CN114153062A - Super-surface objective lens, focusing method thereof and fluorescence microscope - Google Patents
Super-surface objective lens, focusing method thereof and fluorescence microscope Download PDFInfo
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- CN114153062A CN114153062A CN202111485812.6A CN202111485812A CN114153062A CN 114153062 A CN114153062 A CN 114153062A CN 202111485812 A CN202111485812 A CN 202111485812A CN 114153062 A CN114153062 A CN 114153062A
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- G—PHYSICS
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- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B1/00—Optical elements characterised by the material of which they are made; Optical coatings for optical elements
- G02B1/002—Optical elements characterised by the material of which they are made; Optical coatings for optical elements made of materials engineered to provide properties not available in nature, e.g. metamaterials
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B1/00—Optical elements characterised by the material of which they are made; Optical coatings for optical elements
- G02B1/04—Optical elements characterised by the material of which they are made; Optical coatings for optical elements made of organic materials, e.g. plastics
- G02B1/041—Lenses
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
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Abstract
The invention relates to the field of optical elements, in particular to a super-surface objective lens, a focusing method thereof and a fluorescence microscope. The super-surface objective lens is provided with a super-surface structure; the super-surface structure comprises a plurality of repeating units and a substrate, wherein the repeating units are arranged on the substrate and are arranged periodically; the repeating unit is a sub-wavelength micro-nano structure and is used for generating plasma coupling resonance on the substrate; the substrate is flexible and dynamically tunable for controlling plasmon coupled resonance. The super-surface objective lens prepared by the invention has very small size, is beneficial to integration, and has the advantages of simple structure, easy steps and strong operability.
Description
Technical Field
The invention relates to the field of optical elements, in particular to a super-surface objective lens, a focusing method thereof and a fluorescence microscope.
Background
The fluorescence microscope uses ultraviolet as light source to irradiate the object to be detected and make it emit fluorescence, then under the microscope lens the shape of the object and its position can be observed.
The fluorescence microscope comprises a light source, a light filtering system, a reflector, a condenser, an objective lens, an ocular lens and an epi-illumination device. The objective is an important optical component in the microscope, and plays a first step of magnification on the object, thereby affecting the imaging quality and various optical technical parameters. The objective lens has a complex structure and is composed of a plurality of lens groups fixed in an objective lens barrel, each lens group is formed by gluing a series of lenses with different parameters, and the plurality of lens groups are matched with each other to effectively correct the aberration of the objective lens.
In practical application, the tunable focal length needs to be realized by moving the objective lens group, and the adjustment of the objective lens group changes the thickness of the objective lens group. However, the traditional objective lens group has large volume and complex structure.
Disclosure of Invention
The invention provides a super-surface objective lens, a focusing method thereof and a fluorescence microscope, which at least solve the technical problems in the prior art.
In a first aspect, the present application provides a super-surface objective lens, which adopts the following technical scheme: the super-surface objective lens is provided with a super-surface structure; the super-surface structure comprises a plurality of repeating units and a substrate, wherein the repeating units are arranged on the substrate and are arranged periodically; the repeating unit is a sub-wavelength micro-nano structure and is used for generating plasma coupling resonance on the substrate; the substrate is flexible and dynamically tunable for controlling plasmon coupled resonance.
Optionally, the material used for manufacturing the substrate is any one of polydimethylsiloxane, polymethyl methacrylate, polyamide, polyimide, polyethylene terephthalate, bis-p-chloromethylphenyl, polyimide, polypropylene, parylene, SU-8 photoresist, or polystyrene.
Optionally, the sub-wavelength micro-nano structure is a nano-pillar, and the shape of the nano-pillar is any one or more of a polygon, a cylinder and a cone.
Optionally, the nanopillars are made of a high index dielectric material. Further, the high-refractive-index dielectric material is any one of silicon nitride, chalcogenide glass, amorphous silicon, aluminum oxide, titanium dioxide and zinc oxide.
Optionally, the transmittance of the repeating unit is 50% or more. Further, the transmittance of the repeating unit is 80% or more.
In a second aspect, the present application provides a focusing method for a super-surface objective, comprising the following steps:
s1, performing parameter scanning and simulation on the repeating units, realizing phase coverage larger than 2 pi by changing the size of the repeating units, wherein the phase distribution of the repeating units satisfies the following formula:
where lambda is the wavelength of the incident light,fis the focal length of the objective lens, and r is the distance between the focal point and the center of the lens;
and S2, after the phase distribution in the S1 is obtained, simulating the super-surface objective lens to obtain the corresponding focusing effect condition, so that the emergent light of the super-surface objective lens is converged at the focal position.
In a third aspect, the present application provides a fluorescence microscope comprising any of the above-described super-surface objective lenses.
Optionally, the device further comprises an eyepiece, a light source, an object stage, a filter system and a lens barrel, wherein the filter system is arranged in the lens barrel; the filtering system comprises a spectroscope, an optical filter and a barrel mirror; the optical filter is used for filtering incident light from the light source, the spectroscope is used for reflecting the incident light from the light source, and the tube lens is used for filtering stray light entering the eyepiece; the light emitted from the light source sequentially passes through the optical filter, the spectroscope and the cylindrical lens, the spectroscope reflects exciting light with specific wavelength to the super-surface objective lens, and fluorescence excited by a specimen on the objective table is not reflected, so that the fluorescence passes through the ocular lens and is observed by human eyes.
Optionally, the light source is a high-pressure mercury lamp light source, and the illumination mode is an epi-type or a transmission type.
Compared with the prior art, the method has the following beneficial effects:
1. the subwavelength micro-nano structure repeating unit is used for generating plasma coupling resonance on the substrate to realize electric field regulation and control of incident light, the substrate is used for realizing dynamic regulation of a plasma coupling resonance phenomenon, and the super-surface objective prepared by the invention has very small size, is beneficial to integration, and has simple structure, easy steps and strong operability;
2. the super-surface structure has extensibility, and the adjustment of the surface plasma coupling peak position is realized by changing the distance between the resonance units through external stress;
3. because the repeating unit of the super-surface objective consists of the transparent nano-pillars and the bendable organic substrate, the repeating unit has the characteristics of high refractive index and low loss in a visible light range, and therefore, the super-surface objective has higher focusing efficiency;
4. according to the method, parameter scanning and simulation are carried out on the repeating units, phase covering larger than 2 pi can be achieved by changing the diameter of the nano-column, meanwhile, the phase distribution of the repeating units is required to meet the formula requirement, so that the phase fitting result is basically consistent with ideal distribution, the control error is within a range of 5 degrees, the transmissivity of the repeating units is enabled to be more than 50%, and the super-surface objective lens has the advantage of high efficiency.
Drawings
Fig. 1 is a schematic structural view of a super-surface structure in the present application.
FIG. 2 is a phase and transmittance profile of different diameter repeating units when the incident light is 520nm visible in the present application, with the abscissa representing the diameter of the nanopillar in the parameter scan, in the range of 50nm to 300 nm.
FIG. 3 is an ideal phase profile for a hypersurface objective of the present application, where each point represents a cylindrical repeating unit at one diameter parameter.
FIG. 4 is a graph of the actual unit phase distribution of the hypersurface objective of the present application, where each point represents a cylindrical repeating unit for one diameter parameter.
FIG. 5 is an electric field distribution diagram of the super-surface objective lens in the xz plane in the present application.
FIG. 6 is an electric field distribution diagram of the super-surface objective lens of the present application along the x-axis at the focal plane.
FIG. 7 is a schematic view of the optical path of a fluorescence microscope of the present application.
Description of reference numerals:
1. a super-surface objective lens; 11. a super-surface structure; 111. a repeating unit; 112. a substrate; 2. an eyepiece; 3. a light source; 4. an object stage; 51. a beam splitter; 52. an optical filter; 53. a cylindrical mirror; 7. a lens barrel.
Detailed Description
The invention is explained below with reference to the figures and examples, which are intended to illustrate the invention and are not to be construed as limiting the invention. While exemplary embodiments of the invention are shown in the drawings, it should be understood that the invention can be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
Referring to fig. 1, a super-surface objective 1 is provided with a super-surface structure 11; the super-surface structure 11 comprises a plurality of repeating units 111 and a substrate 112, wherein the plurality of repeating units 111 are arranged on the substrate 112 and are arranged periodically; the repeating unit 111 is a sub-wavelength micro-nano structure and is used for generating plasma coupling resonance on the substrate 112; substrate 112 is flexible and dynamically tunable for controlling plasmon coupled resonance.
The material used for forming the substrate 112 is any one of Polydimethylsiloxane (PDMS), polymethyl methacrylate, polyamide, polyimide, polyethylene terephthalate, bis-p-chloromethylphenyl, polyimide, polypropylene, parylene, SU-8 photoresist, or polystyrene. The materials have good chemical stability, air permeability, light transmittance, elasticity and biocompatibility, and can be well compatible with the traditional semiconductor micro-nano processing integration technology.
The sub-wavelength micro-nano structure is a nano column, and the nano column is a nano structure of a row-column lattice. The shape in fig. 1 is for convenience of explanation of the unit structure thereof, and is not the only shape. The shape of the nano-pillar can be any one or more of a polygon, a cylinder and a cone. In this embodiment, the nano-pillar is taken as an example for explanation, the period of the repeating unit 111 is 200nm to 800nm, the height of the nano-pillar is 100nm to 2000nm, and the diameter of the nano-pillar is 50nm to 700 nm.
The nano-pillars are made of high-refractive-index dielectric material, such as silicon nitride, chalcogenide glass, amorphous silicon, aluminum oxide, and titanium dioxide (TiO)2) And zinc oxide.
The transmittance of the repeating unit 111 is 50% or more, and phase coverage of more than 2 pi can be achieved by changing the diameter of the nanopillar. In this embodiment, a TiO2 nanorod and a PDMS substrate 112 are used as an example. From TiO2The super-surface structure 11 formed by the nano-pillars and the PDMS substrate 112 has the characteristics of high refractive index and low loss in the visible light range, so that the focusing efficiency is high.
The focusing method based on the super-surface objective lens comprises the following steps:
s1, carrying out parameter scanning and simulation on the repeating units by using FDTD-solutions software, wherein the transmittance of each repeating unit is required to reach more than 50%, and phase coverage larger than 2 pi is realized by changing the diameter of the nano-column;
the optimized parameters of the obtained repeating unit are that the period P =350nm, the height H =500nm of the nano-column, and the diameter variation range D = 50-350 nm. The phase and transmittance distribution of the repeating unit is shown in fig. 2; FIG. 2 is a phase and transmittance distribution graph of different diameter repeating units when the incident light is 520nm visible light, the abscissa represents the diameter of the nanopillar in the parameter scan, in the range of 50nm to 300 nm.
S2, according to the traditional optical theory, to converge the full aperture light to the same point, the phases of the incident lights with different apertures at the focus point must be the same, so that the ideal phase distribution of the aspheric surface objective lens must satisfy the following formula:
wherein lambda is the wavelength of incident light, f is the focal length of the objective lens, and r is the distance between the focal point and the center of the objective lens;
in the embodiment, a small-size convergent lens is designed and simulated, the diameter of the super-surface objective lens is about 35um, the focal length is 35um, and the design wavelength is 520nm of visible light;
fig. 3 and 4 are respectively the fitting result of the phase distribution and the fitting result of the transmittance distribution of the super-surface objective lens calculated according to the formula. FIG. 3 is an idealized phase profile of a super-surface objective of the present invention, where each point represents a cylindrical repeating unit at one diameter parameter. FIG. 4 is a graph of the phase distribution of an actual unit of the super-surface objective of the present invention, where each point represents a cylindrical repeating unit at one diameter parameter. It can be seen that the phase fitting result is basically consistent with the ideal distribution, the error is within the range of 5 degrees, the transmittance of the whole super-surface objective lens is distributed over 50 percent, and the super-surface objective lens has the advantage of high efficiency.
S3, after obtaining the ideal phase distribution of the super-surface objective lens, using FDTD-solutions software to simulate the super-surface objective lens, and obtaining the focusing effect of the super-surface objective lens as shown in fig. 5 and 6, which are respectively an xz plane electric field distribution diagram and a focal plane electric field distribution diagram.
FIG. 5 is an electric field distribution diagram of the super-surface objective lens of the present invention in the xz plane. Referring to fig. 5, the actual focus position obtained by simulation is z = 35.58um, which is substantially in accordance with the design value f =35 um, and the error is within one wavelength range.
FIG. 6 is a diagram of the electric field distribution along the x-axis at the focal plane of the super-surface objective of the present invention. Referring to fig. 6, it can be calculated that the focusing efficiency of the superlens reaches 80% or more.
Based on the focusing method of the super-surface objective lens, the super-surface objective lens is simulated by calculating the phase distribution of the repeating units to obtain the corresponding focusing effect condition, and the emergent light of the super-surface objective lens is converged at the focus position by changing the diameter of the nano-column to change the phase coverage, wherein the phase coverage is more than 2 pi.
FDTD-Solutions software is produced by Canada logical Solutions company, is based on solving of vector three-dimensional Maxwell dimensional equation, adopts a time domain finite difference FDTD method to grid space, calculates step by step in time, obtains steady-state continuous wave result of a wide band from a time domain signal, can accurately describe the dispersion characteristic of a material in the wide band by a unique material model, is embedded with a high-speed and high-performance calculation engine, can obtain the result of multiple wavelengths of the wide band by one-time calculation, can simulate any three-dimensional shape, and provides an accurate dispersion material model.
The fluorescence microscope, as shown in fig. 7, includes a super-surface objective 1, an eyepiece 2, a light source 3, a stage 4, a filter system 5 and a lens barrel 7, wherein the filter system 5 is disposed in the lens barrel 7, and the lens barrel 7 is T-shaped. The light source 3 is a high-pressure mercury lamp light source, and the illumination mode is an epi-type or a transmission type, and the epi-type is taken as an example in this embodiment. The filter system 5 includes a beam splitter 51, a filter 52 and a tube lens 53, the filter 52 is used for filtering the incident light from the light source 3, the tube lens 53 is used for filtering the stray light entering the eyepiece 2, the beam splitter 51 reflects the incident light from the light source 3, and the angle between the beam splitter 51 and the horizontal plane can be any angle of 30 ° to 50 °, for example 45 ° in this embodiment. The spectroscope 51 can be arranged in the middle of the lens barrel 7, light emitted from the light source 3 passes through the optical filter 52, the spectroscope 51 and the barrel lens 53 in sequence, exciting light is emitted to the spectroscope 51, the spectroscope 51 reflects the exciting light with a specific wavelength to the super-surface objective lens 1, fluorescence excited by the specimen is not reflected, and the fluorescence passes through the ocular lens 2 to be observed by human eyes.
The implementation principle of the fluorescence microscope based on the super-surface objective lens is as follows: in the using process, a sample is firstly placed on the sample table and fixed, the light source 3 is turned on and preheated for a certain time, and observation is carried out after the light source 3 emits light and tends to be stable. The excitation light is adjusted to proper intensity by adjusting the intensity of the light source 3, the position of the lens barrel 7 is adjusted up and down, and the stress applied to the super-surface objective lens 1 is synchronously adjusted to realize focusing alignment, so that a clear fluorescent image is obtained. The adjustment stress can be obtained by a uniform force application in the form of a shrink ring or the like.
It is to be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in the process, method, article, or apparatus that comprises the element.
The above are merely examples of the present application and are not intended to limit the present application. Various modifications and changes may occur to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.
Claims (10)
1. A super-surface objective lens, comprising: the super-surface objective lens is provided with a super-surface structure;
the super-surface structure comprises a plurality of repeating units and a substrate, wherein the repeating units are arranged on the substrate and are arranged periodically;
the repeating unit is a sub-wavelength micro-nano structure and is used for generating plasma coupling resonance on the substrate;
the substrate is flexible and dynamically tunable for controlling plasmon coupled resonance.
2. A super-surface objective lens according to claim 1, characterized in that: the material used for preparing the substrate is any one of polydimethylsiloxane, polymethyl methacrylate, polyamide, polyimide, polyethylene terephthalate, bis-p-chloromethyl benzene, polyimide, polypropylene, parylene, SU-8 photoresist or polystyrene.
3. A super-surface objective lens according to claim 1, characterized in that: the sub-wavelength micro-nano structure is a nano column, and the shape of the nano column is any one or more of a polygon, a cylinder and a cone.
4. A super-surface objective lens according to claim 3, characterized in that: the nanopillars are made of a high index of refraction dielectric material.
5. The super-surface objective lens according to claim 4, wherein: the high-refractive-index dielectric material is any one of silicon nitride, chalcogenide glass, amorphous silicon, aluminum oxide, titanium dioxide and zinc oxide.
6. A super-surface objective lens according to claim 1, characterized in that: the transmittance of the repeating unit is 50% or more.
7. The focusing method of the super surface objective lens as claimed in any one of claims 1 to 6, comprising the steps of:
s1, performing parameter scanning and simulation on the repeating units, realizing phase coverage larger than 2 pi by changing the size of the repeating units, wherein the phase distribution of the repeating units satisfies the following formula:
where lambda is the wavelength of the incident light,fis the focal length of the objective lens, and r is the distance between the focal point and the center of the lens;
and S2, after the phase distribution in the S1 is obtained, simulating the super-surface objective lens to obtain the corresponding focusing effect condition, so that the emergent light of the super-surface objective lens is converged at the focal position.
8. A fluorescence microscope, characterized by: comprising the hyper-surface objective of any one of claims 1 to 7.
9. A fluorescence microscope according to claim 8, wherein: the device also comprises an eyepiece, a light source, an objective table, a filtering system and a lens cone, wherein the filtering system is arranged in the lens cone;
the filtering system comprises a spectroscope, an optical filter and a barrel mirror;
the optical filter is used for filtering incident light from the light source, the spectroscope is used for reflecting the incident light from the light source, and the tube lens is used for filtering stray light entering the eyepiece;
the light emitted from the light source sequentially passes through the optical filter, the spectroscope and the cylindrical lens, the spectroscope reflects exciting light with specific wavelength to the super-surface objective lens, and fluorescence excited by a specimen on the objective table is not reflected, so that the fluorescence passes through the ocular lens and is observed by human eyes.
10. A fluorescence microscope according to claim 9, wherein the light source is a high pressure mercury lamp light source and the illumination mode is epi-illumination or transmission.
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