Compound lens optical super-resolution imaging system

Abstract

The composite lens optical super-resolution imaging system provided in this application fixes a microsphere lens on a three-dimensional nano-displacement stage unit, while simultaneously coating a thin film on the sample surface. A hemispherical lens, which can naturally form a hemisphere on the thin film due to the surface tension of the droplet, is transferred onto the thin film. When the three-dimensional nano-displacement stage unit moves the microsphere lens to the vertical direction of the hemispherical lens above the thin film, the hemispherical lens achieves primary super-resolution imaging when light illuminates the sample surface. Then, secondary magnification is achieved through the microsphere lens, and the image is transmitted to the microscope objective, thereby achieving the goal of breaking through the diffraction limit and improving the resolution and magnification of the optical microscope.

Description

Technical Field

[0001] This application relates to the field of optical device technology, and in particular to a composite lens optical super-resolution imaging system. Background Technology

[0002] Optical microscopes play a crucial role in numerous fields. However, the continuous development of micro-nano science and biomedicine has placed higher demands on high-resolution microscopic imaging technology. Limited by the optical diffraction limit, the resolution of conventional optical microscopes is difficult to exceed 200 nm. Microsphere lens-assisted super-resolution microscopy is a simple and easy method to solve this problem. After the sample is illuminated, near-field high spatial frequency evanescent waves carrying information about the fine structure of the nanoscale sample are coupled by the microspheres. The microspheres then convert this information to the far field, where it is received by the objective lens and propagated to the detector for display. This method has advantages such as no need for fluorescent labeling, direct imaging, simple component construction, and low cost. Super-resolution imaging is achieved through a simple combination of microsphere lenses and optical microscopes. However, in some existing microsphere lens-assisted super-resolution imaging technologies, the hemispherical lenses used have limited magnification during imaging, and the image cannot be observed with low-magnification objectives. Therefore, high-magnification objectives must be used. However, high-magnification objectives have short focal lengths and small fields of view, limiting the application space of super-resolution imaging based on different magnification objectives. Summary of the Invention

[0003] Therefore, it is necessary to provide a compound lens optical super-resolution imaging system that expands the application space of super-resolution imaging based on objectives of different magnifications, addressing the shortcomings of existing technologies.

[0004] To solve the above problems, this application adopts the following technical solution:

[0005] This application provides a composite lens optical super-resolution imaging system, comprising: a three-dimensional nanostage unit, a microsphere probe unit, a hemispherical lens unit, an optical microscope, and a computer, wherein:

[0006] The microsphere probe unit includes a microsphere lens, and the three-dimensional nanostage unit can move and control the position of the microsphere lens in the x, y, and z coordinate axes; the hemispherical lens unit includes a thin film coated on the sample to be tested and a hemispherical lens formed on the thin film; the optical microscope includes a stage on which the sample to be tested is placed, the optical microscope is used to receive super-resolution information from the hemispherical lens and the microsphere lens, and the computer is used to receive and process the super-resolution information.

[0007] In some embodiments, the three-dimensional nano-displacement stage unit includes a three-dimensional nano-displacement stage, a connecting rod fixed to the three-dimensional nano-displacement stage, an optical adjustment frame fixedly connected to the connecting rod, and a support fixedly connected to the optical adjustment frame. The three-dimensional nano-translation stage is a triaxial flexible displacement stage.

[0008] In some embodiments, the microsphere probe unit further includes a metal handle, an optical fiber handle, or a probe, the microsphere lens is fixed to the metal handle, optical fiber handle, or probe, and the metal handle, optical fiber handle, or probe is fixedly connected to the support.

[0009] In some embodiments, the microsphere lens is bonded to the metal handle, fiber optic handle, or probe by an adhesive.

[0010] In some embodiments, the microsphere lens may use barium titanate or arsenic sulfide microspheres when immersed in liquid, and may use silica or polystyrene microspheres when not immersed.

[0011] In some embodiments, the microsphere lens may also be fixed to the tip of a micropipette, embedded in a polymer film, or fixed to a microscope objective.

[0012] In some embodiments, the film is a curable polymer adhesive spin-coated onto the sample to be tested, the curable polymer adhesive including polydimethylsiloxane adhesive.

[0013] In some embodiments, the hemispherical lens may be a hydrophobic material or a hemisphere formed by cutting the microsphere lens.

[0014] In some embodiments, the optical microscope further includes a stage longitudinal movement knob and a stage lateral movement knob disposed below the stage, a stage coarse adjustment knob and a stage fine adjustment knob disposed on the side of the stage, a microscope objective, an objective turret connected to the microscope objective, an observation head fixedly connected to the objective turret, an eyepiece fixed to the observation head, a microscope tube fixed to the observation head, and a charge-coupled device (CCD) connected to the microscope tube.

[0015] In some embodiments, the computer is electrically connected to the charge-coupled device (CCD).

[0016] The present application adopts the above technical solution, and its beneficial effects are as follows:

[0017] The composite lens optical super-resolution imaging system provided in this application fixes a microsphere lens on a three-dimensional nano-displacement stage unit, while simultaneously coating a thin film on the sample surface. A hemispherical lens, which can naturally form a hemisphere on the thin film due to the surface tension of the droplet, is transferred onto the thin film. When the three-dimensional nano-displacement stage unit moves the microsphere lens to the vertical direction of the hemispherical lens above the thin film, the hemispherical lens achieves primary super-resolution imaging when light illuminates the sample surface. Then, secondary magnification is achieved through the microsphere lens, and the image is transmitted to the microscope objective, thereby achieving the goal of breaking through the diffraction limit and improving the resolution and magnification of the optical microscope. Attached Figure Description

[0018] To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the description of the embodiments of this application or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is a schematic diagram of the structure of the composite lens optical super-resolution imaging system provided in Embodiment 1 of this application.

[0020] Figure 2 This is a schematic diagram of the structure of the three-dimensional nano-displacement stage unit provided in Embodiment 1 of this application.

[0021] Figure 3 This is a schematic diagram of the structure of the microsphere probe unit provided in Embodiment 1 of this application.

[0022] Figure 4 This is a front view of the hemispherical lens unit provided in Embodiment 1 of this application.

[0023] Figure 5 This is a top view of the hemispherical lens unit provided in Embodiment 1 of this application. Detailed Implementation

[0024] The embodiments of this application are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain this application, and should not be construed as limiting this application.

[0025] In the description of this application, it should be understood that the terms "upper", "lower", "horizontal", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application.

[0026] 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 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 this application, "multiple" means two or more, unless otherwise explicitly specified.

[0027] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments.

[0028] Example 1

[0029] Please see Figure 1 This is a schematic diagram of the structure of a composite lens optical super-resolution imaging system provided in Embodiment 1, including: a three-dimensional nano-displacement stage unit 10, a microsphere probe unit 20, a hemispherical lens unit 30, an optical microscope 40, and a computer 50. The microsphere probe unit 10 includes a microsphere lens 21, and the three-dimensional nano-displacement stage unit 20 can move and control the position of the microsphere lens 21 along the x, y, and z coordinate axes. The hemispherical lens unit 30 includes a thin film 32 coated on the sample 31 to be tested and a hemispherical lens 33 formed on the thin film 32. The optical microscope 40 includes a stage 41 on which the sample 31 to be tested is disposed. The optical microscope 40 is used to receive the super-resolution information from the hemispherical lens and the microsphere lens. The computer 50 is used to receive and process the sample image. The connection relationships between the various components and their specific implementation methods are described in detail below.

[0030] Please see Figure 2 The three-dimensional nano-displacement stage unit 10 includes a three-dimensional nano-displacement stage 11, a connecting rod 12 fixed on the three-dimensional nano-displacement stage 11, an optical adjustment frame 13 fixedly connected to the connecting rod 12, and a support 14 fixedly connected to the optical adjustment frame 13. The three-dimensional nano-translation stage 11 is a triaxial flexible displacement stage, and the support 14 can be a right-angle support or a support with an inclined angle.

[0031] It is understood that the three-dimensional nano-translation stage 11 can be used to move and control the position of the microsphere lens 11 in the x, y, and z coordinate axes, the optical adjustment frame 13 is used to adjust the pitch and tilt angles of the microsphere lens 11, and the bracket 14 is used to rigidly connect the microsphere lens 11 and the optical adjustment frame 13.

[0032] Please see Figure 3 The microsphere probe unit 20 includes a microsphere lens 21 and a metal handle 22 (or an optical fiber handle or a probe). The microsphere lens 21 is fixed to the metal handle 22, and the metal handle is fixedly connected to the support 14.

[0033] Furthermore, the microsphere lens 21 is bonded to the metal handle 22 by an adhesive. The adhesive can be any substance that has adhesive properties and is curable.

[0034] Furthermore, the microsphere lens 21 can use barium titanate or arsenic sulfide microspheres when immersed in liquid, and can use silicon dioxide or polystyrene microspheres when not immersed.

[0035] Furthermore, the microsphere lens 21 can also be fixed to the tip of a micropipette, embedded in a polymer film, or fixed to a microscope objective.

[0036] Please see Figure 4 and Figure 5 The hemispherical lens unit 30 includes a thin film 32 coated on the sample 31 to be tested and a hemispherical lens 33 formed on the thin film 32.

[0037] Furthermore, the film 32 is a curable polymer adhesive spin-coated onto the sample to be tested, and the curable polymer adhesive includes polydimethylsiloxane adhesive.

[0038] Furthermore, the hemispherical lens 33 can be made of a hydrophobic material or a hemisphere formed by cutting the microsphere lens 33.

[0039] Please refer to the following: Figure 1 The optical microscope 40 also includes a stage longitudinal movement knob 42 and a stage lateral movement knob 43 disposed below the stage 41, a stage coarse adjustment knob 44 and a stage fine adjustment knob 45 disposed on the side of the stage 41, a microscope objective 46, an objective turret 47 connected to the microscope objective 46, an observation head 48 fixedly connected to the objective turret 47, an eyepiece 49 fixed to the observation head 48, a microscope tube 410 fixed to the observation head 48, and a charge-coupled device (CCD) 412 connected to the microscope tube 410. The computer electrical 50 is connected to the charge-coupled device (CCD) 412.

[0040] The composite lens optical super-resolution imaging system provided in Embodiment 1 of this application fixes a microsphere lens on a three-dimensional nano-displacement stage unit, while simultaneously coating a thin film on the sample surface. A hemispherical lens, which can naturally form a hemisphere on the thin film due to the surface tension of the droplet, is transferred onto the thin film. When the three-dimensional nano-displacement stage unit moves the microsphere lens to the vertical direction of the hemispherical lens above the thin film, the hemispherical lens achieves primary super-resolution imaging when light irradiates the sample surface. Then, secondary magnification is achieved through the microsphere lens, and the image is transmitted to the microscope objective, thereby achieving the purpose of breaking through the diffraction limit and improving the resolution and magnification of the optical microscope.

[0041] Example 2

[0042] The composite lens optical super-resolution imaging system provided in Embodiment 1 of this application has the following specific imaging method:

[0043] S1. The three-dimensional nano-stage 11 indirectly controls the position of the optical adjustment frame 13 and the support 14 via the connecting rod 12. The support 14 is fixed to the optical adjustment frame 13 using screws 15. The three-dimensional nano-stage 11 is a three-axis flexible displacement stage with a maximum stroke of 4mm per axis, a coarse adjustment stroke of 4mm, a fine adjustment stroke of 300μm, and a fine adjustment resolution of 100nm. The optical adjustment frame 13 is a compact optical adjustment frame with M4 mounting screw holes and an angle range of ±4°; the screws 15 are M4 stainless steel cap screws; the support 14 is a 2mm thick "L-shaped" aluminum support with a long side length of 160mm and a short side length of 14mm.

[0044] S2. The metal handle 22 and the microsphere lens 21 in the microsphere probe are bonded together with an adhesive, and the metal handle 22 is fixed to the support 14 with tape 23. The sample 31 to be tested, which has been spin-coated with a curable polymer film 32, and the hemispherical lens 33, which is naturally formed by surface tension and transferred onto the film using an optical fiber, are cured and then the three-dimensional nano-moving platform 11 is used to make the microsphere lens 21 and the hemispherical lens 33 coincide in the vertical direction.

[0045] S3, sample 31, thin film 32, and hemispherical lens 33 are placed on stage 41. The stage 41 and sample 31 are moved synchronously in the horizontal direction by the stage longitudinal movement knob 42 and stage lateral movement knob 43. The stage height coarse adjustment knob 44 and stage height fine adjustment knob 45 adjust the height of stage 41 and sample 31. The adjustment range of the stage height coarse adjustment knob 44 is approximately 24 mm, with a change of approximately 2 mm per revolution. The stage height fine adjustment knob 45 changes the stage height by approximately 200 μm per revolution.

[0046] Microscope objectives 46 with different magnifications and numerical apertures such as S4, 5×, 10×, 40×, 63× and 100× are mounted on the objective lens converter 47. The microscope objectives 46 are used to receive super-resolution images of the sample 31, which is smaller than the diffraction limit scale, after primary magnification by the hemispherical lens 33 and secondary magnification by the microsphere lens 21.

[0047] S5. The formed image is transmitted to the eyepiece 49 via the objective lens 46, or a digital format image is obtained using the charge-coupled device CCD detector 412 at the upper end of the lens tube 410 and the computer 50.

[0048] The composite lens optical super-resolution imaging method provided in Embodiment 2 of this application fixes a microsphere lens on a three-dimensional nano-displacement stage unit, while simultaneously coating a thin film on the sample surface. A hemispherical lens, which naturally forms a hemisphere on the film due to the surface tension of a droplet, is then transferred onto the film. When the three-dimensional nano-displacement stage unit moves the microsphere lens to the vertical direction of the hemispherical lens above the film, the hemispherical lens achieves primary super-resolution imaging when light illuminates the sample surface. Secondary magnification is then achieved through the microsphere lens, transmitting the image to the microscope objective. This method overcomes the diffraction limit and improves the resolution and magnification of the optical microscope. Furthermore, the above operation method is simple, the system is easy to implement, and it can improve the performance of an optical microscope with relatively low cost.

[0049] It is understood that the technical features of the above embodiments can be combined arbitrarily. For the sake of brevity, not all possible combinations of the technical features in the above 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.

[0050] The above are merely preferred embodiments of this application, and only specifically describe the technical principles of this application. These descriptions are only for explaining the principles of this application and should not be construed as limiting the scope of protection of this application in any way. Based on this explanation, any modifications, equivalent substitutions, and improvements made within the spirit and principles of this application, as well as other specific embodiments of this application that can be conceived by those skilled in the art without creative effort, should be included within the scope of protection of this application.

Claims

1. A composite lens optical super-resolution imaging system, characterized in that, The application relates to a three-dimensional nanometer displacement table unit, a microsphere probe unit, a hemispherical lens unit, an optical microscope and a computer, wherein: The microsphere probe unit comprises a microsphere lens, the three-dimensional nanometer displacement table unit can move and control the position of the microsphere lens in x, y and z three coordinate axes, the hemispherical lens unit comprises a film coated on a sample to be measured and a hemispherical lens formed on the film, the optical microscope comprises a stage, the sample to be measured is arranged on the stage, the optical microscope is used for receiving super-resolution information of the hemispherical lens and the microsphere lens, and the computer is used for receiving and processing the super-resolution information. The three-dimensional nanometer displacement table unit comprises a three-dimensional nanometer displacement table, a connecting rod fixed on the three-dimensional nanometer displacement table, an optical adjusting frame fixedly connected with the connecting rod and a support fixedly connected with the optical adjusting frame, and the three-dimensional nanometer displacement table is a three-axis flexible displacement table. The microsphere probe unit further comprises a metal handle or a fiber handle or a probe, the microsphere lens is fixed on the metal handle or the fiber handle or the probe, and the metal handle or the fiber handle or the probe is fixedly connected with the support. The microsphere lens and the hemispherical lens are coaxial in a vertical direction. The film is a curable polymer glue which is spin-coated on the sample to be measured, and the curable polymer glue comprises polydimethylsiloxane glue. The hemispherical lens is a hydrophobic substance, and the hemispherical lens is naturally formed on the film due to liquid drop surface tension. The microsphere lens is bonded on the metal handle or the fiber handle or the probe through an adhesive.

2. The compound lens optical super-resolution imaging system of claim 1, wherein, When the microsphere lens is immersed in liquid, barium titanate or arsenic sulfide microspheres can be used, and when the microsphere lens is not immersed, silica or polystyrene microspheres can be used.

3. The compound lens optical super-resolution imaging system of claim 1, wherein, The microsphere lens can also be fixed on a micropipette tip, embedded in a polymer film or fixed on a microscope objective.

4. The compound lens optical super-resolution imaging system of claim 1, wherein, The optical microscope further comprises a stage longitudinal movement knob and a stage transverse movement knob arranged below the stage, a height coarse adjustment knob and a stage height fine adjustment knob arranged on the side of the stage, a microscope objective, an objective converter connected with the microscope objective, an observation head fixedly connected with the objective converter, an eyepiece fixedly connected with the observation head, a lens barrel fixedly connected with the observation head and a charge coupled device (CCD) connected with the lens barrel.

5. The compound lens optical super-resolution imaging system of claim 1, wherein, The computer is electrically connected with the charge coupled device (CCD).

6. The compound lens optical super-resolution imaging system of claim 5, wherein, ​

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