Super-resolution microscope objective, manufacturing method and super-resolution microscopic device

Abstract

A super-resolution microscope objective, a manufacturing method and a super-resolution microscopic device. A plano-convex lens (16) is manufactured on a glass sheet in a controllable preparation mode; then by means of precise micro-control and thermal / light curing, a microsphere (22) is controllably positioned and fixed on the upper surface of the central position of the plano-convex lens (16); and the glass sheet is integrated with a sleeve and mounted on an objective lens (27) to form a microsphere (22) assisted super-resolution microscope objective. According to the super-resolution microscope objective provided, water or oil is added above the glass sheet, and thus would not contact with a sample and has no potential pollution or impact; compared with other methods of directly placing the microsphere (22) on a plane or a flat plate, placing the microsphere (22) on the plano-convex lens (16) can effectively avoid the problem that during microsphere (22) assisted imaging, the other positions of the objective lens contact with the sample earlier, so that the microsphere (22) cannot achieve imaging; and the plano-convex lens (16) can be adjusted according to requirements, and the plano-convex lens (16) has certain enhancement effect on the imaging effect of the microsphere (22). In addition, the manufacturing method is simple and easy to control and can be easily operated and implemented, and the performance of a conventional optical microscope can be improved at low cost.

Description

Super-resolution microscope objective lens, manufacturing method and super-resolution microscope device Technical Field

[0001] The present application relates to the technical field of optical microscope objective lenses, and in particular to a super-resolution microscope objective lens, a manufacturing method, and a super-resolution microscope device. Background Art

[0002] Due to the Abbe diffraction limit, the resolution of traditional optical microscopes is approximately half the wavelength. This makes it impossible to image objects with a size below the diffraction limit. With the continuous development of science and technology, there are many methods / techniques that can break the diffraction limit. Among them, microsphere-assisted super-resolution imaging is a relatively simple and easy method. It achieves super-resolution imaging by combining a conventional optical microscope with a microsphere lens. It has the characteristics and advantages of being label-free, low-cost, fast imaging speed, and easy to integrate and assemble. So far, this method has been widely used in biology, chemistry, materials, medicine and other fields, and has achieved certain results, becoming one of the important driving forces for the development of super-resolution imaging related fields.

[0003] In the field of microsphere-assisted super-resolution imaging technology, the use of high numerical aperture objectives such as water-immersion or oil-immersion objectives in combination with microspheres can enable microscopic imaging systems to achieve greater resolution and magnification capabilities. However, the use of water-immersion or oil-immersion objectives, especially for systems in reflective imaging mode, requires the addition of water or oil to the sample during use, which is bound to cause potential contamination and impact on the sample. In addition, due to the small size of the microspheres, their imaging working distance is within the range of hundreds of nanometers. The existing microsphere-objective integration method is to fix the microspheres directly on the glass front lens of the objective lens. In actual imaging, due to factors such as the rough surface of the sample and the difficulty in positioning the sample surface or the glass front lens surface of the objective lens into an ideal horizontal plane, other parts of the objective lens will contact the sample first, causing the microspheres to be unable to reach their effective imaging distance and thus unable to assist in super-resolution imaging.

[0004] Summary of the Invention

[0005] In view of this, it is necessary to provide a super-resolution microscope objective lens, a manufacturing method and a super-resolution microscope device that can realize real-time and rapid super-resolution imaging of sub-diffraction limit scale objects to address the defect that existing objective lenses cannot assist in achieving super-resolution imaging.

[0006] To solve the above problems, this application adopts the following technical solutions:

[0007] One of the purposes of this application is to provide a method for manufacturing a super-resolution microscope objective lens, comprising the following steps:

[0008] Prepare plano-convex lenses on glass slides;

[0009] transferring a transparent polymer droplet to the center of the plano-convex lens;

[0010] fixing microspheres to the transparent polymer droplets;

[0011] The glass slide with the microspheres fixed thereon is fixed to an objective lens to obtain the super-resolution microscope objective lens.

[0012] In some embodiments, the step of preparing a plano-convex lens on a glass sheet specifically includes the following steps:

[0013] Providing a glass sheet, and placing a nylon sheet with a central hole on the glass sheet;

[0014] Transferring a transparent polymer droplet into the central hole of the nylon sheet and naturally flattening it to form a plano-convex lens;

[0015] The glass sheet is thermally cured to form a solid plano-convex lens.

[0016] In some embodiments, the step of transferring the transparent polymer droplet into the central hole of the nylon sheet and naturally tiling the droplet to form a plano-convex lens specifically includes the following steps:

[0017] A transparent polymer is adsorbed by an adsorbent with a fine tip and transferred into the central hole of the nylon sheet, where it is naturally flattened to form a plano-convex lens. The adsorbent includes an acupuncture needle, an AFM probe, or a semi-tapered optical fiber, and the transparent polymer includes polydimethylsiloxane, polymethyl methacrylate, or UV-curable glue.

[0018] In some embodiments, the step of thermally curing the glass sheet to form a solid plano-convex lens specifically includes the following steps:

[0019] The glass sheet is placed in a drying machine and thermally cured to form a solid plano-convex lens. The curing temperature is 40° C. to 100° C., and the curing time is 20 minutes to 3 hours.

[0020] In some embodiments, the thickness and curvature radius of the plano-convex lens are controlled by controlling the amount of the transparent polymer and the size of the central hole of the nylon sheet.

[0021] In some embodiments, the step of transferring the transparent polymer droplet to the center of the plano-convex lens specifically includes the following steps:

[0022] Obtain two sections of semi-tapered optical fiber;

[0023] A small amount of transparent polymer droplets is added to a section of a semi-tapered optical fiber and allowed to slide from the tapered transition zone to the tapered waist under the action of gravity. The transparent polymer droplets attached to the tapered waist area of ​​the tapered optical fiber form tiny spindle-shaped droplets under the action of solid-liquid interfacial tension.

[0024] fixing the semi-tapered optical fiber with the spindle-shaped droplet attached to a three-dimensional translation stage;

[0025] Transferring one or more spindle-shaped droplets to the tip of a semi-tapered optical fiber fixed on another section of the three-dimensional translation stage by adsorption transfer to form a single tiny droplet;

[0026] The tiny droplet is transferred to the center of the plano-convex lens by the three-dimensional displacement stage.

[0027] In some embodiments, the step of obtaining two sections of semi-tapered optical fibers specifically includes the following steps:

[0028] A single-mode optical fiber with a coating stripped from the middle is fixed at one end to an optical rod and at the other end to a three-dimensional translation stage. The stripped bare optical fiber is heated, and the translation stage knob is rotated to stretch the optical fiber to form a tapered optical fiber. The optical fiber is further stretched using the translation stage until the optical fiber breaks, resulting in two semi-tapered optical fibers. The heating can be performed using a butane torch, an arc lighter, an alcohol lamp, or a carbon dioxide laser.

[0029] In some embodiments, the step of fixing the microspheres to the transparent polymer droplets specifically includes the following steps:

[0030] A suspension containing microspheres is dripped onto a glass slide, and then dried and heated to evaporate the liquid to leave the microspheres. The microspheres include hemispheres, spherical segments, spherical caps, and deformed microspheres. The microspheres are made of a high refractive index material, such as barium titanate glass, chalcogenide glass, or tellurite glass.

[0031] Obtaining a section of optical fiber, stripping the coating layer of the section of optical fiber at the end portion thereof, fixing the section of optical fiber to the glass slide so that the bare optical fiber stripped of the coating layer extends beyond the outer boundary of the glass slide, and then fixing the glass slide to the three-dimensional translation stage;

[0032] Manipulating the three-dimensional translation stage to make the tip of the bare optical fiber contact the single microsphere, and the microsphere will be adsorbed on the tip of the bare optical fiber;

[0033] placing a glass slide with the plano-convex lens under the observation field of view, manipulating the three-dimensional translation stage to position and transfer the microsphere onto the transparent polymer droplet;

[0034] The glass sheet with the plano-convex lens and the microspheres is solidified so that the microspheres are fixed to the transparent polymer droplets.

[0035] In some embodiments, in the step of manipulating the three-dimensional translation stage to make the tip of the bare optical fiber contact the single microsphere and the microsphere is adsorbed onto the tip of the bare optical fiber, the size of the microsphere is several microns to tens of microns.

[0036] In some embodiments, the step of curing the glass sheet with the plano-convex lens and the microspheres so that the microspheres are fixed to the transparent polymer droplets specifically includes: placing the glass sheet with the plano-convex lens and the microspheres under ultraviolet light for curing so that the microspheres are fixed to the transparent polymer droplets, wherein the illumination time is 10-60 minutes, and the irradiation power range of the ultraviolet lamp is 30W-100W.

[0037] In some embodiments, the step of fixing the glass sheet with the microspheres fixed thereon to the objective lens to obtain the super-resolution microscope objective lens specifically includes the following steps:

[0038] A sleeve is provided, which includes a fixed part and a rotating part. The fixed part is nested and fixed on the objective lens, and the rotating part is sleeved on the fixed part and can rotate around the fixed part. The rotating part includes a hollow cylinder and the ring fixedly connected to the hollow cylinder. The inner wall of the hollow cylinder is provided with a thread. By rotating the thread, the rotating part can be adjusted to move up and down along the fixed part.

[0039] The glass sheet with the microspheres fixed thereon is fixed to the circular ring and heated and cured, and then the fixed part of the sleeve is fixed to the objective lens. Finally, the rotating part of the sleeve containing the circular ring is fixed to the fixed part of the sleeve, that is, the sleeve including the glass sheet with the microspheres is fixed to the objective lens to obtain the super-resolution microscope objective lens.

[0040] In some embodiments, the rotating part further includes a support column, which is fixedly connected to the ring and the hollow cylinder.

[0041] In some embodiments, the step of fixing the glass sheet with the microspheres fixed thereon to the ring, heating and curing the ring, and fixing the glass sheet with the microspheres to the objective lens to obtain the super-resolution microscope objective lens specifically includes the following steps:

[0042] A small amount of colloidal material is sucked up with a pipette and applied on the ring. A glass sheet with the microspheres fixed thereon is placed on the ring and placed in a dryer for heating and curing to fix the glass sheet to the sleeve to obtain the super-resolution microscope objective lens.

[0043] In some embodiments, the objective lens includes a water immersion objective lens or an oil immersion objective lens.

[0044] The second purpose of this application is to provide a super-resolution microscope objective lens, which is prepared by the preparation method described above.

[0045] The third object of this application is to provide a super-resolution microscope device, including the super-resolution microscope objective lens.

[0046] This application adopts the above technical solution, and its beneficial effects are as follows:

[0047] The super-resolution microscope objective lens, manufacturing method, preparation method and super-resolution microscope device provided by the present application are to make a plano-convex lens on a glass sheet in a controllable manner, and then the microspheres are controllably positioned and fixed on the upper surface of the center position of the plano-convex lens by precise micro-manipulation and heat / light curing, and integrated with the sleeve and assembled to the objective lens to form a microsphere-assisted super-resolution microscope objective lens. The super-resolution microscope objective lens provided by the present application, adding water or oil on top of the glass sheet, will not contact the sample, and there is no potential pollution or impact. Compared with other methods of directly placing microspheres on a plane or a flat plate, the present application places the microspheres on the plano-convex lens, which can effectively avoid the problem that other places of the objective lens first contact the sample when the microspheres assist imaging, causing the microspheres to be unable to achieve imaging. At the same time, the plano-convex lens can be adjusted as needed, and the plano-convex lens has a certain enhancing effect on the imaging effect of the microspheres. In addition, the above-mentioned manufacturing method is simple and easy to control, easy to operate and implement, and can improve the performance of conventional optical microscopes at a low cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] FIG1 is a flow chart of the steps of a method for manufacturing a super-resolution microscope objective lens provided by one embodiment of the present invention;

[0049] FIG2 is a schematic diagram of a process for preparing a plano-convex lens on a glass sheet according to an embodiment of the present application;

[0050] FIG3 is a schematic diagram of a process for obtaining two sections of semi-tapered optical fibers according to an embodiment of the present application;

[0051] FIG4 is a schematic diagram of a process of transferring a UV-curable adhesive to a plano-convex lens according to an embodiment of the present application;

[0052] FIG5 is a schematic diagram of a process for fixing microspheres to the transparent polymer droplets according to an embodiment of the present application;

[0053] FIG6 is a top view of a plano-convex lens after placement of microspheres according to an embodiment of the present application;

[0054] FIG7 is a side view (left) of a polydimethylsiloxane plano-convex lens and a side view (right) after microspheres are placed therein, provided in one embodiment of the present application.

[0055] FIG8 is a schematic structural diagram of a sleeve provided in an embodiment of the present application.

[0056] FIG9 is a product diagram of a sleeve provided in one embodiment of the present application.

[0057] FIG10 is a diagram showing the preparation process of fixing the glass sheet with the microspheres fixed thereon to the objective lens to obtain the super-resolution microscope objective lens provided in one embodiment of the present application. DETAILED DESCRIPTION

[0058] The following describes in detail embodiments of the present application, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present application, and should not be construed as limiting the present application.

[0059] In the description of this application, it should be understood that the terms "upper", "lower", "horizontal", "inside", "outside", etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing this application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on this application.

[0060] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of the technical features being referred to. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of the features. Throughout the description of this application, "plurality" means two or more, unless otherwise specifically defined.

[0061] In order to make the purpose, technical solutions and advantages of this application more clear, this application is further described in detail below with reference to the accompanying drawings and embodiments.

[0062] Please refer to FIG1 , which is a flowchart of a method for manufacturing a super-resolution microscope objective lens provided in an embodiment of the present application, including steps S110 to S140 . The implementation of each step is described in detail below.

[0063] Step S110: preparing a plano-convex lens on a glass sheet.

[0064] In this embodiment, a schematic flow chart of preparing a plano-convex lens on a glass sheet is provided, which specifically includes the following steps S111 to S113 , and the implementation method of each step is described in detail below.

[0065] Step S111: providing a glass sheet, and placing a nylon sheet with a central opening on the glass sheet.

[0066] In this embodiment, the glass sheet provided in this embodiment can be replaced by other processable hard transparent materials, such as organic glass sheet.

[0067] Step S112: transferring the transparent polymer droplet into the central hole of the nylon sheet and naturally flattening it to form a plano-convex lens.

[0068] Specifically, an adsorbent 11 with a fine tip 13 is used to adsorb a transparent polymer 12, and the transparent polymer 12 is transferred into the central hole of the nylon sheet and naturally flattened to form a plano-convex lens 16. The adsorbent includes an acupuncture needle, an AFM probe or a semi-tapered optical fiber, and the transparent polymer includes polydimethylsiloxane or polymethyl methacrylate or UV-curing glue.

[0069] Please refer to FIG2 , which is a schematic diagram of a process for preparing a plano-convex lens on a glass sheet according to an embodiment of the present application, comprising a syringe 11 ; polydimethylsiloxane liquid 12 ; an acupuncture needle 13 ; a polydimethylsiloxane droplet 14 ; a glass disk 15 ; and a polydimethylsiloxane plano-convex lens 16 .

[0070] Step S113: thermally curing the glass sheet to form a solid plano-convex lens.

[0071] In this embodiment, the step of thermally curing the glass sheet to form a solid plano-convex lens specifically includes the following steps: placing the glass sheet in a dryer and thermally curing it to form a solid plano-convex lens, the curing temperature is 40°C-100°C, and the curing time is 20 minutes to 3 hours.

[0072] It can be understood that the thickness and curvature radius of the plano-convex lens can be controlled by controlling the amount of the transparent polymer and the size of the central hole of the nylon sheet. The process is controllable and the technology is simple.

[0073] Step S120: transferring the transparent polymer droplet to the center of the plano-convex lens.

[0074] In this embodiment, the step of transferring the transparent polymer droplet to the center of the plano-convex lens specifically includes the following steps S121 to S125 , and the implementation method of each step is described in detail below.

[0075] Step S121: Obtain two sections of semi-tapered optical fibers.

[0076] In this embodiment, the step of obtaining two sections of semi-tapered optical fibers specifically includes the following steps:

[0077] One end of a single-mode optical fiber with a coating stripped off is fixed to an optical rod, and the other end is fixed to a three-dimensional translation stage; the bare optical fiber with the coating stripped off is heated, and the translation stage knob is rotated to stretch the optical fiber to form a tapered optical fiber; the optical fiber is continued to be stretched using the translation stage until the optical fiber breaks to obtain two semi-tapered optical fibers. The heating can be performed using a butane torch, an arc lighter, an alcohol lamp, or a carbon dioxide laser.

[0078] Furthermore, the optical fiber in this embodiment can be glass optical fiber, plastic optical fiber, special glass optical fiber, etc.

[0079] Please refer to Figure 3, which illustrates the process of obtaining two semi-tapered optical fibers according to one embodiment. The diagram includes a fixing screw 1, an optical fiber support rod 2, an optical fiber clamp 3, a single-mode optical fiber 4, a bare optical fiber 5, an optical fiber clamp 6, a three-dimensional translation stage 7, a butane torch 8, a stretched tapered optical fiber 9, and a broken semi-tapered optical fiber 10. The detailed steps are described above and will not be repeated here.

[0080] Step S122: Add a small amount of transparent polymer droplets to a section of the semi-tapered optical fiber and allow it to slide from the tapered transition area to the tapered waist area under the action of gravity. The transparent polymer droplets attached to the tapered waist area of ​​the tapered optical fiber form tiny spindle-shaped droplets under the action of solid-liquid interfacial tension.

[0081] Step S123: Fixing the semi-tapered optical fiber with the spindle-shaped droplet attached to the three-dimensional translation stage.

[0082] Step S124: transferring one or more spindle-shaped droplets to the tip of the semi-tapered optical fiber fixed on another section of the three-dimensional translation stage by adsorption transfer to form a single tiny droplet.

[0083] Step S125: Transferring the tiny droplet to the center of the plano-convex lens via the three-dimensional displacement stage.

[0084] Please refer to Figure 4, which illustrates the process of transferring UV-curable adhesive to a plano-convex lens according to one embodiment of the present application. The process includes a glass slide 17; UV-curable adhesive liquid 18; a spindle-shaped droplet 19; a UV-curable adhesive droplet 20; and a UV-curable adhesive liquid film 21. The detailed steps are described above and will not be repeated here.

[0085] It should be noted that the fixture used to fix the optical fiber in this embodiment can also be replaced by other screws or clamps with fixing functions. The device used to stretch the optical fiber is not limited to an optical bracket or a translation stage, and other systems or devices with stretching functions can be used.

[0086] Step S130: fixing the microspheres to the transparent polymer droplets.

[0087] In this embodiment, the step of fixing the microspheres to the transparent polymer droplets specifically includes the following steps S131 to S135:

[0088] Step S131: dripping a suspension containing microspheres onto a glass slide, and then drying and heating to evaporate the liquid and leave the microspheres. The microspheres include hemispheres, spherical segments, spherical caps, and deformed microspheres. The microspheres are made of a high refractive index material, such as barium titanate glass, chalcogenide glass, or tellurite glass.

[0089] Step S132: Obtain a section of optical fiber, strip off the coating of the section of optical fiber at its end portion, fix the section of optical fiber to the glass slide and make the bare optical fiber with the coating stripped extend beyond the outer boundary of the glass slide, and then fix the glass slide to the three-dimensional translation stage.

[0090] Step S133: Manipulate the three-dimensional translation stage to make the tip of the bare optical fiber contact the single microsphere, and the microsphere will be adsorbed onto the tip of the bare optical fiber.

[0091] In this embodiment, the size of the microspheres is from several micrometers to dozens of micrometers.

[0092] Step S134: placing the glass sheet with the plano-convex lens under the observation field of view, manipulating the three-dimensional translation stage to position and transfer the microspheres onto the transparent polymer droplet.

[0093] Step S135: solidifying the glass sheet with the plano-convex lens and the microspheres, so that the microspheres are fixed to the transparent polymer droplets.

[0094] In this embodiment, the step of curing the glass sheet with the plano-convex lens and the microspheres so that the microspheres are fixed to the transparent polymer droplets specifically includes: placing the glass sheet with the plano-convex lens and the microspheres under ultraviolet light for irradiation and curing so that the microspheres are fixed to the transparent polymer droplets, wherein the irradiation time is 10-60 minutes, and the irradiation power range of the ultraviolet lamp is 30W-100W.

[0095] Please refer to Figure 5, which is a schematic diagram of the process of fixing the microspheres to the transparent polymer droplets provided in one embodiment of the present application, including a single-mode optical fiber 4; a bare optical fiber 5; a glass disk 15; a polydimethylsiloxane plano-convex lens 16; a glass slide 17; a UV-curable adhesive liquid film 21; and high-refractive-index microspheres 22. The detailed steps can be found in the above description and will not be repeated here.

[0096] Please refer to Figure 6, which is a top view of the plano-convex lens provided in this embodiment after microspheres are placed, with the arrows pointing to the microspheres. Please refer to Figure 7, which is a side view (left) of the polydimethylsiloxane plano-convex lens provided in this embodiment after microspheres are placed (right, with the arrows pointing to the microspheres).

[0097] Step S140: fixing the glass sheet with the microspheres fixed thereon to an objective lens to obtain the super-resolution microscope objective lens.

[0098] In this embodiment, the step of fixing the glass sheet with the microspheres fixed thereon to the objective lens to obtain the super-resolution microscope objective lens specifically includes the following steps S141 to S142. The implementation method of each step is described in detail below.

[0099] Step S141: fix the glass sheet with the microspheres fixed thereon to the ring 232 and heat and cure it. The glass sheet with the microspheres fixed thereon is fixed to the objective lens to obtain the super-resolution microscope objective lens.

[0100] In this embodiment, the step of fixing the glass sheet with the microspheres fixed to the ring and heating and curing it, and fixing the glass sheet with the microspheres to the objective lens to obtain the super-resolution microscope objective lens specifically includes the following steps: using a pipette to absorb a small amount of colloidal material and applying it on the ring, placing the glass sheet with the microspheres fixed on the ring, and placing it in a dryer for heating and curing, so that the glass sheet is fixed to the sleeve to obtain the super-resolution microscope objective lens.

[0101] Furthermore, the glue material used to fix the glass sheet in this embodiment may be photoresist or may be replaced by other glue materials that can be cured by heating.

[0102] Step S142: providing a sleeve.

[0103] Referring to Figures 8 to 9, the sleeve includes a fixed portion 26 and a rotating portion 23. The fixed portion 26 is nested and fixed on the objective lens 27. The rotating portion 23 is sleeved on the fixed portion 26 and can rotate around the fixed portion 26. The rotating portion 23 includes a hollow cylinder 231 and a ring 232 fixedly connected to the hollow cylinder 231. The inner wall of the hollow cylinder 231 is provided with a thread. By rotating the thread, the rotating portion 23 can be adjusted to move up and down along the fixed portion 26. The glass piece fixed with the microspheres is fixed to the ring 232 and heated and cured. Then, the fixed portion 26 of the sleeve is fixed to the objective lens. Then, the rotating portion 23 of the sleeve containing the ring 232 is fixed to the fixed portion 26 of the sleeve. That is, the sleeve including the glass piece with microspheres is fixed to the objective lens to obtain the super-resolution microscope objective lens.

[0104] Furthermore, the rotating part 23 further includes a support column 233 , and the support column 233 is fixedly connected to the ring 232 and the hollow cylinder 231 .

[0105] In this embodiment, the Teflon material used to make the sleeve can be replaced by other high temperature resistant and corrosion resistant materials. The objective lens includes a water immersion objective lens or an oil immersion objective lens.

[0106] Please refer to Figure 10, which illustrates the process for securing a glass slide with the microspheres fixed thereto to an objective lens to obtain the super-resolution microscope objective lens according to one embodiment of the present application. The process includes a rotating portion 23; a pipette 24; a photoresist 25; a fixed sleeve 26; and a water immersion objective lens 27. The detailed steps can be found in the above description and will not be repeated here.

[0107] The super-resolution microscope objective lens, manufacturing method, preparation method and super-resolution microscope device provided in the above embodiments of the present application are prepared in a controllable manner on a glass sheet to make a plano-convex lens, and then the microspheres are controllably positioned and fixed on the upper surface of the center position of the plano-convex lens by precise micro-manipulation and heat / light curing, and integrated with the sleeve and assembled to the objective lens to form a microsphere-assisted super-resolution microscope objective lens. The super-resolution microscope objective lens provided by the present application adds water or oil above the glass sheet without contacting the sample, and there is no potential contamination or impact. Compared with other methods of directly placing microspheres on a plane or a flat plate, the present application places the microspheres on the plano-convex lens, which can effectively avoid the problem that other places of the objective lens first contact the sample when the microspheres assist imaging, causing the microspheres to be unable to achieve imaging. At the same time, the plano-convex lens can be adjusted as needed, and the plano-convex lens has a certain enhancing effect on the imaging effect of the microspheres. In addition, the above-mentioned manufacturing method is simple and easy to control, easy to operate and implement, and can improve the performance of conventional optical microscopes at a low cost.

[0108] It can be understood that the various technical features of the above-described embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the various technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0109] The above are merely preferred embodiments of the present application and only specifically describe the technical principles of the present application. These descriptions are intended only to explain the principles of the present application and should not be construed in any way as limiting the scope of protection of the present application. Based on the explanations herein, any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present application, as well as other specific implementations of the present application that can be conceived by those skilled in the art without inventive effort, shall be included within the scope of protection of the present application.

Claims

1. A method for manufacturing a super-resolution microscope objective lens, characterized in that: The steps include: Prepare plano-convex lenses on glass slides; transferring a transparent polymer droplet to the center of the plano-convex lens; Fixing microspheres to the transparent polymer droplets; The glass sheet with the microspheres fixed thereon is fixed to an objective lens to obtain the super-resolution microscope objective lens.

2. The method for manufacturing a super-resolution microscope objective lens according to claim 1, characterized in that: The step of preparing a plano-convex lens on a glass sheet specifically includes the following steps: Providing a glass sheet, and placing a nylon sheet with a central opening on the glass sheet; Transferring a transparent polymer droplet into the central hole of the nylon sheet and naturally flattening it to form a plano-convex lens; The glass sheet is thermally cured to form a solid plano-convex lens.

3. The method for manufacturing a super-resolution microscope objective lens according to claim 2, characterized in that: The step of transferring the transparent polymer droplet into the central hole of the nylon sheet and naturally paving to form a plano-convex lens specifically includes the following steps: A transparent polymer is adsorbed by an adsorbent with a fine tip and transferred into the central hole of the nylon sheet, and naturally flattened to form a plano-convex lens. The adsorbent includes an acupuncture needle, an AFM probe or a semi-tapered optical fiber, and the transparent polymer includes polydimethylsiloxane or polymethyl methacrylate or UV-curing glue.

4. The method for manufacturing a super-resolution microscope objective lens according to claim 2, characterized in that: The step of thermally curing the glass sheet to form a solid plano-convex lens specifically includes the following steps: The glass sheet is placed in a drying machine and thermally cured to form a solid plano-convex lens. The curing temperature is 40° C.-100° C. and the curing time is 20 minutes to 3 hours.

5. The method for manufacturing a super-resolution microscope objective lens according to claim 3, characterized in that: The thickness and the curvature radius of the plano-convex lens are controlled by controlling the amount of the transparent polymer and the size of the central hole of the nylon sheet.

6. The method for manufacturing a super-resolution microscope objective lens according to claim 1, characterized in that: The step of transferring the transparent polymer droplet to the center position of the plano-convex lens specifically includes the following steps: Obtain two sections of semi-tapered optical fiber; A small amount of transparent polymer droplets are added to a section of the semi-conical optical fiber, and the droplets are allowed to slide from the tapered transition area to the tapered waist area under the action of gravity. The transparent polymer droplets attached to the tapered waist area of ​​the tapered optical fiber form tiny spindle-shaped droplets under the action of the solid-liquid interface tension. fixing the semi-tapered optical fiber with the spindle-shaped droplet attached to a three-dimensional translation stage; Transferring one or more spindle-shaped droplets to the tip of a semi-tapered optical fiber fixed to another section of the three-dimensional translation stage by adsorption transfer to form a single tiny droplet; The tiny droplet is transferred to the center position of the plano-convex lens by the three-dimensional translation stage.

7. The method for manufacturing a super-resolution microscope objective lens according to claim 6, characterized in that: The step of obtaining two sections of semi-tapered optical fiber specifically includes the following steps: One end of a single-mode optical fiber with a coating stripped off is fixed to an optical connecting rod, and the other end is fixed to a three-dimensional translation stage; the bare optical fiber with the coating stripped off is heated, and the translation stage knob is rotated to stretch the optical fiber to form a tapered optical fiber; the optical fiber is continued to be stretched by the translation stage until the optical fiber breaks to obtain two sections of semi-tapered optical fiber, and the heating can be performed by a butane blowtorch, an arc lighter, an alcohol lamp, or a carbon dioxide laser.

8. The method for manufacturing a super-resolution microscope objective lens according to claim 1, characterized in that: The step of fixing the microspheres to the transparent polymer droplets specifically includes the following steps: The suspension containing microspheres is dripped onto a glass slide, and then dried and heated to evaporate the liquid to leave the microspheres, wherein the microspheres include hemispheres, spherical segments, spherical caps, and deformed microspheres; the material of the microspheres is a material with a high refractive index, and the high refractive index material includes barium titanate glass, chalcogenide glass, or tellurite glass; Obtain a section of optical fiber, strip off the coating layer of the section of optical fiber at the end portion thereof, fix the section of optical fiber to the glass slide and make the bare optical fiber stripped of the coating layer extend beyond the outer boundary of the glass slide, and then fix the glass slide to the three-dimensional displacement stage; Manipulating the three-dimensional translation stage to make the tip of the bare optical fiber contact with a single microsphere, and the microsphere will be adsorbed onto the tip of the bare optical fiber; Placing a glass sheet with the plano-convex lens under the observation field of view, manipulating the three-dimensional displacement stage, positioning and transferring the microsphere onto the transparent polymer droplet; The glass sheet with the plano-convex lens and the microspheres is solidified so that the microspheres are fixed to the transparent polymer droplets.

9. The method for manufacturing a super-resolution microscope objective lens according to claim 8, characterized in that: In the step of manipulating the three-dimensional translation stage to make the tip of the bare optical fiber contact the single microsphere, the microsphere is adsorbed onto the tip of the bare optical fiber, and the size of the microsphere is several micrometers to tens of micrometers.

10. The method for manufacturing a super-resolution microscope objective lens according to claim 8, characterized in that: The step of curing the glass sheet with the plano-convex lens and the microspheres so that the microspheres are fixed to the transparent polymer droplets specifically includes: placing the glass sheet with the plano-convex lens and the microspheres under ultraviolet light for irradiation and curing so that the microspheres are fixed to the transparent polymer droplets, wherein the irradiation time is 10-60 minutes, and the irradiation power range of the ultraviolet lamp is 30W-100W.

11. The method for manufacturing a super-resolution microscope objective lens according to claim 1, characterized in that: The step of fixing the glass sheet with the microspheres fixed thereon to the objective lens to obtain the super-resolution microscope objective lens specifically includes the following steps: A sleeve is provided, the sleeve comprising a fixed part and a rotating part, the fixed part is nested and fixed on the objective lens, the rotating part is sleeved on the fixed part and can rotate around the fixed part, the rotating part comprises a hollow cylinder and the ring fixedly connected to the hollow cylinder, the inner wall of the hollow cylinder is provided with a thread, and the rotating part can be adjusted to move up and down along the fixed part by rotating the thread; The glass sheet with the microspheres fixed thereon is fixed to the circular ring and heated and cured, and then the fixed part of the sleeve is fixed to the objective lens, and then the rotating part of the sleeve including the circular ring is fixed to the fixed part of the sleeve, and the sleeve including the glass sheet with the microspheres is fixed to the objective lens to obtain the super-resolution microscope objective lens.

12. The method for manufacturing a super-resolution microscope objective lens according to claim 11, characterized in that: The rotating part also includes a supporting column, and the supporting column is fixedly connected to the ring and the hollow cylinder.

13. The method for manufacturing a super-resolution microscope objective lens according to claim 11, characterized in that: The step of fixing the glass sheet with the microspheres fixed thereon to the ring, heating and curing the glass sheet with the microspheres fixed to the objective lens to obtain the super-resolution microscope objective lens specifically includes the following steps: A small amount of colloidal material is sucked up with a pipette and applied on the ring, a glass sheet fixed with the microspheres is placed on the ring, and placed in a dryer for heating and curing, so that the glass sheet is fixed to the sleeve to obtain the super-resolution microscope objective lens.

14. The method for manufacturing a super-resolution microscope objective lens according to claim 13, characterized in that: The objective lens includes a water immersion objective lens or an oil immersion objective lens.

15. A super-resolution microscope objective lens, characterized in that: The method is prepared according to any one of claims 1 to 14.

16. A super-resolution microscopy device, characterized in that: Including the super-resolution microscope objective lens described in claim 15.

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