CN115144377A

CN115144377A - Fluorescence collection device

Info

Publication number: CN115144377A
Application number: CN202110334498.5A
Authority: CN
Inventors: 不公告发明人
Original assignee: Shanghai Jinguan Technology Co ltd
Current assignee: Shanghai Jinguan Technology Co ltd
Priority date: 2021-03-29
Filing date: 2021-03-29
Publication date: 2022-10-04

Abstract

The utility model provides a fluorescence collection device, includes reaction layer, a plurality of collimation unit and signal collection layer, wherein, the reaction layer has the optical waveguide and is used for loading a plurality of biochemical reaction holes of determinand, the optical waveguide is used for producing the exciting light, makes determinand emission fluorescence signal, the collimation unit is right fluorescence signal collimates, the collimation unit with biochemical reaction hole one-to-one, and the biochemical reaction hole site is in the focus department of collimation unit, the signal collection layer is right fluorescence signal discerns the detection, when satisfying that equipment can be miniaturized, the position and the size of alignment unit place are controlled, have greatly improved optical detection's accuracy.

Description

Fluorescence collection device

Technical Field

The invention relates to the field of genetic engineering, in particular to a fluorescence collecting device.

Background

DNA sequencing (or DNA sequencing) refers to the analysis of the base sequence of a specific DNA fragment, i.e., the (G) arrangement of adenine (A), thymine (T), cytosine (C) and guanine.

Currently, techniques for sequencing mainly include a first-generation sequencing method using fluorescence sequencing of DNA, such as the dideoxynucleotide chain termination method (Sanger sequencing method), a second-generation sequencing method using cycle chip sequencing, and a third-generation sequencing method using single-molecule real-time sequencing technology of Pacific Biosciences, usa and Nanopore sequencing technology of Oxford nanopores Technologies, uk. Although the DNA fluorescent sequencing method has extremely high accuracy, the DNA fluorescent sequencing method in the prior art has the defects of difficult miniaturization of equipment, incapability of large-scale application and the like.

Therefore, a more perfect sequencing technology is still required to solve the disadvantages of the equipment such as no miniaturization, low precision and high cost.

Disclosure of Invention

The present invention provides a fluorescence collection device to solve some/all of the above technical problems.

In order to achieve the purpose, the invention provides the following technical scheme:

a fluorescence collecting device comprises a reaction layer, a plurality of collimation units and a signal collecting layer, and is provided with an optical waveguide and a plurality of biochemical reaction holes for loading objects to be tested, wherein the optical waveguide is used for generating exciting light to enable the objects to be tested to emit fluorescence signals, the collimation units correspond to the biochemical reaction holes one by one, and the biochemical reaction holes are positioned at the focuses of the collimation units. The reaction layer is provided with an optical waveguide and a plurality of biochemical reaction holes for loading objects to be detected, the optical waveguide is used for generating exciting light to enable the objects to be detected to emit fluorescent signals, the collimating unit collimates the fluorescent signals, and the signal collecting layer identifies and detects the fluorescent signals.

The collimation units are distributed in a periodic two-dimensional matrix, and aim to collimate the fluorescence in the reaction cavity to form parallel light which is collected and processed by the photoelectric detector.

The center distance of the adjacent collimating units is 500nm-10mm, preferably 10 μm-100 μm.

The collimating unit is located between the reaction layer and the signal collecting layer, the collimating unit is a lens array, and the lens array is provided with an anti-reflection film or an anti-reflection film so as to improve the collecting efficiency of fluorescence of a certain wave band or reduce the collecting efficiency of exciting light.

The lens array is one of a micro lens, a Fresnel lens and a super lens, the size of the lens is between 100nm and 100 mu m, and the micro lens is a spherical mirror or an aspherical mirror.

Compared with the prior art, the invention has the beneficial effects that:

1. the reaction layer of the fluorescence collecting device disclosed by the invention adopts the optical waveguide to enable the substance to be detected in the biochemical reaction hole to generate the fluorescence signal, and the collimation unit is used for collimating the fluorescence signal, so that the signal collecting layer is convenient to collect the fluorescence signal and carry out identification and detection, the position and the size of the collimation unit are controlled while the miniaturization of equipment is met, and the accuracy of optical detection is greatly improved.

2. In the fluorescence collecting device of the present invention, the collimating unit may be one of a micro lens, a fresnel lens, and a super lens, and the collimating unit further includes an antireflection film or a high reflection film to improve the collection efficiency of fluorescence of a specific wavelength or reduce the collection efficiency of excitation light, and the collimating unit may be replaced or changed as required, thereby more effectively improving the detection accuracy.

Drawings

In order to more clearly illustrate the technical solutions in the specific embodiments of the present invention, the drawings required to be used in the description of the embodiments will be briefly introduced below, and it is apparent that the drawings described below are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings may be obtained based on these drawings without inventive efforts.

FIG. 1 is a schematic view of a fluorescence collection device according to the present invention;

FIG. 2 is a schematic diagram of the working principle of a microlens;

FIG. 3 is a schematic view of the present invention with microlenses assembled;

FIG. 4 is a schematic view of the arrangement of a microlens array;

FIG. 5 is a schematic diagram of the working principle of the Fresnel lens;

FIG. 6 is a schematic view of the present invention with a Fresnel lens assembled;

FIG. 7 is a schematic layout diagram of a Fresnel lens array;

FIG. 8 is a two-dimensional top view of a superlens array;

FIG. 9 is a schematic diagram of the present invention assembled with a superlens array.

Reference numerals are as follows: the light source comprises a reaction layer 1, a biochemical reaction hole 11, a wrapping layer 12, an optical waveguide 13, a collimating unit 2, a lens structure 21, a fluorescent signal 22, a signal collecting layer 3, a light processing material layer 31, a pixel element 32, a light blocking component 33, a micro lens 4, a Fresnel lens 5 and a super lens 6.

Detailed Description

The technical solutions in the specific embodiments of the present invention will be clearly and completely described below, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example one

As shown in fig. 1, the present invention provides a fluorescence collecting device, which includes a reaction layer 1, a plurality of collimating units 2, and a signal collecting layer 3, wherein the reaction layer 1 includes a biochemical reaction hole 11, an optical waveguide 13, and a wrapping layer 12, and the biochemical reaction hole 11 is one of a square, a circle, a semicircle, an ellipse, or a polygon; the cladding layer 12 is one of an optically transparent polymer material or silicon dioxide, and the optical waveguide 13 is made of an optically transparent material with a high refractive index, such as silicon nitride.

The collimating unit 2 is a lens structure 21 for collimating the fluorescent signal 22 and transmitting the fluorescent signal 22 to the signal collection layer 3.

The signal collection layer 3 includes a light processing material layer 31, a light blocking component 33, and a pixel element 32, wherein the light processing material layer 31 is a filter material layer or a light splitting material layer, and when the filter material layer is used, the filter material layer can be filter layers with different optical properties, and the filter wavelength bands of the filter layers can be different, so as to detect fluorescent signals of different bases; when the light splitting material layer is used, the light splitting material layer can be used for separating optical signals of different emission wave bands, the light splitting material layer can be a grating or a prism, and fluorescent signals in the biochemical reaction holes 11 are spread on the inner space of the light splitting material layer and projected on different pixel elements 32; and the detection of signals of different wave bands is realized. The number of the pixel elements 32 can be one or several, and the size of the pixel elements 32 is 2 μm-100 μm.

When the lens structure 21 is a microlens 4, as shown in fig. 2 to 4, it is arranged in two-dimensional periodic arrangement, corresponding to the upper biochemical reaction hole 11. The purpose of the micro-lens 4 array is to collimate the fluorescent signal 22 in the biochemical reaction well 11, and the fluorescent signal 22 emitted by the biochemical reaction is collimated by the micro-lens 4 to form parallel light which is collected and processed by the signal collection layer 3.

In this embodiment, the pixel element 32 in the signal collection layer may be a CMOS Image Sensor (CIS), a PhotoMultiplier Tube (PMT), a Single Photon Avalanche Diode (SPAD), a Charge-Coupled Device (CCD), a Silicon PhotoMultiplier (SiPM), or the like, but is not limited thereto.

The size range of the micro lenses 4 is 100nm-100 μm, the center distance of the micro lenses depends on the center distance of the biochemical reaction holes 11, the larger the theoretical spatial distribution density is, the better the theoretical spatial distribution density is, the high-throughput detection is favorably realized, meanwhile, the processing technology limits are met, the crosstalk among different channels is avoided, the center distance between the adjacent micro lenses 4 is 500nm-10mm, and the preferable range is 10 μm-100 μm. The biochemical reaction hole 11 is positioned at the focus of the micro lens 4.

Optionally, the micro-lens 4 is coated with an Anti-reflection coating (Anti-reflection coating) to improve the collection efficiency of fluorescence in a certain wavelength band, and optionally coated with a High-reflection coating (High-reflection coating) to reduce the collection efficiency of excitation light. The processing of the microlens 4 can be realized by Reactive ion etching (Reactive ion etching) and photolithography (photoresist).

Fig. 4 is an optical path diagram of a fluorescence signal 22 when the lens structure 21 of the collimating unit 2 of the present invention is a microlens 4.

The micro lens 4 can be a spherical lens, and in order to improve the detection performance of the system, the micro lens 4 can also be an aspheric lens; the microlens 4 material may be a polymer material, silicon dioxide, or the like.

Example two

As shown in fig. 5 to 7, when the fresnel lens 5 is formed by the lens structure 21, the thickness and weight of the lens structure 21 and the overall thickness of the system can be reduced. In this embodiment, the fresnel lenses 5 are periodically arranged in two dimensions and form a corresponding relationship with the biochemical reaction holes 11. The purpose of the fresnel lens 5 is to collimate the fluorescent signal 22 in the biochemical reaction hole 11, and the fluorescence emitted by the biochemical reaction is collimated by the lens to form parallel light, which is collected and processed by the signal collection layer 3.

The size of the Fresnel lens 5 is 100nm-100 μm, the larger the theoretical spatial distribution density is, the better the Fresnel lens is, the higher the theoretical spatial distribution density is, the higher the detection is, the higher the throughput is, while being limited by the processing technology and avoiding cross-talk between different channels, the lens center distance is 500nm-10mm, preferably 10 μm-100 μm. The biochemical reaction hole 11 is positioned at the focus of the Fresnel lens 5. The fresnel lens 5 may be optionally coated with Anti-reflection coatings (Anti-reflection coatings) to improve the collection efficiency of fluorescence in a certain wavelength band, and may be optionally coated with a High-reflection coating (High-reflection coating) to reduce the collection efficiency of excitation light.

Fig. 7 is a schematic diagram of a lens array when the lens structure 21 of the collimating unit 2 of the present invention is a fresnel lens 5. The fresnel lens 5 can be manufactured by injection molding, reactive ion Etching (Reactive ion Etching), photolithography (photonic), and the like.

EXAMPLE III

To further reduce the thickness, weight, and overall thickness of the system of lens structures 21, as shown in fig. 8 and 9, a super-structured planar lens, such as an array of superlenses 6 using metamaterials, may be employed. The superlens 6 has an ultra-thin and flat structure, the surface is an array of waveguides like micro-struts arranged in a specific pattern. These pillars are various nano-elements, with a length of around 600nm, and may be made of materials such as TiO ₂ And the like. Since it is flat and ultraThe thin feature, all wavelengths of light pass almost simultaneously, the superlens 6 has the advantage of achromatic and tunable dispersion. The superlens 6 array arranged periodically in two dimensions and the biochemical reaction holes 11 above form a one-to-one correspondence relationship.

The super lens 6 collimates the fluorescence signal 22 in the biochemical reaction hole 11, and the biochemical reaction hole 11 is located at the focus of the super lens 6. In the embodiment, the size of the superlens 6 is 100nm-100 μm, the larger the theoretical spatial distribution density is, the better the superlens is, the high-flux detection is realized, while being limited by the processing technology and avoiding crosstalk between different channels, the lens center distance is 500nm-10mm, preferably 10 μm-100 μm. The super lens 6 can be produced in the existing CMOS semiconductor foundry in large scale, and is beneficial to realizing the large-scale wafer-level integration of the optical system.

The fluorescence collecting device provided by the present invention is described in detail above, and the structure and the operation principle of the present invention are explained by applying specific examples, and the above description of the embodiments is only used to help understanding the method and the core idea of the present invention. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, it is possible to make various improvements and modifications to the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.

Claims

1. A fluorescence collection device, comprising:

the reaction layer is provided with an optical waveguide and a plurality of biochemical reaction holes for loading an object to be detected, wherein the optical waveguide is used for generating exciting light so that the object to be detected emits a fluorescent signal;

the collimation units are arranged on the light path of the fluorescence signal and collimate the fluorescence signal;

the signal collection layer is positioned on the light path of the collimated fluorescent signal and is used for identifying and detecting the fluorescent signal;

wherein, the plurality of collimation units correspond to the plurality of biochemical reaction holes one by one;

the biochemical reaction hole is positioned at the focus of the collimation unit.

2. The fluorescence collection device of claim 1, wherein the plurality of collimating elements form a periodic two-dimensional matrix.

3. The fluorescence collection device of claim 2, wherein the centers of adjacent collimating units are spaced between 500nm-10mm apart.

4. The fluorescence collection device of claim 1, wherein the collimating unit is a lens structure.

5. The fluorescence collection device of claim 4, wherein the lens structure is disposed between the reaction layer and the signal collection layer.

6. The fluorescence collection device of claim 4, wherein said lens structure has an anti-reflection film or a highly reflective film.

7. The fluorescence collection device of claim 4, wherein the lens structure is between 100nm-100 μm in size.

8. The fluorescence collection device of claim 4, wherein said lens structure is one of a microlens, a Fresnel lens, and a superlens.

9. The fluorescence collection device of claim 8, wherein the microlens is a spherical mirror or an aspherical mirror.