CN109765180B - Medium microsphere auxiliary detection film, preparation method thereof and super-resolution detection method - Google Patents
Medium microsphere auxiliary detection film, preparation method thereof and super-resolution detection method Download PDFInfo
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Abstract
The invention discloses a medium microsphere auxiliary detection film, a preparation method thereof and a super-resolution detection method, wherein the medium microsphere auxiliary detection film comprises a plurality of chambers which are tightly connected, each chamber is filled with a medium microsphere, the chambers and the medium microspheres are packaged in a shell, the uniform periodic arrangement of the microspheres is easy to realize, and the problem of the uniform periodic arrangement of the medium microspheres is solved; the position of the medium microsphere is changed to realize the localized super-resolution detection of the object; the spatial form of the medium microspheres is changed by extending the PDMS film, so that the micro-control of the monitoring performance of the object is realized; the method solves the problem that the medium microspheres have to change the super-resolution in the sphere and at the sphere boundary by a liquid immersion method.
Description
Technical Field
The invention belongs to the technical field of micro-optical detection, and relates to the field of micro-nano manufacturing and silicon dioxide (SiO)2) The manufacture of microspheres and Polydimethylsiloxane (PDMS) films also involves the analysis of physical optics and geometrical optics for detection methods. In particular to a template-induced self-assembly-based medium microsphere auxiliary detection film, a preparation method thereof and a super-resolution detection method.
Background
In the field of nano-photonics, biochips, communication chips, sensor chips, memory chips and the like also put high integration and super-resolution requirements on nano-optics and photonic devices.
The existing super-resolution technology includes super-resolution fluorescence microscopy, structured light illumination microscopy and surface plasma super-resolution. The super-resolution fluorescence microscopy emits fluorescence under the irradiation of a light source with a specific wavelength by means of a fluorescent dye, and reveals the transfer, distribution, positioning and the like of substances in cells. Although this technique can achieve a lateral resolution of 50nm, it requires staining of the sample and its application range is greatly limited. The structured light illumination microscopy technique utilizes a complex structured light field to irradiate a sample to realize tomography, improves the transverse resolution of an optical microscope, but cannot further improve the resolution due to the narrower part of the spatial frequency. The surface plasma super-resolution technology utilizes a standing wave field generated by surface plasma interference to realize super-resolution imaging of an object, but an artificial material with unique dispersion property has huge loss, so that the propagation distance of a light beam in the artificial material is limited, and the imaging magnification and the imaging contrast are limited. Therefore, although these methods can realize super-resolution detection, their applications are limited due to requirements on samples, system construction, operation of algorithms, requirements on materials, and improvement of resolution and contrast.
The super-resolution detection based on the medium microspheres is a super-resolution imaging technology which can break through the optical diffraction limit, and inevitably promotes the development of the subjects of biology, medicine, material science, optics and the like.
The method for realizing the super-resolution of the medium microsphere can be divided into two methods: (1) single microsphere direct interaction imaging; (2) and carrying out immersion type and semi-immersion type medium microsphere super-resolution imaging. The single microsphere direct super-resolution monitoring method cannot realize scanning imaging of fine structures at all positions of an object due to the difficulty in manipulation of the single microsphere. The immersed and semi-immersed medium microsphere super-resolution imaging increases the external refractive index of the microsphere, increases the numerical aperture of the imaging, enhances the possibility of evanescent wave coupling transmission, but reduces the light transmission for imaging by the outer layer liquid, so that the imaging quality is greatly reduced. Therefore, the medium microsphere super-resolution method which is easy to operate and has high imaging quality is a difficult problem which needs to be solved urgently in the super-resolution imaging monitoring direction.
Disclosure of Invention
In order to overcome the defects of difficult manipulation, poor imaging quality and the like of the conventional microsphere super-resolution detection technology, the method for detecting the medium microsphere auxiliary microscope super-resolution based on the template-induced self-assembly technology is provided, and the method can realize the easily-adjusted large-field and far-field super-resolution detection.
In order to achieve the purpose, the medium microsphere auxiliary detection film comprises a plurality of chambers which are tightly connected, wherein each chamber is filled with one medium microsphere, and the chambers and the medium microspheres are packaged in a shell.
Furthermore, the chamber and the housing are made of PDMS.
A preparation method of a medium microsphere auxiliary detection film comprises the following steps:
step 4, embedding the medium microspheres in the cavity of the flexible cavity template by using the uniform dispersion liquid containing the medium microspheres prepared in the step 3 to obtain a flexible film embedded with the medium microspheres;
and 5, packaging the flexible film embedded with the medium microspheres obtained in the step 4 to obtain the medium microsphere auxiliary detection film.
Further, the specific process of step 1 is as follows: firstly removing impurities on the surface of the substrate, and then carrying out ultrasonic oscillation on the substrate in deionized water for 3-5 minutes to ensure that the surface of the substrate has hydrophilicity.
Further, the specific process of step 2 is as follows: pouring the liquid flexible material on the substrate processed in the step (1), and controlling the film thickness of the flexible material by adjusting the rotating speed of the spin coater; then vacuumizing, and eliminating air bubbles generated in the flexible material during stirring and pouring to obtain a flexible material film; then pressing the grating on the flexible material film, and curing the flexible material film; and then stripping the grating from the PDMS film to obtain the flexible chamber template with the periodic chamber.
Further, the refractive index of the flexible material in the step 2 is greater than 1.4.
Further, in the step 3, the refractive index n of the dielectric microsphere1And refractive index n of the flexible material2Satisfies the following conditions: 1<n1/n2<1.2。
Further, the specific process of step 4 is as follows: dropping deionized water dispersion liquid containing medium microspheres on a flexible chamber template; then vacuumizing is carried out, so that the dispersion liquid is filled in the cavity of the flexible cavity template; the substrate is inclined, and the medium microspheres are filled into the chamber under the action of self gravity along with the evaporation of the moisture.
A microscope super-resolution detection method based on a medium microsphere auxiliary detection film comprises the following steps:
3, stretching the medium microsphere auxiliary detection film, changing the shape of medium microspheres in the medium microsphere auxiliary detection film, and realizing super-resolution imaging of the object to be detected in different degrees;
and 4, repeating the steps 1 to 3 until clear super-resolution imaging patterns are observed by the medium microspheres in the medium microsphere auxiliary detection film.
Further, in step 1, the distance between the medium microsphere auxiliary detection film and the object to be detected meets the requirement of evanescent wave coupling transmission.
Compared with the prior art, the invention has at least the following beneficial technical effects:
a layer of medium microspheres is uniformly filled in a medium microsphere auxiliary detection film, and a medium microsphere auxiliary microscope can be used for carrying out super-resolution detection.
A preparation method of a medium microsphere auxiliary detection film utilizes a flexible film to fix medium microspheres, is easy to realize uniform periodic arrangement of the microspheres, overcomes the difficulty and stability of increasing farther focal distance in other super-resolution methods, and solves the problem of uniform periodic arrangement of the medium microspheres; the invention only adopts the template to induce self-assembly, glue homogenizing, separating and thermoplastic curing of the flexible material to prepare the film which can meet the requirements. Compared with the super-resolution fluorescence microscopy, the method does not need to dye the object to be detected and a complex system positioning algorithm; compared with the structured light illumination microscopy, the method does not need to manufacture a complex structured light field, and can further improve the resolution; compared with the surface plasma super-resolution technology, the method does not need artificial materials with unique dispersion properties, has insignificant loss of imaging light waves and is not limited by imaging contrast.
A medium microsphere auxiliary microscope super-resolution detection method based on template-induced self-assembly technology enables a microscope imaging area to project more medium microspheres and more light rays to enter an objective lens, and super-resolution detection of a large field of view is achieved; meanwhile, the medium microspheres are periodically embedded in the chamber, so that the positions of the medium microspheres and the microscope objective lens can be conveniently adjusted, and the super-resolution detection of other parts of the object is realized. The shape of the medium microspheres is changed by extending the flexible film wrapped with the medium microspheres, so that micro-control on the super-resolution imaging performance of the object to be detected can be realized. The problem that the single medium microsphere is difficult to manipulate when used for super resolution is solved.
Further, using dielectric microspheres (n)1＝1.46，n1Is SiO2Refractive index of (d) is uniformly embedded in the flexible material, since the refractive indices of both materials satisfy: 1<n1/n2<1.2, so at the microsphere interface, evanescent waves are more easily coupled into the medium microspheres for propagation. The super-resolution capability at the microsphere interface is increased. The method solves the problem that the medium microsphere must change the super-resolution in the sphere and at the sphere boundary by a liquid immersion method.
Drawings
FIG. 1 is a schematic view of a process for substrate processing according to the present invention;
FIG. 2 is a schematic view of the casting and imprinting process of the present invention;
FIG. 3 is a schematic diagram of the template induction based microsphere autonomous assembly process of the present invention;
FIG. 4 is a schematic diagram of a process for packaging a dielectric microsphere-embedded film according to the present invention;
FIG. 5 is a schematic cross-sectional view of a dielectric microsphere-assisted detection film;
FIG. 6 is a schematic diagram of the present invention for super-resolution detection of an analyte;
FIG. 7 is a schematic illustration of the micro-deformation of the media microspheres of the present invention;
FIG. 8 is a clear super-resolution imaging effect diagram of the dielectric microsphere of the present invention.
In the drawings: 1. the detection device comprises a substrate, 2, a heating platform, 3, a beaker, 4, a first cleaning solution, 5, deionized water, 6, an ultrasonic oscillator, 7, an ultrasonic oscillation generator, 8, a second cleaning solution, 9, a flexible material film, 10, a grating bottom, 11, a grating grid line, 12, a grating transparent part, 13, a cavity, 14, a medium microsphere, 15, an object to be detected, 16, an eyepiece, 17, an objective lens, 18, a flexible cavity template, 19 and a medium microsphere auxiliary detection film.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention. Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified. In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Referring to fig. 4 and 5, a dielectric microsphere auxiliary detection film comprises a plurality of closely connected chambers 13, wherein each chamber 13 is provided with a dielectric microsphere 14, and the chambers 13 and the dielectric microspheres 14 are encapsulated in a shell 20. Wherein, the chamber 13 and the housing 20 are both made of PDMS, and the chamber 13 is a regular hexahedron with a side length of 30 μm.
A preparation method of a medium microsphere auxiliary detection film comprises the following steps:
step 4, embedding medium microspheres 14 in a cavity 13 of a flexible cavity template by utilizing a template induced self-assembly technology to obtain a flexible film embedded with the medium microspheres 14;
and 5, packaging the flexible film embedded with the medium microspheres 14 to obtain a medium microsphere auxiliary detection film, wherein the medium microsphere auxiliary detection film 19 can realize the operability of super-resolution monitoring and the large-field-of-view high-frequency information transmission of the medium microspheres.
As shown in fig. 1, in step 1, the substrate 1 is processed as follows:
a square quartz having a length of 20mm and having good flatness and smoothness was selected as the substrate 1. Placing the substrate in a beaker 3 with a first cleaning solution 4 prepared from 98% concentrated sulfuric acid and hydrogen peroxide in a mass ratio of 3:1, heating the substrate to 80 ℃ by using a heating platform 2, and keeping the temperature of 80 ℃ for one hour to remove impurities on the surface of the substrate 1; taking out the substrate 1, repeatedly washing with deionized water 5, and ultrasonically oscillating in the deionized water 5 for 3-5 minutes to remove the acidic cleaning solution 4 on the surface of the substrate 1; the method comprises the steps of placing a substrate 1 into a second cleaning solution 8, preparing the second cleaning solution 8 from ammonia water, hydrogen peroxide and deionized water in a mass ratio of 1:1:5, carrying out ultrasonic oscillation for one hour by using an ultrasonic oscillator 6 and an ultrasonic oscillation generator 7, taking out the substrate 1, repeatedly washing the substrate 1 with the deionized water for 2-3 times, and carrying out ultrasonic oscillation for 3-5 minutes in the deionized water, so that the surface of the substrate 1 has hydrophilicity.
The step 1 is to process the substrate 1, so that the flexible material can be uniformly dispersed into a micron-scale film before curing and forming, which is the imaging distance requirement necessary for the super-resolution of the dielectric microsphere.
As shown in fig. 2, in step 2, the casting and stamping process is as follows:
the flexible material is desirably a high refractive index, high transparency, curable low surface energy material, preferably dimethyl siloxane, as shown in fig. 2. And (3) mixing and stirring the Dow Corning 184A solution and the Dow Corning 184B solution uniformly according to the volume ratio of 10:1, pouring the mixture on the substrate 1 treated in the step (1), and controlling the film thickness of the PDMS film by adjusting the rotating speed of a spin coater. And vacuumizing, and eliminating tiny air bubbles generated in the PDMS during stirring and pouring to improve the uniformity and light transmittance of the PDMS so as to obtain the PDMS film. Pressing a grating on the PDMS film, wherein the grating comprises a grating bottom 10, a grating grid line 11 and a grating transparent part 12, when the grating is pressed on the PDMS film, the grating bottom is positioned above, and the grating grid line 11 is positioned below, namely, the grating is inverted, and the grating with the light-tight part 11 of about 30 mu m is pressed on the PDMS film. The PDMS film and the grating were then cured in an oven at 80 ℃ for two hours. Then the grating is peeled off from the PDMS film, and the flexible cavity template 18 with the periodic cavities 13 is obtained, wherein the cavities 13 are tightly connected on the same plane, and the side length of the cross section of each cavity 13 is a square of 30 μm. Finally, a layer of octafluorocyclobutane is sprayed on the periodic chamber 13. The chamber 13 may enable dispersion of the media microspheres 14 into the chamber 13 for super-resolution imaging. Spraying C on the chamber 134F8In order to provide the chamber 13 with a superhydrophobic surface.
The medium microspheres in the step 3 must be uniformly dispersed in the solution;
the medium microspheres are solid hard microspheres with high refractive index, high light transmittance, low price and easy obtainment, the diameter of the microspheres is generally not more than 30 mu m, and the silicon dioxide microspheres are selected. The dispersant for the dielectric microspheres generally requires a high purity, less charged ionic solvent, where deionized water is the preferred solvent. And (3) placing the silicon dioxide microspheres with the diameter not more than 30 mu m into deionized water, and carrying out ultrasonic oscillation to uniformly disperse the silicon dioxide microspheres in the deionized water to obtain a uniform dispersion liquid containing the medium microspheres.
As shown in fig. 3, the template in step 4 induces self-assembly, so that the medium microspheres are uniformly dispersed in the chamber 13;
the adhesion between the silica microspheres 14 and the PDMS9 and the surface energy difference of the micro-components are very important for the accuracy of template-induced self-assembly. Most structural substrates have uniform surface chemical energy, and often control of surface energy differences also involves complex surface treatment processes.
The silica deionized water dispersion was sucked up with a pipette and dropped onto the PDMS chamber template. Due to the large surface tension of the hydrophobic microstructure surface and the silica dispersion, the silica dispersion is in a hydrophobic state on the PDMS chamber template. By the action of vacuum pumping, the dispersion liquid is filled in the cavity 13 of the PDMS cavity template, so that the PDMS cavity template has a similar effect of super-hydrophilicity in the cavity 13, and a new flexible cavity template with a hydrophilic-hydrophobic phase is obtained. As the dispersion flows over the "hydrophilic-hydrophobic" interphase structure, the substrate template will, through the effect of template-induced self-assembly, cause the silica microspheres 14 to fill the periodic cavities 13 of the substrate template with the silica deionized water dispersion.
Slightly tilting the substrate 1, due to the larger diameter of the silica microspheres 14 plus the higher density of the microspheres themselves, a close packing of silica microspheres 14 will appear in the PDMS chamber 13 as the water evaporates and under the action of the gravity of the microspheres themselves.
In the step 5, the medium microspheres 14 need to be encapsulated, so that the medium microspheres 14 are fixed in the cavity 13 and have a solid immersion effect; the thickness of the flexible film embedded with the medium microspheres is 30-40 μm.
Referring to fig. 4, a layer of PDMS is poured outside the flexible film chamber embedded with the dielectric microspheres 14, and then vacuum is applied to wrap the dielectric microspheres 14 in the PDMS. The PDMS film thickness is controlled by adjusting the rotating speed of the spin coater, and then the PDMS film is placed into an oven with the temperature of 80 ℃ for curing for two hours. And finally, stripping the cured PDMS from the substrate to form a film with the silicon dioxide microspheres which are uniformly and periodically arranged.
The whole thickness of the PDMS film cannot be too thick, so that the micro objective lens and the sample to be measured are enough adjustable to be completely stripped from the substrate 1.
The super-resolution microscopic detection method based on the medium microsphere auxiliary detection film comprises the following steps:
and 4, repeating the steps 1 to 3 until clear super-resolution imaging patterns are observed through the medium microspheres 14 in the medium microsphere auxiliary detection film.
As shown in fig. 6, in step 1, the flexible PDMS film needs to be closely attached to the surface of the object to be measured, and observed by using an optical microscope. The arrows in fig. 6 are light rays.
The distance between the upper plane of the mechanical table and the lower plane of the mechanical table is adjusted to press the blue-ray disc 15 of the object to be measured on the flexible film, so that the sample is contacted with the flexible film, the distance between the film and the object to be measured meets the requirement of evanescent wave coupling transmission, is generally less than 10 mu m, and the film is placed in a vacuum environment to ensure that the flexible film is fully and tightly pasted on the surface of the object to be measured.
And adjusting the objective lens left and right to enable the objective lens to be vertically opposite to the medium microspheres in the film. Thus, the vertical light entering the microsphere can improve evanescent wave coupling propagation. The medium microsphere focuses the light wave at a farther focus f-nr/2 (n-n)s). Wherein n and nsRespectively, the refractive indexes outside the medium microsphere and the microsphere, and r is the radius of the medium microsphere. The objective lens is adjusted up and down to make the appearance in the microscope most clearWhen the aperture angle 2 α between the objective lens and the sample is the largest, the numerical aperture (NA ═ n · sin α, where n is the refractive index between the objective lens and the object to be measured) of the objective lens is the largest, and the minimum distance between two object points resolvable in the microscope reaches the minimum value (σ ═ 0.61 λ/NA, λ is the wavelength of the light source), it is possible to theoretically resolve the object on the object<Ultra-fine structure of 200 nm.
In the step 2, the film is moved left and right, the position under the medium microspheres can be adjusted, and the super-resolution imaging of different fine structures of the object to be detected under the medium microspheres is realized, so that the super-resolution imaging information of different parts of the object can be monitored;
this step is typically accomplished using tweezers or a micro-motion stage to move the PDMS film side to side or up and down.
As shown in fig. 7, in step 3, the film is stretched to change the shape of the medium microsphere, so as to change the super-resolution imaging effect of the medium microsphere to adapt to the distance between the film and the object to be measured, thereby achieving different super-resolution effects of the microsphere. After stretching the film, the dielectric microspheres 14 are subjected to a tensile force and will deform. The arrow direction in the figure is the direction of the pulling force. The step is realized by stretching the PDMS film embedded with the medium microspheres left and right by using a micro-clamp platform capable of exerting force bidirectionally.
And (5) repeating the steps 1-3 to find clear super-resolution imaging of the object to be detected, as shown in figure 8. The grid distance D of the object to be measured on the blue-ray disc 15 is less than 200 nm.
The invention is easy to realize the uniform periodic arrangement of the microspheres and solves the problem of the uniform periodic arrangement of the medium microspheres; the position of the medium microsphere is adjusted to realize the localized super-resolution detection of the object; the spatial form of the medium microspheres is changed by extending the PDMS film, so that the micro-control of the monitoring performance of the object is realized; the method has simple process, and solves the problem that the medium microspheres have to change the super-resolution in the sphere and at the sphere boundary by a liquid immersion method.
Finally, it should be noted that: the above description is only an example of the present invention and is not intended to limit the present invention. For the present invention, the controllable medium microsphere super-resolution technology is not limited to the above. Although the present invention has been described in detail with reference to the foregoing examples, it will be apparent to those skilled in the art that various changes in the form and details of the embodiments described herein may be made, and equivalents may be substituted for elements thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A preparation method of a medium microsphere auxiliary detection film is characterized by comprising the following steps:
step 1, processing a substrate (1) to enable the substrate (1) to have a super-hydrophilic surface;
step 2, manufacturing a flexible material film (9) on a substrate (1) with a super-hydrophilic surface, and manufacturing periodically arranged chambers (13) on the flexible material film (9) by using a pouring method and an imprinting method to obtain a flexible chamber template (18);
step 3, placing the medium microspheres (14) in a dispersing agent, and uniformly stirring to obtain a uniform dispersion liquid containing the medium microspheres (14);
step 4, embedding the medium microspheres (14) into the cavity (13) of the flexible cavity template (18) by using the uniform dispersion liquid containing the medium microspheres (14) prepared in the step 3 to obtain a flexible film embedded with the medium microspheres (14);
step 5, packaging the flexible film embedded with the medium microspheres (14) obtained in the step 4 to obtain a medium microsphere auxiliary detection film (19);
the medium microsphere auxiliary detection film prepared in the steps 1 to 5 comprises a plurality of chambers (13) which are tightly connected, each chamber (13) is filled with one medium microsphere (14), and the chambers (13) and the medium microspheres (14) are packaged in a shell (20).
2. The preparation method of the dielectric microsphere auxiliary detection film as claimed in claim 1, wherein the specific process of step 1 is as follows: firstly removing impurities on the surface of the substrate (1), and then ultrasonically oscillating the substrate (1) in deionized water for 3-5 minutes to enable the surface of the substrate (1) to have hydrophilicity.
3. The method for preparing the dielectric microsphere auxiliary detection film according to claim 1, wherein the specific process of the step 2 is as follows: pouring the liquid flexible material onto the substrate (1) treated in the step (1), and controlling the film thickness of the flexible material through the rotating speed of a glue spinning machine; then vacuumizing, eliminating air bubbles in the flexible material to obtain a flexible material film (9); then pressing the grating on the flexible material film (9), and curing the flexible material film (9); and then stripping the grating from the PDMS film to obtain the flexible chamber template (18) with the periodic chamber (13).
4. The method for preparing a dielectric microsphere auxiliary detection film as claimed in claim 1, wherein the refractive index of the flexible material in the step 2 is greater than 1.4.
5. The method for preparing a dielectric microsphere auxiliary detection film as claimed in claim 1, wherein in the step 3, the refractive index n of the dielectric microspheres (14)1And refractive index n of the flexible material2Satisfies the following conditions: 1<n1/n2<1.2。
6. The method for preparing the dielectric microsphere auxiliary detection film according to claim 1, wherein the specific process of the step 4 is as follows: dropping a dispersion of deionized water containing media microspheres (14) onto a flexible chamber template (18); then vacuumizing is carried out, so that the deionized water dispersion liquid containing the medium microspheres (14) is filled in the cavity (13) of the flexible cavity template (18); the substrate (1) is inclined, and the medium microspheres (14) are filled into the chamber (13) under the action of self gravity along with the evaporation of the moisture.
7. The microscope super-resolution detection method for the medium microsphere auxiliary detection film prepared by the preparation method according to claim 1 is characterized by comprising the following steps of:
step 1, attaching a medium microsphere auxiliary detection film (19) to the surface of an object to be detected (15), adjusting the distance between the medium microsphere auxiliary detection film (19) and the object to be detected (15), and observing by using an optical microscope to obtain super-resolution imaging of the object to be detected (15);
step 2, moving the medium microsphere auxiliary detection film (19), and monitoring super-resolution imaging information of different parts of the object (15) to be detected;
step 3, stretching the medium microsphere auxiliary detection film (19), changing the shape of the medium microspheres (14) in the medium microsphere auxiliary detection film (19), and realizing super-resolution imaging of the object to be detected (15) in different degrees;
and 4, repeating the steps 1 to 3 until clear super-resolution imaging patterns are observed through the medium microspheres (14) in the medium microsphere auxiliary detection film (19).
8. The method for microscope super-resolution detection based on the dielectric microsphere auxiliary detection film as claimed in claim 7, wherein in step 1, the distance between the dielectric microsphere auxiliary detection film (19) and the object (15) to be detected meets the requirement of evanescent wave coupling transmission.
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