry of individual nanoparticles, the system comprising:
the laser emission module comprises a laser generator for generating monochromatic incident laser, a light path adjusting component for converting the monochromatic incident laser into p-polarized light, an optical microscopic amplification objective lens for amplifying light path signals and a semi-transmitting and semi-reflecting lens for converting the incident angle of the p-polarized light;
the sample reaction module comprises a sensing chip and a sample cell for placing single nanoparticles, the sample cell, the sensing chip and the optical microscopic amplification objective lens are arranged in the same direction, p-polarized light is incident to the optical microscopic amplification objective lens, the sensing chip and the sample cell at an SPR angle under the action of the semi-transparent semi-reflective lens, the single nanoparticles in the sample cell are excited to generate an SPR effect, and the sensing chip generates a refraction light path and is emitted through the optical microscopic amplification objective lens;
the temperature adjusting module is in control fit with the sample cell to accurately control the temperature of the sample cell;
the particle intensity detection module comprises a reflector for converting the incident angle of a refraction light path emitted by the optical microscopic amplification objective lens and an image sensor for acquiring the image of the light emitted by the reflector;
the temperature of the sample cell was controlled by a temperature adjustment module to record changes in SPR intensity for individual nanoparticles at different temperatures.
The inventors evaluated the effectiveness of the system for monitoring the course of temperature changes; as shown in FIG. 3, the experiment collects the changes of SPR intensity in the deionized water temperature reduction process in the sample pool at different temperatures, obtains the linear relation between the temperature (delta T) and the plasmon resonance angular displacement (delta theta), and the experimental value is-76.4K DEG deg-1，R20.998, and-73.3K deg. calculated from the optical system of SPR angle and water refractive index-1And (5) the consistency is achieved. The results show that the apparatus according to the present invention can accurately and efficiently monitor the temperature change by measuring the change in SPR intensity. Inspired by the experimental results, we can measure the temperature change of single nanoparticles according to the SPR intensity change. Therefore, a set of accurate and rapid temperature measurement system for single nano-particles is built, and meanwhile, the system can be used for measuring the temperature change of the biomolecules modified on the surfaces of the particles.
In some embodiments, the optical path adjusting assembly includes a collimating lens, a polarizer and a condenser lens for converting monochromatic incident laser light into p-polarized light, which is focused on a back focal plane of the optical micro-magnifying objective.
In some embodiments, the optical microscope objective lens and the sensor chip are filled with matching lens oil, and the refractive index of the matching lens oil is 1.51.
In some embodiments, the sample cell is fixed on the upper surface of the sensing chip, and the single nanoparticle surface-modified biomolecule and the sensing chip surface-modified target molecule are combined by adding a solution, so that the single nanoparticle is fixed on the sensing chip surface and brownian motion occurs in the solution. The generated physical optical signal of SPR can be used for sensing the refractive index change of the chip surface, such as the distance between the nano-particles and the sensing chip and the temperature change. The higher the temperature, the more vigorous the corresponding nanoparticle motion.
In some embodiments, the magnification of the optical microscope objective lens is 60, the numerical aperture is 1.49, and the image sensor is a CCD image sensor or a CMOS image sensor.
In some embodiments, the sensing chip is composed of a substrate and a metal layer, the substrate being a BK7 cover glass.
In a second aspect, the invention provides a method for accurately and rapidly measuring temperature of a single nanoparticle, wherein the measuring method adopts the system provided by the invention, and the specific method comprises the following steps: step 1), modifying nanoparticles on the surface of a sensing chip; step 2) acquiring SPR images of the sample at different temperatures to obtain curves of single nanoparticles and SPR image intensity; and testing the SPR image intensity of the sample to be tested, and obtaining the temperature of the single nano-particle through the curve.
In some embodiments, the temperature abscissa, in z, may be obtained by converting the SPR intensity of a single nanoparticle at different temperatures into the distance z of the gold nanoparticle from the surface of the gold plate2Calibration curve on ordinate.
In some embodiments, the step 1) specifically includes the following steps: fixing the sensing chip surface in a sample pool, and bonding the nanoparticles on the sensing chip surface in a single dispersed manner through IgG and Anti-IgG modification
In some embodiments, the method comprises the steps of: the temperature measurement of the modified biomolecules on the single nanoparticles in the step 2) is realized by the following method;
step 21) testing the SPR intensity change of single gold nanoparticles at different temperatures and carrying out detectionThe distance z between the gold nano-particles and the surface of the gold sheet is converted into the distance z, the temperature is taken as the abscissa, and the z is taken as the abscissa2Calibration curve as ordinate; from the relationship of SPR intensity and z: i ═ I0e-z/L(ii) a Wherein, I0The value for SPR intensity when z is 0 is 200, L is the retardation constant, and the value is 100, whereby the z value can be obtained; step 22) mapping the single nanoparticle temperatures T and z2The calibration curve of (1); step 23) according to the calibration curve, we can obtain the change of the nanoparticle in the z direction according to the SPR intensity change of the nanoparticle, and the z is measured2The temperature of the individual nanoparticles was measured by substituting into the curve.
As shown in fig. 4, we obtained that individual nanoparticles bound to the sensor chip surface through IgG and Anti-IgG interactions. The nanoparticles are distributed in a single particle on the surface of the sensor chip through bright field and scanning electron microscope images (fig. 5). The above results demonstrate that individual gold nanoparticles bind to the sensor chip surface through the interaction of IgG and Anti-IgG. Then, the SPR intensity change of single gold nanoparticle at different temperatures is tested and converted into the distance z between the gold nanoparticle and the surface of the gold sheet, and the temperature is taken as the abscissa and the z is taken as the surface of the gold sheet2Calibration curve on ordinate. From the relationship of SPR intensity and z:
I＝I0e-z/L
wherein, I0The value of z is obtained by assuming SPR intensity when z is 0 as 200 and L as a retardation constant as 100.
As shown in fig. 6A, the positional information of the nanoparticles in the z direction was obtained by measuring the SPR intensity change of the nanoparticles at 304K. To show this change more directly, we square the z value, as shown in FIG. 6B. Using this device, we continue to measure SPR intensity values of nanoparticles at other temperatures, obtain the position information of nanoparticles in the z direction (FIG. 6C), and plot the single-nanoparticle temperatures T and z by taking the square of the z value2The calibration curve of (1). As shown in FIG. 7, the nanoparticles increased in position in the z-direction with a slope of 6.02nm as the temperature increased2and/K. From this calibration curve we can follow the nanoparticle SPR intensityVariation, obtaining a variation of the nanoparticle in the z-direction, coupling z2And substituting the curve to obtain the temperature of the gold nanoparticles, the IgG modified on the surface of the gold nanoparticles and the Anti-IgG binding pair modified on the surface of the sensing chip.
The result shows that the device can accurately and rapidly measure the temperature of single nano-particles and modified biomolecules, and provides an important dynamic detection means for researching the influence of the temperature on the Brownian motion of the single nano-particles and the interaction between the biomolecules.
The temperature adjusting module and the plasmon resonance imaging module are combined, the temperature change capability of single nano-particles and surface-modified biomolecules is obtained by directly detecting the change of SPR intensity of the single nano-particles, and a new important research means is provided for researching the local temperature change of the single particles and the interaction of the biomolecules.
The inventor of the invention finds that the SPR intensity is related to the temperature of single nano-particles, and further, the inventor finds that the temperature is used as a horizontal coordinate, and z is used as a horizontal coordinate through SPR intensity change and conversion of the SPR intensity change into the distance z of gold nano-particles from the surface of a gold sheet2Calibration curve as ordinate; and the temperature measurement method is applied to the temperature measurement method of the nano particles, and the temperature measurement result is accurate and reliable.
The method can obtain the local micro temperature change of the single nano particle, and can detect the corresponding temperature of the modified biomolecule binding pair on the single nano particle, specifically the temperature and heat difference generated by the binding, dissociation and intermediate state conformation change of the biomolecule, so as to obtain the related information of biomolecule binding thermodynamics. Through analysis and comparison of different biomolecule combined thermodynamic information on single nano particles, a corresponding biomolecule system thermal energy spectrum can be drawn, and cognition of people in the field of molecular biology is further expanded. Meanwhile, the invention can be expanded to the research of the organism on the temperature adaptation mechanism, such as the analysis of the dynamic temperature change of mitochondria which has important functions on heat production and temperature regulation when the structure and the composition distribution change; can also be expanded to the thermodynamic monitoring of the combination of biomolecules and targets in the medical field, and provides a powerful tool for the early detection and treatment of tumors and cancers.
Drawings
FIG. 1 is a schematic block diagram of a system to which the present invention relates;
FIG. 2 is a flow chart of a method to which the present invention relates;
FIG. 3 is a temperature calibration curve for a device to which the present invention relates;
FIG. 4 is a schematic diagram of the binding of single nanoparticles on a sensor chip according to the present invention:
FIG. 5A is a bright field diagram of a single nanoparticle of the present invention;
FIG. 5B is a scanning electron microscope image of a single nanoparticle according to the present invention;
FIG. 6A is a graph of z at 304K for a single nanoparticle of the present invention;
FIG. 6B shows a single nanoparticle of the present invention at 304K, z2A change in value;
FIG. 6C shows the z-transition of a single nanoparticle of the present invention from 304K to 304.6K2The change of the value is that each section is increased by 0.2K;
FIG. 7 shows the temperatures T and z of individual nanoparticles involved in the present invention2The calibration curve of (1).
Detailed Description
The following examples are given to illustrate the present invention and should not be construed as limiting the scope of the present invention.
Example 1
As shown in fig. 1, the single particle surface plasmon resonance imaging temperature measurement system is set up in the embodiment, and specifically, the system for accurately and rapidly measuring the temperature of a single nanoparticle is provided, and comprises a laser emission module, a sample reaction module, a temperature adjustment module and a particle intensity detection module;
the laser emission module comprises the following four parts: the laser generator is used for generating monochromatic incident laser and is used as an excitation light source of single noble metal nano-particles for generating local plasmon resonance; an optical path adjusting unit 12 for converting monochromatic incident laser light into p-polarized light; the optical microscopic amplification objective lens is used for amplifying optical path signals so as to meet the requirement of carrying out high-magnification imaging analysis on single nanoparticles on the surface of the sensor chip; the semi-transparent semi-reflective lens 15 is used for converting the incident angle of p-polarized light and used for adjusting and optimizing a light path structure to meet the actual requirement that the incident light successfully irradiates an objective lens and a sensing chip; and the matching lens oil 14 is used between the high-power optical micro-magnification objective lens and the sensing chip, and the refractive index of the matching lens oil is close to that of glass so as to meet the requirements of increasing light rays, improving the visual field and obtaining a clear object image.
The light path adjusting component comprises a collimating lens, a polaroid and a condensing lens, and is used for converting monochromatic incident laser into p-polarized light and focusing the p-polarized light on a rear focusing surface of the optical microscopic amplifying objective lens.
The magnification of the optical micro-magnification objective is 60, and the numerical aperture is 1.49;
the sample reaction module comprises a sensing chip 21 and a sample cell 22 for placing single nanoparticles, the sample cell 22, the sensing chip 21 and the optical microscopic amplification objective lens are arranged in the same direction from top to bottom, p-polarized light is incident to the optical microscopic amplification objective lens 13, the sensing chip and the sample cell at an SPR angle under the action of the semi-transparent semi-reflective lens 15, and after the single nanoparticles on the surface of the sensing chip are excited to generate a local plasmon resonance effect, corresponding scattered resonance light is reflected by the optical microscopic amplification objective lens and the reflecting mirror 41 and then is collected by the image sensor 42 to form a corresponding real-time image;
specifically, the sample reaction module is built by using a gold plate and a Polydimethylsiloxane (PDMS) sample pool: the sensing chip is a gold sheet, the gold sheet consists of a substrate and a metal layer, the substrate is a BK7 cover glass, a layer of metal chromium with the thickness of 2nm is plated on the substrate, and a layer of gold with the thickness of 47nm is plated on the substrate; burning the gold sheet by using hydrogen flame before use, then washing the gold sheet by using ethanol and deionized water, and drying the gold sheet by using nitrogen for later use; the gold flakes were modified and soaked overnight with 5mL of 1mM ethanol solution of 11-mercaptoundecanoic acid, in a molar ratio of 4: the mixed solution of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) of 1 activated the carboxyl groups on the surface for 20 minutes. Taking a PDMS (polydimethylsiloxane) pool as a sample pool, washing the sample pool with ethanol and deionized water, drying the sample pool with nitrogen, and placing the sample pool on the surface of a sensing chip for placing a solution.
The temperature adjusting module 3 is matched with the sample cell control to accurately control the temperature of the sample cell; specifically, a heating table 31 is built on an objective table above a micro-amplification objective lens, a sample reaction module is placed in the heating table 31 and above the objective lens, and a heating controller is adjusted to control the temperature of the sample reaction module; in this example, the bulk temperature of the liquid in the sample cell was determined by adding a thermocouple;
the particle intensity detection module 4 comprises a reflector 41 for converting the incident angle of the refraction light path emitted by the optical microscopic amplification objective lens and an image sensor 42 for acquiring the image of the emitted light of the reflector;
controlling the temperature of the sample pool through a temperature regulation module to record the SPR intensity change caused by Brownian motion after the single nano particles are combined on the surface at different temperatures;
specifically, in the embodiment, red laser with a wavelength of 680nm is selected to be introduced into an SPR image to serve as an excitation light source for nanoparticle localized plasmon resonance, a laser generator is turned on, and the intensity is adjusted to 100mA, so that a stronger signal is received on an image sensor (CMOS camera);
selecting a 60X objective lens, and dripping a drop of matching lens oil between the outer end of the optical micro-amplification objective lens and the lower end surface of the sensing chip, wherein the refractive index of the matching lens oil is 1.51; and adjusting the light path adjusting component to enable the incident light to enter the optical microscopic amplifying objective lens and adjust the angle of the incident light to be at the position of exciting the nano particle SPR.
Example 2
The following example of IgG antigen and Anti-IgG antibody binding pairs is used to explore the systems and methods involved in the present invention, as shown in FIG. 2, the specific experimental scheme is:
step 1) calibration of the linear relationship between temperature (Δ T) and plasmon resonance angular displacement (Δ θ): in the single particle SPR imaging heating system set up above, 100 mul of deionized water is added into the PDMS sample cell, a thermocouple is inserted into the sample cell, and the overall temperature of the liquid in the sample cell is measured. 100 mu L of 90 ℃ deionized water is added into the sample cell, the change of SPR intensity in the cooling process of the deionized water in the sample cell is recorded by a CMOS camera, and the shooting speed of the camera is 100 frames per second.
As shown in FIG. 3, the experiment collects the changes of SPR intensity in the deionized water temperature reduction process in the sample pool at different temperatures, obtains the linear relation between the temperature (delta T) and the plasmon resonance angular displacement (delta theta), and the experimental value is-76.4K DEG deg-1，R20.998, and-73.3K deg. calculated from the optical system of SPR angle and water refractive index-1And (5) the consistency is achieved. The results show that the apparatus according to the present invention can accurately and efficiently monitor the temperature change by measuring the change in SPR intensity. Inspired by the experimental results, we can measure the temperature change of single nanoparticles according to the SPR intensity change. Therefore, a set of accurate and rapid temperature measurement system for single nano-particles is built, and meanwhile, the system can be used for measuring the temperature change of the biomolecules modified on the surfaces of the particles.
Step 2) modifying gold nanoparticles on the surface of the sensing chip (taking IgG and Anti-IgG modified gold nanoparticles (150nm) as an example):
step 21) preparation of the sample: accurately weighing 1mg of IgG, and preparing a 100 mu g/mL IgG solution by using 10mL Phosphate Buffer Solution (PBS) as a mother solution; mixing the mother liquor of 2 mu LIgG with 98 mu LPBS, and stirring for 30s on a turbine stirrer for later use; mixing 2 mu of LANti-IgG modified gold nanoparticles (the diameter is 150nm) with 5mL of deionized water, and stirring for 2min on a turbine stirrer for later use; 60mg bovine serum albumin solid powder is accurately weighed, 2mL phosphate buffer solution is accurately measured, and the solution is stirred for 30s by a turbine to prepare bovine serum albumin solution with the mass fraction of 3% for later use.
Step 22) surface modification: adding 100 mu LIgG solution into the sample reaction pool, and standing for 10 min. Subsequently, the sample cell was washed three times with 200 μ L of phosphate Tween buffer solution (PBST, 0.05% Tween); subsequently, 200. mu.L of 3% bovine serum albumin solution was added, and after standing for 10min, the sample cell was washed three times with 200. mu.L of PBST; finally, 100. mu.L of Anti-IgG modified nanoparticle solution was added, and after standing for 10min, the sample cell was washed three times with 200. mu.L of PBST.
As shown in fig. 4, we obtained that individual nanoparticles bound to the sensor chip surface through IgG and Anti-IgG interactions. The nanoparticles are distributed in a single particle on the surface of the sensor chip through bright field and scanning electron microscope images (fig. 5). The above results demonstrate that individual gold nanoparticles bind to the sensor chip surface through the interaction of IgG and Anti-IgG.
Step 3) temperature determination of the biomolecules modified on the individual nanoparticles:
step 31) SPR image intensities of single gold nanoparticles at different temperatures: first, SPR images of nanoparticles at room temperature (298K) were recorded with a camera shooting speed of 100 frames per second for 15 s. And then, regulating and controlling a heating controller, raising the temperature to 4K, keeping for 10min, recording an SPR image of the nanoparticles at the temperature when the temperature of the solution in the sample pool reaches a set temperature of 302K, wherein the shooting speed of a camera is 100 frames per second, and the shooting time is 15 s. And continuously regulating and controlling the heating controller to heat for 2K, keeping for 10min, recording the SPR image of the nano particles at the temperature when the temperature of the solution in the sample pool reaches the set temperature 306K, wherein the shooting speed of the camera is 100 frames per second, and the shooting time is 15 s. And then regulating and controlling the heating controller to heat for 7K, keeping for 10min, recording the SPR image of the nano particles at the temperature when the temperature of the solution in the sample pool reaches the set temperature of 313K, wherein the shooting speed of the camera is 100 frames per second, and the shooting time is 15 s. Finally, regulating and controlling the heating temperature controller to heat up, keeping for 10min, recording the SPR image of the nano-particles at the temperature when the temperature in the sample pool reaches, wherein the shooting speed of a camera is 100 frames per second, and the shooting time is 15 s;
step 32) converting the SPR image intensity change of the single gold nanoparticle at different temperatures into the distance z between the gold nanoparticle and the gold sheet surface, and obtaining the distance z by taking the temperature as a horizontal coordinate and the z as a horizontal coordinate2Calibration curve on ordinate. From the relationship of SPR intensity and z:
I＝I0e-z/L
wherein, I0The value of SPR intensity when z is 0 is 200, L is retardation constant and is 100, and thus the value can be obtainedAnd z is the value.
As shown in fig. 6A, the positional information of the nanoparticles in the z direction was obtained by measuring the SPR intensity change of the nanoparticles at 304K. To show this change more directly, we square the z value, as shown in FIG. 6B. Using this device, we continue to measure SPR intensity values of nanoparticles at other temperatures, obtain the position information of nanoparticles in the z direction (FIG. 6C), and plot the single-nanoparticle temperatures T and z by taking the square of the z value2The calibration curve of (1). As shown in FIG. 7, the nanoparticles increased in position in the z-direction with a slope of 6.02nm as the temperature increased2and/K. From this calibration curve we can derive the change in z direction of the nanoparticle from the change in SPR intensity of the nanoparticle, and apply z to the particle2The temperature of the gold nanoparticles, IgG and Anti-IgG binding pair was measured by substituting the curve.
The result shows that the device can accurately and rapidly measure the temperature of single nanoparticles and modified biomolecules, and provides an important means for researching the influence of the temperature on the Brownian motion of the single nanoparticles and the interaction between the biomolecules.
The above description is only for the specific embodiments of the present invention, and the protection scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be defined by the claims.
Claims (10)
1. An accurate and rapid temperature measurement system for single nanoparticles, comprising:
the laser emission module comprises a laser generator (11) for generating monochromatic incident laser, a light path adjusting component (12) for converting the monochromatic incident laser into p-polarized light, an optical microscopic amplification objective lens (13) for amplifying a light path signal, and a semi-transparent and semi-reflective lens (15) for converting the incident angle of the p-polarized light;
the sample reaction module comprises a sensing chip (21) and a sample cell (22) for placing single nanoparticles, wherein the sample cell, the sensing chip and the optical microscopic amplification objective lens are arranged in the same direction, p-polarized light is incident to the optical microscopic amplification objective lens (13), the sensing chip and the sample cell at an angle of exciting Surface Plasmon Resonance (SPR) under the action of a semi-transparent and semi-reflective lens (15), the single nanoparticles on the surface of the sensing chip are excited to generate a local plasmon resonance effect, and corresponding image signals are collected by an image sensor (42) after scattered light passes through the optical microscopic amplification objective lens and a reflector (41) along a light path;
the temperature adjusting module is in control fit with the sample cell to accurately control the temperature of the sample cell;
the particle intensity detection module comprises a reflector (41) for converting the incidence angle of a scattering light path emitted by the optical microscopic amplification objective lens and an image sensor (42) for acquiring the image of the emitted light of the reflector;
the temperature of the sample cell was controlled by a temperature adjustment module to record changes in SPR intensity for individual nanoparticles at different temperatures.
2. The system for accurate and rapid thermometry of individual nanoparticles according to claim 1, wherein said optical path adjustment assembly (12) comprises a collimating lens (121), a polarizer (122) and a condenser lens (123) for converting monochromatic incident laser light into p-polarized light focused on the back focal plane of an optical micro-magnifying objective.
3. The system for accurate and rapid temperature measurement of single nanoparticle as claimed in claim 1, wherein the matching lens oil (14) is filled between the optical micro-magnifying objective lens (13) and the sensing chip (21), and the refractive index of the matching lens oil is 1.51.
4. The system for accurate and rapid thermometry of individual nanoparticles according to claim 1, wherein the sample cell is fixed on the upper surface of the sensor chip, and the biomolecules surface-modified by the individual nanoparticles and the target molecules surface-modified by the sensor chip are combined by adding the solution, so as to fix the individual nanoparticles on the surface of the sensor chip and generate brownian motion in the solution.
5. The system for accurate and rapid temperature measurement of single nanoparticle as claimed in claim 1, wherein the magnification of the optical microscope objective is 60, the numerical aperture is 1.49, and the image sensor is a CCD image sensor or a CMOS image sensor.
6. The system for accurate and rapid thermometry of individual nanoparticles as claimed in claim 1, wherein the sensing chip is composed of a substrate and a metal layer, the substrate is a BK7 cover glass.
7. A method for accurately and rapidly measuring the temperature of a single nanoparticle, which is characterized in that the system of any one of claims 1-6 is adopted in the measuring method, and the specific method comprises the following steps: step 1), modifying nanoparticles on the surface of a sensing chip; step 2) collecting SPR images of the sample at different temperatures to obtain a brownian motion generation curve and an SPR image intensity curve of a single nanoparticle; and testing the SPR image intensity of the sample to be tested, and obtaining the temperature of the single nano-particle through the curve.
8. The method for accurately and rapidly measuring the temperature of a single nanoparticle as claimed in claim 7, wherein the temperature as abscissa and the z as the abscissa can be obtained by converting the SPR intensity of the single nanoparticle at different temperatures into the distance z between the gold nanoparticle and the surface of the gold sheet2Calibration curve on ordinate.
9. The method for accurately and rapidly measuring the temperature of the single nanoparticle according to claim 7, wherein the step 1) specifically comprises the following steps: fixing the surface of the sensing chip in a sample pool, and bonding the nanoparticles on the surface of the sensing chip in a single and dispersed way through IgG and Anti-IgG biomolecule binding pair modification.
10. The method for accurately and rapidly measuring the temperature of the single nanoparticle as claimed in claim 7, wherein the method comprises the following steps: the temperature measurement of the modified biomolecules on the single nanoparticles in the step 2) is realized by the following method;
step 21) testing the SPR intensity change of the single gold nanoparticle which generates Brownian motion on the surface of the sensing chip through the combination of biomolecules at different temperatures, and converting the SPR intensity change into the vertical distance z of the gold nanoparticle from the surface of the gold sheet, wherein the temperature is used as a horizontal coordinate, and the z is used as the horizontal coordinate2Calibration curve as ordinate; from the relationship of SPR intensity and z: i ═ I0e-z/L(ii) a Wherein, I0The value for SPR intensity when z is 0 is 200, L is the retardation constant, and the value is 100, whereby the z value can be obtained; step 22) mapping the single nanoparticle temperatures T and z2The calibration curve of (1); step 23) according to the calibration curve, we can obtain the change of the nanoparticle in the z direction according to the SPR intensity change of the nanoparticle, and the z is measured2The corresponding temperature of the individual nanoparticles was measured by substituting into the curve.
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