CN112505009A - Super surface lens and fluorescence signal collection system formed by same - Google Patents
Super surface lens and fluorescence signal collection system formed by same Download PDFInfo
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- CN112505009A CN112505009A CN202011261855.1A CN202011261855A CN112505009A CN 112505009 A CN112505009 A CN 112505009A CN 202011261855 A CN202011261855 A CN 202011261855A CN 112505009 A CN112505009 A CN 112505009A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B3/00—Simple or compound lenses
- G02B3/10—Bifocal lenses; Multifocal lenses
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
Abstract
The invention provides a super-surface lens and a fluorescence signal collection system formed by the super-surface lens, wherein the super-surface lens comprises a substrate and a medium layer prepared on the substrate, a unit structure array is engraved on the medium layer and is used for focusing incident light with different wavelengths in an incident light wave band on different focal positions on the same focal plane, and the distance between two adjacent focal points is more than 2 mu m. The invention can reduce the complexity and the manufacturing difficulty of the fluorescence signal collecting system.
Description
Technical Field
The invention relates to the technical field of nucleic acid sequencing, in particular to a super-surface lens and a fluorescence signal collection system formed by the super-surface lens.
Background
Although the second-generation sequencing technology can obtain a large amount of sequencing data at low cost, due to the limitation of reading length, a large amount of sequencing fragments need to be correctly spliced to construct a genome map, and the requirements of functional genomics research on gene identification, cloning, gene structure, function and mutual relation thereof, gene expression regulation, protein structure identification, function and interaction and the like cannot be met.
The long read length characteristic of single-molecule fluorescent signal sequencing provides conditions for the splicing of repeated sequences of a genome. The single-molecule fluorescent signal sequencing technology is a technology for synthesizing and sequencing single DNA molecules based on fluorescent signal markers. Typical third-generation sequencing methods include exonuclease sequencing, sequencing by synthesis, nanopore sequencing, FRET transmission electron microscopy, and the like.
At present, the nucleic acid sequencing technology tends to be on chip and integrated. The nucleic acid sequencing technology based on fluorescence signal excitation has two main problems, namely effective excitation of a single-molecule fluorescence signal and guarantee of correct excitation of the signal in the sequencing process; and secondly, collection of single-molecule fluorescence signals ensures that the fluorescence signals can be effectively detected. In nucleic acid sequencing, a local optical field is generally used for exciting a fluorescence signal, and effective optical field local can ensure that only a single molecule exists in an excitation area and ordered signal excitation is ensured. Meanwhile, when signals are collected, because the strength of the excited fluorescence signals is low and the divergence angle is large, the fluorescence signals are collected by a lens with a large numerical aperture and are distinguished by combining an optical filter. At present, a common fluorescence signal collecting system collects fluorescence signal information in a mode of combining a Fresnel lens with an optical filter, the Fresnel lens can focus different wavelengths on the same focus, and the optical filter can select an excited fluorescence signal to realize the distinguishing of sequencing molecules. However, this method has high complexity in structure, great difficulty in process, and high manufacturing cost.
Disclosure of Invention
Aiming at the problems of high complexity and high manufacturing difficulty of the existing fluorescence signal collecting system, the invention provides a super-surface lens and a fluorescence signal collecting system formed by the super-surface lens, which can reduce the complexity and the manufacturing difficulty of the fluorescence signal collecting system.
In order to achieve the purpose, the invention adopts the following specific technical scheme:
the invention provides a super-surface lens which comprises a substrate and a medium layer prepared on the substrate, wherein a unit structure array is engraved on the medium layer and used for focusing incident light with different wavelengths in an incident light waveband at different focus positions on the same focal plane, and the distance between two adjacent focuses is larger than 2 mu m.
Preferably, the numerical aperture of the super-surface lens is greater than 0.4.
Preferably, the substrate is a silicon dioxide material, and the dielectric layer is a titanium dioxide material, a gallium nitride material or a hafnium dioxide material.
Preferably, the shape of the unit structures in the unit structure array is rectangular, cylindrical or elliptic cylindrical.
The invention also provides a fluorescence signal collecting system which comprises the super-surface lens.
Preferably, the fluorescence signal collecting system further comprises a nano structure, a waveguide structure, a transmission layer and a detector, wherein the waveguide structure is prepared on the substrate of the super surface lens, the nano structure is prepared on the waveguide structure, the transmission layer is prepared on the unit structure array of the super surface lens, and the detector is prepared on the transmission layer.
Preferably, the nanostructure is a zero mode waveguide structure or a nanophotonic ejection structure.
Preferably, the detector is a CMOS or SCMOS detector.
Preferably, the transmission layer is a silicon dioxide layer, a hafnium dioxide layer or a titanium dioxide layer.
The invention can obtain the following technical effects:
1. the super-surface lens has the characteristics of large numerical aperture, no polarization requirement on incident light and different focal point distribution positions;
2. the fluorescent signal collecting system can effectively collect the excited fluorescent signals in the gene sequencing process of the single-molecule fluorescent signals, ensures higher signal-to-noise ratio, and can distinguish focus positions corresponding to different wavelengths through the super-surface lens to replace an optical filter, thereby reducing the complexity and the manufacturing difficulty of the system.
Drawings
FIG. 1 is a schematic structural diagram of a super-surface lens according to a first embodiment of the invention;
FIG. 2 is a schematic structural diagram of a fluorescence signal collecting system according to a second embodiment of the present invention.
Wherein the reference numerals include: the device comprises a substrate 1, a unit structure 2, a nano structure 3, a waveguide structure 4, a transmission layer 5, a detector 6, a fluorescence signal 7 and a focal plane 8.
Detailed Description
Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. In the following description, the same reference numerals are used for the same blocks. In the case of the same reference numerals, their names and functions are also the same. Therefore, detailed description thereof will not be repeated.
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not to be construed as limiting the invention.
Example one
The embodiment of the invention provides a medium-based super-surface lens, which is used for realizing effective collection of a fluorescence signal.
Because the intensity of the fluorescence signal is low and the divergence angle is large, a super-surface lens with a larger numerical aperture is needed for realizing the collection of the fluorescence signal. In order to distinguish different fluorescence signals, the fluorescence signals with different wavelengths need to be focused at different focus positions. Based on the above two requirements, the super-surface lens needs to have characteristics of large numerical aperture and multiple focuses.
Based on the characteristics of the super-surface lens, the specific design of the super-surface lens is as follows:
1. and a super-surface lens with a numerical aperture larger than 0.4 and a large numerical aperture is selected to collect the fluorescence signal so as to meet the requirements of large divergence angle and low signal intensity of the fluorescence signal.
The diameter and focal length of the super-surface lens are selected according to the numerical aperture. For example, if the diameter of the super-surface lens is 10 μm and the focal length is 12 μm, the numerical aperture NA is 1.45 sin θ and 0.38 is 0.55, which ensures that the fluorescence signal can be collected efficiently.
2. The size and shape of the unit structure are designed to correspond to the fluorescent signals with different wavelengths, so that the fluorescent signals with different wavelengths are focused at different focus positions of the same focal plane.
More specifically, phase distribution of the super-surface lens is designed according to focus positions corresponding to different fluorescence signal wavelengths; according to the phase distribution, the unit structures are arranged, and the purpose that the focuses are at different positions of the same focal plane under different fluorescent signal wavelengths is achieved.
Fig. 1 shows a super-surface lens structure according to a first embodiment of the invention.
As shown in fig. 1, the super-surface lens includes a substrate 1 and a plurality of unit structures 2, the number of the unit structures 2 is multiple, a unit structure array is formed, the unit structures 2 are formed by photoetching a dielectric layer, specifically, a dielectric layer is prepared on the substrate 1, and the dielectric layer is subjected to patterned photoetching by an electron beam lithography technology to form a unit structure array formed by the plurality of unit structures 2.
The fluorescent signals 7 of different wavelengths are focused by the respectively corresponding unit structures 2 to different focal positions of the same focal plane 8. The super-surface lens can realize the fluorescent signal focusing function of the Fresnel lens, and compared with the Fresnel lens, the super-surface lens has the advantages of being simpler in structure, low in complexity, simple in manufacturing process and low in manufacturing cost.
In one example of the present invention, the substrate 1 may be silicon dioxide, and the dielectric layer may be titanium dioxide, gallium nitride, or hafnium dioxide.
One unit structure 2 can focus the fluorescent signal with one wavelength on one focal point, and the unit structure array can focus the fluorescent signals with different wavelengths on different focal positions of the same focal plane.
For example, the silicon dioxide substrate and the titanium dioxide medium are selected as working wavelengths, wherein the working wavelengths are 532nm, 578nm, 645nm and 721 nm. According to the four working wavelengths, four unit structures capable of responding to the four working wavelengths are designed, for example, the unit structure corresponding to the 532nm wavelength is rectangular, the unit structure corresponding to the 532nm wavelength is cylindrical, the unit structure corresponding to the 645nm wavelength is elliptic cylindrical, and the unit structure corresponding to the 645nm wavelength is triangular. And then arranging four unit structures of rectangle, cylinder, elliptic cylinder and triangle according to the focus positions corresponding to the wavelengths of 532nm, 578nm, 645nm and 721nm to realize the focusing function of the super-surface lens.
Example two
FIG. 2 shows a configuration of a fluorescence signal collecting system according to a second embodiment of the present invention.
As shown in fig. 2, the fluorescence signal collecting system includes a nano structure 3, a waveguide structure 4, a super-surface lens 1, a transmission layer 5 and a detector 6, which are sequentially prepared from top to bottom, wherein the waveguide structure 4 is prepared on a substrate of the super-surface lens 1 through a sputtering process or an atomic deposition process, the nano structure 3 is prepared on the waveguide structure 4 through the sputtering process or the atomic deposition process, the transmission layer 5 is prepared on a unit structure array of the super-surface lens 1 through the sputtering process or the atomic deposition process, and the detector 6 is prepared on the transmission layer 5.
The waveguide structure 4 is used to transmit excitation light to the nanostructures 3 to excite the fluorescent signal. The waveguide structure 4 comprises a core layer and a cladding layer, the cladding layer is wrapped outside the core layer, and the refractive index of the core layer is higher than that of the cladding layer.
The nano-structure 3 is used for exciting the nucleic acid molecule by exciting light, so that the nucleic acid molecule emits a fluorescent signal. The nano structure 3 can be a zero-mode waveguide structure or a nano-photon injection structure, wherein the zero-mode waveguide structure is a metal structure with nano holes on a dielectric substrate, and the nano-photon injection structure is a structure formed by dielectric materials with different refractive indexes. The zero mode waveguide structure or the nanophotonic ejection structure is a prior art, and the detailed structure thereof is not described herein.
The super-surface lens 1 is used for collecting fluorescence signals excited by the nano-structures 3, and the fluorescence signals with different wavelengths are focused to different focus positions of the same focal plane.
The transmission layer 5 is used for enabling the super surface lens 1 to have a certain distance with the detector 6, and ensuring that the fluorescence signal collected by the super surface lens 1 is focused on the detector 6. The transmission layer 5 may be silicon dioxide, hafnium dioxide or titanium dioxide, etc.
The detector 6 is positioned on the focal plane of the super surface lens 1 and is used for receiving fluorescence signals of various wavelengths focused by the super surface lens 1.
To ensure that different fluorescence signals are effectively distinguished by the detector 6, the distance between two adjacent focal points focused by the super-surface lens 1 is greater than 2 um.
The fluorescence signal collecting system unit provided by the invention can respond to fluorescence signals with different wavelengths through the super-surface lens 1, focus each fluorescence signal to different focus positions of the same focal plane, realize the function of the optical filter and cancel the use of the optical filter. And because the super-surface lens 1 has a simple structure, the manufacturing process is simpler than that of a Fresnel lens. Therefore, the super-surface lens provided by the invention is much lower than a traditional fluorescence signal collecting system combining a Fresnel lens and an optical filter in complexity, manufacturing process and cost.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
The above embodiments of the present invention should not be construed as limiting the scope of the present invention. Any other corresponding changes and modifications made according to the technical idea of the present invention should be included in the protection scope of the claims of the present invention.
Claims (9)
1. A super-surface lens is characterized by comprising a substrate and a medium layer prepared on the substrate, wherein a unit structure array is engraved on the medium layer and used for focusing incident light with different wavelengths in an incident light waveband at different focal positions on the same focal plane, and the distance between two adjacent focal points is larger than 2 microns.
2. The super surface lens of claim 1, wherein the numerical aperture of the super surface lens is greater than 0.4.
3. The super-surface lens according to claim 1, wherein the substrate is a silicon dioxide material, and the dielectric layer is a titanium dioxide material, a gallium nitride material, or a hafnium dioxide material.
4. The super surface lens of claim 1, wherein the shape of the unit structures in the unit structure array is rectangular, triangular, cylindrical, or elliptic cylindrical.
5. A fluorescence signal collection system comprising the super surface lens of any one of claims 1-4.
6. The fluorescence signal collection system of claim 5, further comprising a nanostructure, a waveguide structure, a transmission layer, and a detector, wherein said waveguide structure is fabricated on a substrate of said super-surface lens, said nanostructure is fabricated on said waveguide structure, said transmission layer is fabricated on an array of unit structures of said super-surface lens, and said detector is fabricated on said transmission layer.
7. The fluorescence signal collection system of claim 6, wherein the nanostructure is a zero mode waveguide structure or a nanophotonic ejection structure.
8. The fluorescence signal collection system of claim 6, wherein said detector is a CMOS or SCMOS detector.
9. The fluorescence signal collection system of claim 6, wherein the transmission layer is a silicon dioxide layer, a hafnium dioxide layer, or a titanium dioxide layer.
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| CN202011261855.1A CN112505009A (en) | 2020-11-12 | 2020-11-12 | Super surface lens and fluorescence signal collection system formed by same |
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Cited By (3)
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| CN114167604A (en) * | 2021-12-21 | 2022-03-11 | 福州大学 | Multi-sub-mirror array imaging element design method based on micro-size structure optimization |
| CN114280704A (en) * | 2021-12-28 | 2022-04-05 | 深圳迈塔兰斯科技有限公司 | Superlens array and wavefront detection system |
| CN116430678A (en) * | 2023-03-23 | 2023-07-14 | 华中科技大学 | Femtosecond laser direct writing system based on multifocal superlens |
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| CN116430678B (en) * | 2023-03-23 | 2024-02-06 | 华中科技大学 | Femtosecond laser direct writing system based on multifocal superlens |
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Application publication date: 20210316 |