CN117554349B - Nano integrated optical chip for single molecule sensing and fluorescence detection method - Google Patents
Nano integrated optical chip for single molecule sensing and fluorescence detection method Download PDFInfo
- Publication number
- CN117554349B CN117554349B CN202410039508.6A CN202410039508A CN117554349B CN 117554349 B CN117554349 B CN 117554349B CN 202410039508 A CN202410039508 A CN 202410039508A CN 117554349 B CN117554349 B CN 117554349B
- Authority
- CN
- China
- Prior art keywords
- excitation
- light
- layer
- micro
- waveguide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000003287 optical effect Effects 0.000 title claims abstract description 66
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000001917 fluorescence detection Methods 0.000 title claims abstract description 15
- 230000005284 excitation Effects 0.000 claims abstract description 260
- 230000005540 biological transmission Effects 0.000 claims abstract description 79
- 238000001514 detection method Methods 0.000 claims abstract description 71
- 239000010410 layer Substances 0.000 claims description 129
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 26
- 239000000463 material Substances 0.000 claims description 21
- 238000010168 coupling process Methods 0.000 claims description 19
- 230000008878 coupling Effects 0.000 claims description 18
- 238000005859 coupling reaction Methods 0.000 claims description 18
- 238000005253 cladding Methods 0.000 claims description 15
- 239000012792 core layer Substances 0.000 claims description 15
- 239000000377 silicon dioxide Substances 0.000 claims description 13
- 229910021417 amorphous silicon Inorganic materials 0.000 claims description 11
- HBMJWWWQQXIZIP-UHFFFAOYSA-N silicon carbide Chemical compound [Si+]#[C-] HBMJWWWQQXIZIP-UHFFFAOYSA-N 0.000 claims description 11
- 235000012239 silicon dioxide Nutrition 0.000 claims description 9
- 239000007850 fluorescent dye Substances 0.000 claims description 8
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 6
- 229910052710 silicon Inorganic materials 0.000 claims description 6
- 239000010703 silicon Substances 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 239000004038 photonic crystal Substances 0.000 claims description 5
- 238000005286 illumination Methods 0.000 abstract description 33
- 210000004027 cell Anatomy 0.000 description 14
- 230000002238 attenuated effect Effects 0.000 description 6
- 210000000170 cell membrane Anatomy 0.000 description 6
- 238000010586 diagram Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 230000001965 increasing effect Effects 0.000 description 5
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000002457 bidirectional effect Effects 0.000 description 4
- 238000004557 single molecule detection Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000010494 dissociation reaction Methods 0.000 description 3
- 230000005593 dissociations Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 238000009792 diffusion process Methods 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 230000035699 permeability Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 238000000411 transmission spectrum Methods 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000001312 dry etching Methods 0.000 description 1
- 238000005530 etching Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 238000000059 patterning Methods 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 238000000623 plasma-assisted chemical vapour deposition Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Classifications
-
- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
-
- 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
- G01N21/03—Cuvette constructions
- G01N21/0303—Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment
-
- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
-
- 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
- G01N21/03—Cuvette constructions
- G01N2021/0346—Capillary cells; Microcells
-
- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6482—Sample cells, cuvettes
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention relates to the technical field of fluorescence detection, and discloses a nano integrated optical chip for single-molecule sensing and a fluorescence detection method, wherein the chip comprises the following components: a fluorescent transmission layer with a detection area on the surface; at least two groups of light input units, the input ends of the light input units are connected with an external laser light source, and the output ends of the light input units are used for introducing excitation light into the detection area; the at least one group of excitation waveguide units are arranged in the detection area and comprise an excitation waveguide body and a plurality of micro-ring resonant cavities, and two ends of the excitation waveguide body are respectively connected with the output ends of the two groups of light input units; the micro-ring resonant cavities are arranged at intervals at the side of the excitation waveguide body; the micro-ring resonant cavity is annular, the distance between the center of the micro-ring resonant cavity and the center of the excitation waveguide body is smaller than half of the wavelength of the excitation light, and the circumference of the micro-ring resonant cavity is integral multiple of the wavelength of the excitation light. Excitation light resonates and enhances in the micro-ring resonant cavity, so that the illumination area and the intensity consistency of the excitation light are improved, and the detection efficiency and the detection accuracy are improved.
Description
Technical Field
The invention relates to the technical field of fluorescence detection, in particular to a nano integrated optical chip for single-molecule sensing and a fluorescence detection method.
Background
Unlike conventional biochemical sensing methods based on average molecular population, single-molecule sensing technology can reflect dynamic heterogeneity of single molecules, and thus, detection technology related to single molecules is gaining more and more attention. The current single-molecule detection technology is mostly based on microscopic imaging, relies on complex, huge and expensive microscopic optical path systems, and is used for integrating optical path modules and functions of laser excitation, guidance, focusing, beam splitting, fluorescence sensing and the like in a very small space, so that the current single-molecule detection technology can replace the huge microscopic optical path systems, and as tens or even hundreds of single-molecule sensing chips with the functions can be manufactured on one wafer, the cost is greatly reduced, and the commercialization of the single-molecule sensing technology is possible.
In order to improve the accuracy of single-molecule detection, it is necessary to perform multi-site parallel detection on a single chip, and the number of detection sites of the single chip is often set to tens of thousands to millions or even more. The same light source or light conducting element is usually used to provide excitation light to the detection site, for example, an optical waveguide element using the principle of total internal reflection effect, and an evanescent field is formed around the surface of the optical waveguide element, thereby forming an illumination region. And adding a substance to be detected to a detection site, wherein fluorescent substances contained in the substance to be detected are excited to generate fluorescence emission light, and the fluorescence emission light is sensed by a sensing structure to finish optical detection of the substance to be detected.
However, with the long-distance excitation optical waveguide element, the single-point illumination area formed around the optical waveguide surface is small, the excitation light of the same optical waveguide element is attenuated with the increase of the transmission distance, and the problem of inconsistent excitation light intensity exists among different branch optical waveguides. Because of the difference in coupling efficiency caused by the distance between different optical waveguides and the laser light source and the structure obtained by actual processing, the excitation light illumination of different areas of the chip is uneven, and the emitted light loss is different, the signals emitted by all detection sites are finally not at the same intensity level or even distorted, so that interference is caused, and the accuracy of multi-point parallel detection on the same chip is difficult to ensure.
Disclosure of Invention
In view of the above, the invention provides a nano integrated optical chip for single-molecule sensing and a fluorescence detection method, which are used for solving the problems that in the existing single-molecule sensing detection, the excitation light of each detection site on the chip has small illumination area and inconsistent intensity, so that the emitted signal intensities are different, and the detection accuracy is difficult to guarantee.
In a first aspect, the present invention provides a nano-integrated optical chip for single molecule sensing, comprising: a fluorescent transmission layer, at least two groups of light input units and at least one group of excitation waveguide units. The surface of the fluorescent transmission layer is provided with a detection area; the input end of any light input unit is connected with an external laser light source, and the output end of any light input unit stretches into the detection area so as to introduce excitation light into the detection area; at least one group of excitation waveguide units are arranged on the fluorescent transmission layer and positioned in the detection area; the excitation waveguide unit comprises an excitation waveguide body and a plurality of micro-ring resonant cavities, and two ends of the excitation waveguide body are respectively connected with the output ends of the two groups of light input units so as to bidirectionally input excitation light; the micro-ring resonant cavities are arranged at intervals at the side of the excitation waveguide body along the direction parallel to the excitation waveguide body; the micro-ring resonant cavities are in a circular ring shape, the distance between the center of any micro-ring resonant cavity and the center of the excitation waveguide body is smaller than half of the wavelength of the excitation light along the direction perpendicular to the excitation waveguide body, and the circumference of each micro-ring resonant cavity is set to be integral multiple of the wavelength of the excitation light.
The beneficial effects are that: according to the invention, the excitation light of the external excitation light source is input and output in the excitation waveguide body in a bidirectional manner, so that the attenuation of the excitation light unidirectionally transmitted in the same excitation waveguide body along with the increase of the transmission distance is avoided, the intensity of the excitation light in the same excitation waveguide body is consistent, and the intensity of the excitation light transmitted subsequently is ensured not to be attenuated. And a plurality of micro-ring resonant cavities are arranged around each excitation waveguide body, and the distance L between the center of the excitation waveguide body and the center of the micro-ring resonant cavity is smaller than half of the wavelength of excitation light so as to ensure that the excitation light is coupled between the excitation waveguide body and the micro-ring resonant cavities, and excitation light of the excitation waveguide body is coupled into the micro-ring resonant cavities. The circumference of the center of the micro-ring resonant cavity is set to be an integral multiple of the wavelength of excitation light, so that excitation light is subjected to resonance enhancement in the micro-ring resonant cavity; the micro-ring resonant cavity extends the illumination area of excitation light at a certain point on the linear excitation waveguide body, one point becomes a ring, and the illumination intensity of the same ring is absolutely consistent, thereby being beneficial to improving the detection accuracy and efficiency.
In an alternative embodiment, the optical input unit comprises a coupling grating and a transmission waveguide; one end of the coupling grating is connected with an external laser light source, the other end of the coupling grating is connected with one end of the transmission waveguide, and the other end of the transmission waveguide is connected with the excitation waveguide body.
In the invention, the coupling grating is adopted, so that the excitation light from an external laser emitter can smoothly enter the transmission waveguide on the chip, and the transmission waveguide stably transmits the excitation light into the excitation waveguide body.
In an alternative embodiment, the transmission waveguide, the excitation waveguide body and the micro-ring resonator are made of the same material, and each of the transmission waveguide, the excitation waveguide body and the micro-ring resonator comprises a core layer and a cladding layer, wherein the refractive index of the core layer is larger than that of the cladding layer.
In the invention, the materials of the transmission waveguide, the excitation waveguide body and the micro-ring resonant cavity are the same, so as to realize consistent light conduction.
In an alternative embodiment, the material of the core layer is amorphous silicon carbide, and the refractive index of the core layer ranges from 2.3 to 3.0; the cladding is made of silicon dioxide or aqueous solution; the outer surface of the micro-ring resonant cavity is provided with an excitation area, and the thickness of the excitation area is 50nm.
According to the invention, the high refractive index of the amorphous silicon carbide reduces the penetration thickness of an evanescent field around the micro-ring resonant cavity, reduces the volume of an excitation area, reduces the interference of free molecules, and is more beneficial to single-molecule sensing detection of target detection molecules. On the premise of exciting the illumination area of the waveguide body, the micro-ring resonant cavity extends the illumination area of excitation light, so that the detection efficiency and accuracy are improved.
In an alternative embodiment, multiple sets of excitation waveguide units are arranged parallel to each other; the transmission waveguides in the two groups of optical input units have a multi-stage Y-shaped bifurcated structure.
In the invention, the input end of the transmission waveguide is a single interface and is connected with the coupling grating, the output end of the transmission waveguide is a branched multi-interface and is further connected with a plurality of linear excitation waveguides, the multi-interface output end of one transmission waveguide is connected with the upper end of the excitation waveguide, and the multi-interface output end of the other transmission waveguide is connected with the lower end of the excitation waveguide, so that the uniform excitation light illumination area on a plurality of detection sites is provided.
In an alternative embodiment, the method further comprises: and the image sensing unit is arranged on the surface of one side, away from the excitation waveguide unit, of the fluorescence transmission layer.
In the invention, the image sensing unit is correspondingly arranged with the excitation waveguide unit above the fluorescence transmission layer and is used for receiving the kinetic information of the single molecule to be detected in the sample to be detected in the detection area.
In an alternative embodiment, the fluorescent light-transmitting layer comprises a wavelength-selective layer; the excitation waveguide unit is disposed on the wavelength selective layer.
In the invention, the wavelength selective layer has selective permeability to fluorescence emission light of a specific wave band, selectively transmits single-molecule fluorescence, blocks stray light such as excitation light and the like, and suppresses noise, thereby avoiding signal interference caused by the transmission of the excitation light, further enhancing the suppression of the excitation light, and avoiding loss of the fluorescence emission light.
In an alternative embodiment, the material of the wavelength selective layer is isotropic amorphous silicon carbide.
In the invention, the amorphous silicon carbide can block a part of excitation light and allow light in a wave band where fluorescence emission light is located to pass, thereby effectively preventing transmission loss of the fluorescence emission light.
In an alternative embodiment, the material of the wavelength selective layer is a photonic crystal material consisting of silicon layers and silicon dioxide layers alternately, the thickness of the silicon layers and the silicon dioxide layers being in the range of 100nm to 400nm.
In the invention, the photon crystal material can also realize blocking of the excitation light and transmitting of the fluorescence emission light, and the material is simple and stable.
In an alternative embodiment, the fluorescent light-transmitting layer further comprises a fluorescent guiding layer; the fluorescence guiding layer is arranged between the wavelength selection layer and the excitation waveguide unit, the excitation waveguide unit is arranged on the fluorescence guiding layer, and the detection area is arranged on the surface of the fluorescence guiding layer.
In the invention, the fluorescence guiding layer can enable fluorescence emitted light to be transmitted to the wavelength selection layer as far as possible so as to reach the image sensing unit.
In an alternative embodiment, the thickness of the fluorescent guiding layer ranges from 1um to 5um.
In the invention, the thickness of the fluorescent guiding layer is used for controlling the distance between the plane where the excitation waveguide unit is located and the image sensing unit, the too small distance can cause signal interference caused by the fact that the light evanescent field around the excitation waveguide body and the micro-ring resonant cavity enters the image sensor, and the too large distance can cause larger loss when the fluorescent emission light around the excitation waveguide body and the micro-ring resonant cavity reaches the image sensor.
In a second aspect, the present invention provides a fluorescence detection method applied to the above nano integrated optical chip for single molecule sensing, including:
adding a sample to be detected to a detection area of the fluorescent transmission layer, wherein the sample to be detected comprises single molecules to be detected;
adding a binding molecule containing fluorescent labels into a detection area of the fluorescent transmission layer, wherein the binding molecule is combined with a single molecule to be detected;
the nano integrated optical chip for single molecule sensing is controlled to emit excitation light, so that the fluorescent label is excited to emit fluorescence emission light, and the fluorescence emission light is emitted from the fluorescence transmission layer.
The beneficial effects are that: and (3) judging the binding or dissociation event between the single molecule to be detected and the binding molecule by capturing a fluorescent signal emitted when the binding molecule with the fluorescent label is excited, so as to obtain the kinetic information of the single molecule interaction. In the process, the excitation light of the external excitation light source is input and output in the excitation waveguide body in a bidirectional manner, so that the intensity of the excitation light on the same excitation waveguide body is consistent, and the intensity of the excitation light transmitted subsequently is ensured not to be attenuated; meanwhile, the micro-ring resonant cavity extends the excitation light illumination area of a certain point on the linear excitation waveguide body, one point becomes a ring, the illumination intensity of the same ring is absolutely consistent, and the accuracy and the efficiency of single-molecule sensing detection are improved.
In an alternative embodiment, the sample to be measured covers at least one micro-ring resonator.
According to the invention, cells in a sample to be detected cross one or two or more micro-ring resonant cavities, the circular light field on the space of the micro-ring resonant cavities increases the area of single cells illuminated by excitation light from the same excitation waveguide body, the intensity consistency of excitation light in local areas of the cells and the total illumination area are increased, the detection efficiency is further increased, and compared with a linear excitation waveguide, the micro-ring resonant cavities have larger contact area corresponding to cell membranes, so that the sensing area is larger, and the efficiency of carrying out large-area single-molecule parallel detection on the cell surfaces is high.
In an alternative embodiment, the method further comprises: an image sensing unit is arranged on a surface of the fluorescent light transmission layer, which surface is away from the excitation waveguide unit, and the image sensing unit is suitable for receiving fluorescent light emission.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of a nano-integrated optical chip for single molecule sensing according to an embodiment of the present invention;
FIG. 2 is an enlarged view of the invention shown in section A of FIG. 1;
FIG. 3 is a top view of an excitation waveguide unit in a nano-integrated optical chip for single molecule sensing according to an embodiment of the present invention;
FIG. 4 is a schematic diagram of a simulation of the optical field of a nano-integrated optical chip excitation waveguide unit for single molecule sensing according to an embodiment of the present invention;
FIG. 5 is a partial cross-sectional view of a nano-integrated optical chip for single molecule sensing according to an embodiment of the present invention;
FIG. 6 is a schematic diagram of an excitation region in a nano-integrated optical chip for single molecule sensing according to an embodiment of the present invention;
FIG. 7 is a transmission spectrum of a wavelength selective layer in a nano-integrated optical chip for single molecule sensing according to an embodiment of the present invention;
FIG. 8 is a cross-sectional view of a single molecule to be tested in combination with a binding molecule of a sample to be tested in an embodiment of the present invention;
FIG. 9 is a schematic diagram of a sample to be tested and an excitation waveguide unit according to an embodiment of the present invention.
Reference numerals illustrate:
1. a fluorescent light transmitting layer; 11. a wavelength selective layer; 12. a fluorescent guiding layer;
2. a light input unit; 21. a coupling grating; 22. a transmission waveguide;
3. exciting the waveguide unit; 31. exciting the waveguide body; 32. a micro-ring resonator; 3201. resonating incident light; 3202. resonating light; 3203. resonating outgoing light;
4. an excitation region;
5. an image sensing unit;
100. a sample to be tested; 101. a single molecule to be measured; 200. a binding molecule; 300. and (5) an aqueous solution to be measured.
Detailed Description
The invention is described in further detail below with reference to the drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting thereof. It should be further noted that, for convenience of description, only some, but not all of the structures related to the present invention are shown in the drawings. In the following description, descriptions of well-known structures and techniques are omitted so as not to unnecessarily obscure the concepts of the present invention. Various structural schematic diagrams according to embodiments of the present invention are shown in the accompanying drawings. The figures are not drawn to scale, wherein certain details are exaggerated for clarity of presentation and may have been omitted. The shapes of the various regions, layers and relative sizes, positional relationships between them shown in the drawings are merely exemplary, may in practice deviate due to manufacturing tolerances or technical limitations, and one skilled in the art may additionally design regions/layers having different shapes, sizes, relative positions as actually required. In the context of the present invention, when a layer/element is referred to as being "on" another layer/element, it can be directly on the other layer/element or intervening layers/elements may be present therebetween. In addition, if one layer/element is located "on" another layer/element in one orientation, that layer/element may be located "under" the other layer/element when the orientation is turned.
In the related art, the integrated optical chip for single molecule sensing can integrate optical path modules and functions such as laser excitation, guiding, focusing, beam splitting, fluorescence sensing and the like in a very small space, so that the dynamic heterogeneity of single molecules is reflected. In order to improve the accuracy of single molecule detection, multi-site parallel detection is usually performed on a single chip, and the same light source or light conduction element is used to provide excitation light to the detection site, for example, an optical waveguide element using the principle of total internal reflection effect, so that an evanescent field is formed around the surface of the optical waveguide element, and an illumination area is formed.
However, when the excitation optical waveguide element with a long distance is used, the illumination area of the single point of the excitation light formed around the surface of the optical waveguide is small, the excitation light of the same optical waveguide element is attenuated with the increase of the transmission distance, and the problem of inconsistent intensity of the excitation light is more existed between different branch optical waveguides. Because of the difference in coupling efficiency caused by the distance between different optical waveguides and the laser light source and the structure obtained by actual processing, the excitation light illumination of different areas of the chip is uneven, and the emitted light loss is different, the signals emitted by all detection sites are finally not at the same intensity level or even distorted, so that interference is caused, and the accuracy of multi-point parallel detection on the same chip is difficult to ensure.
The embodiment of the invention provides a nano integrated optical chip for single-molecule sensing, which is used for the bidirectional transmission of excitation light in an excitation waveguide body 31, solves the problem that the excitation light is attenuated along with the increase of transmission distance during unidirectional transmission, and a plurality of micro-ring resonant cavities 32 arranged near the same excitation waveguide body 31 extend the illumination area of the excitation light in the excitation waveguide body 31, thereby achieving the effects of increasing the single-point illumination area of the excitation light and ensuring the uniform illumination intensity of the excitation light.
As shown in fig. 1 to 7, a nano-integrated optical chip for single molecule sensing according to an embodiment of the present invention includes:
a fluorescent transmission layer 1, at least two groups of light input units 2 and at least one group of excitation waveguide units 3;
the surface of the fluorescent transmission layer 1 is provided with a detection area; the input end of the light input unit 2 is connected with an external laser light source, and the output end of the light input unit extends into the detection area so as to introduce excitation light into the detection area; the excitation waveguide unit 3 is arranged on the fluorescent transmission layer 1 and is positioned in the detection area; the excitation waveguide unit 3 comprises an excitation waveguide body 31 and a plurality of micro-ring resonant cavities 32, and two ends of the excitation waveguide body 31 are respectively connected with the output ends of the two groups of optical input units 2 so as to bidirectionally input excitation light; the micro-ring resonators 32 are arranged at intervals beside the excitation waveguide body 31 along the direction parallel to the excitation waveguide body 31; the micro-ring resonators 32 are ring-shaped, the distance between the center of any micro-ring resonator 32 and the center of the excitation waveguide body 31 is less than half of the wavelength of the excitation light along the direction perpendicular to the excitation waveguide body 31, and the circumference of the micro-ring resonator 32 is set to be an integer multiple of the wavelength of the excitation light.
In the nano integrated optical chip for single molecule sensing, a fluorescent transmission layer 1 forms a chip substrate, and a light input unit 2 and an excitation waveguide unit 3 are integrated on one side surface of the fluorescent transmission layer 1, wherein a region on the fluorescent transmission layer 1 for adding a sample 100 to be detected for fluorescent sensing detection is called a detection region. The excitation waveguide units 3 are arranged in the detection area, the excitation waveguide body 31 is a linear waveguide, two ends of the excitation waveguide body are respectively connected with the output ends of the two groups of light input units 2, excitation light of an external excitation light source is input and output in the excitation waveguide body 31 in a bidirectional mode, further attenuation caused by the fact that the excitation light unidirectionally transmitted in the same excitation waveguide body 31 is increased along with the increase of the transmission distance is avoided, the intensity of the excitation light in the same excitation waveguide body 31 is consistent, and the intensity of the excitation light subsequently transmitted is guaranteed not to be attenuated.
On this basis, a plurality of micro-ring resonators 32 are arranged around each excitation waveguide body 31, as shown in fig. 5, the distance L between the center of the excitation waveguide body 31 and the center of the micro-ring resonators 32 is smaller than half the wavelength of the excitation light, so as to ensure that the excitation light is coupled between the excitation waveguide body 31 and the micro-ring resonators 32, and the excitation light of the excitation waveguide body 31 is coupled into the micro-ring resonators 32. The circumference of the center of the micro-ring resonant cavity 32 is set to be an integral multiple of the wavelength of the excitation light, so that the excitation light is subjected to resonance enhancement in the micro-ring resonant cavity 32; the micro-ring resonator 32 extends the illumination area of excitation light at a point on the linear excitation waveguide body 31, one point becomes a ring, and the illumination intensity of the same ring is absolutely uniform. The illumination area of the present embodiment is increased and the illumination intensity is kept uniform, as compared with the single excitation waveguide body 31, contributing to the improvement of detection accuracy and efficiency.
Specifically, as shown in fig. 3, the coupling process of the excitation light between the excitation waveguide body 31 and the micro-ring resonator 32 is: the excitation light transmitted in the excitation waveguide body 31 forms an evanescent field on the peripheral surface of the excitation waveguide body 31, and a portion of light in the evanescent field, which is close to the micro-ring resonator 32, is coupled into the micro-ring resonator 32 in the form of resonance incident light 3201, is transmitted in the micro-ring resonator 32 in a total reflection manner in the form of resonance light 3202, is emitted into the excitation waveguide body 31 in the form of resonance emergent light 3203, and is transmitted back in the excitation waveguide body 31 in the form of excitation light. The above-described resonance light 3202 is substantially the same as the excitation light, and the excitation light in the ring structure of the micro-ring resonator 32 is referred to as resonance light 3202, which will be collectively referred to as excitation light in the following description. Simulation calculation results of coupling of excitation light between the excitation waveguide body 31 and the micro-ring resonator 32 are shown in fig. 4. The excitation light in the excitation waveguide body 31 is bidirectionally input along the linear excitation waveguide body 31 and bidirectionally output, so that the attenuation of the excitation light to zero due to the coupling with the micro-ring resonant cavity 32 is avoided, and the intensity of the excitation light transmission is ensured.
As shown in fig. 1, the optical input unit 2 includes a coupling grating 21 and a transmission waveguide 22, one end of the coupling grating 21 is connected to an external laser light source, the other end of the coupling grating 21 is connected to one end of the transmission waveguide 22, and the other end of the transmission waveguide 22 is connected to an excitation waveguide body 31. The two sets of light input units 2 have two sets of excitation light output ends, which are respectively connected with two ends of the linear excitation waveguide body 31. The external laser light source is usually a laser emitter, which is usually an optical fiber interface, the generated excitation light cannot be directly coupled into the optical waveguide structure on the chip, and the coupling grating 21 is adopted to enable the excitation light from the external laser emitter to smoothly enter the transmission waveguide 22 on the chip, so that the transmission waveguide 22 stably transmits the excitation light into the excitation waveguide body 31.
In one embodiment, the materials of the transmission waveguide 22, excitation waveguide body 31, and micro-ring resonator 32 are the same to achieve uniform light conduction.
Specifically, the transmission waveguide 22, the excitation waveguide body 31, and the micro-ring resonator 32 each include a core layer and a cladding layer, and the refractive index of the core layer is greater than that of the cladding layer. The excitation light is totally reflected between the core layer and the cladding layer, and the excitation light can only be transmitted within a very small distance in the cladding layer near the interface of the two layers, and the transmitted light is called an evanescent field. The illumination area of the excitation light, i.e. the illumination area of the evanescent field generated during the transmission of the excitation light, can be understood as the planar area of the evanescent field. Taking the micro-ring resonant cavity 32 as an example, due to the refractive index difference between the core layer and the cladding layer of the micro-ring resonant cavity 32, the excitation light generated by resonance, namely, the resonant light 3202, is transmitted in the micro-ring resonant cavity 32 in a total internal reflection manner, an evanescent field is formed around and on the surface of the micro-ring resonant cavity 32, the illumination area of the excitation light in the micro-ring resonant cavity 32 is the area of the micro-ring resonant cavity 32 forming the evanescent field on the plane parallel to the surface of the chip, and similarly, the illumination area of the excitation light in the excitation waveguide body 31 is the area of the excitation waveguide body 31 forming the evanescent field on the plane parallel to the surface of the chip.
As a preferred embodiment, the core layer material is amorphous silicon carbide, and the refractive index of the core layer ranges from 2.3 to 3.0; the cladding layer is made of silicon dioxide or an aqueous solution, so that the outer surface of the micro-ring resonator 32 forms an excitation area 4, that is, a space of an evanescent field around and on the micro-ring resonator 32 in the thickness direction, and the thickness of the excitation area 4 is 50nm, as shown in fig. 6. The thickness of the evanescent field depends on the refractive index difference between the core and the cladding, the larger the difference the smaller the evanescent field. The high refractive index of amorphous silicon carbide reduces the penetration thickness of the evanescent field formed by the micro-ring resonator 32, i.e. reduces the volume of the excitation region 4. The diffusion speed of the free molecules is constant, and the smaller the volume of the excitation region 4 formed by the evanescent field is, the shorter the residence time of the free molecules in the excitation region 4 is, and the time of the free molecules in the excitation region 4 is in the order of microseconds. The target detection molecules are fixed in the chip detection area and are not influenced by the excitation area 4, and the residence time is in the millisecond level, so that the thickness of the excitation area 4 is 50nm, the excitation volume is small, the interference of free molecules is reduced, and the single-molecule sensing detection of the target detection molecules is facilitated. Meanwhile, the micro-ring resonant cavity 32 extends the illumination area of the excitation light, the illumination intensity of the annular light field is absolutely consistent, the larger illumination area during single-molecule sensing detection is provided, and the detection efficiency and accuracy are improved.
The transmission waveguide 22, the excitation waveguide body 31 and the core layer of the micro-ring resonant cavity 32 are all formed by preparing a film by a plasma enhanced chemical vapor deposition method and then performing dry etching.
Of course, the transmission waveguide 22 also has an evanescent field due to the difference in refractive index of the core and cladding layers.
In another embodiment, as shown in fig. 1, a nano-integrated optical chip for single molecule sensing may include multiple groups of excitation waveguide units 3 and two groups of optical input units 2, where the multiple groups of excitation waveguide units 3 are arranged parallel to each other on the surface of the fluorescent transmission layer 1, and a single excitation waveguide unit 3 includes a linear excitation waveguide body 31 and multiple micro-ring resonators 32 arranged beside the excitation waveguide body 31 and spaced along a direction parallel to the excitation waveguide body 31, where, of course, the side includes a case of single side arrangement and two side arrangements, and the number of micro-ring resonators 32 is greater when two side arrangements are provided, and the illumination area is larger. The plurality of groups of excitation waveguide units 3 have a plurality of mutually parallel linear excitation waveguide bodies 31; the transmission waveguides 22 in both sets of optical input units 2 have a multi-stage Y-shaped bifurcated structure.
The input end of the transmission waveguide 22 is a single interface and is connected with the coupling grating 21, the output end of the transmission waveguide 22 presents a branched multi-interface and is further connected with a plurality of linear excitation waveguide bodies 31, as shown in fig. 1, the multi-interface output end of one transmission waveguide 22 is connected with the upper end of the excitation waveguide body 31, and the multi-interface output end of the other transmission waveguide 22 is connected with the lower end of the excitation waveguide body 31, so that an excitation light illumination area which is uniform and consistent on a plurality of detection sites is provided.
As shown in fig. 5, the nano-integrated optical chip for single molecule sensing in this embodiment further includes an image sensing unit 5, where the image sensing unit 5 is disposed on a surface of the fluorescent transmission layer 1 on a side facing away from the excitation waveguide unit 3, and the image sensing unit 5 is disposed corresponding to the excitation waveguide unit 3 above the fluorescent transmission layer 1, and is configured to receive kinetic information of a single molecule to be detected in the sample 100 to be detected in the detection area. The image sensing unit 5 may be a CMOS sensor. Of course, the image sensing unit 5 does not exclude other types of sensors.
In one embodiment, the fluorescent light-transmitting layer 1 comprises a wavelength-selective layer 11, and the excitation waveguide unit 3 is arranged on the wavelength-selective layer 11. At this time, the detection area is located on the wavelength selective layer 11, that is, the excitation waveguide unit 3 is disposed on the wavelength selective layer 11, and the sample 100 to be detected for fluorescence detection is also added on the wavelength selective layer 11, so that the applied fluorescent label is excited by the evanescent field on the excitation waveguide unit 3, and emits fluorescence emission light. The wavelength selective layer 11 has selective permeability to fluorescence emission light of a specific wavelength band, selectively transmits fluorescence emission light, blocks stray light such as excitation light, and suppresses noise, thereby avoiding signal interference caused by transmission of excitation light, further enhancing suppression of excitation light without causing loss to fluorescence emission light.
In one embodiment, the material of the wavelength selective layer 11 is isotropic amorphous silicon carbide. The refractive index of amorphous silicon carbide ranges from 2.3 to 2.8, and the transmission spectrum is shown in fig. 7. The amorphous silicon carbide can block a part of excitation light and allow light in a wave band of fluorescence emission light to pass through, so that the transmission loss of the fluorescence emission light is effectively prevented. For example, fluorescence emission light having a wavelength of more than 500nm can be selectively transmitted, and excitation light having a wavelength of 500nm or less can be suppressed.
As another alternative embodiment, the material of the wavelength selective layer 11 is a photonic crystal material, the photonic crystal material is formed by alternately stacking a silicon layer and a silicon dioxide layer, the thickness ranges of the silicon layer and the silicon dioxide layer are 100nm-400nm, and the wavelength selective layer 11 can also block the excitation light and transmit the fluorescence emission light by using the photonic crystal material.
The fluorescent light-transmitting layer 1 in the present embodiment further includes a fluorescent guiding layer 12 on the basis that the above-described fluorescent light-transmitting layer 1 includes the wavelength-selective layer 11. The fluorescence guide layer 12 is arranged between the wavelength selective layer 11 and the excitation waveguide unit 3, i.e. the excitation waveguide unit 3 is arranged on the fluorescence guide layer 12, and the detection area is also arranged on the surface of the fluorescence guide layer 12. The fluorescence guide layer 12 enables fluorescence emission light to be transmitted as much as possible and concentrated to the wavelength selective layer 11 to reach the image sensing unit 5.
Specifically, the thickness of the fluorescent guiding layer 12 is in the range of 1um to 5um. The thickness of the fluorescent guiding layer 12 is used for controlling the distance between the plane of the excitation waveguide unit 3 and the image sensing unit 5, and an excessive distance can cause signal interference when an evanescent field around the excitation waveguide body 31 and the micro-ring resonant cavity 32 enters the image sensing unit 5, and an excessive distance can cause a larger loss when fluorescent emitted light around the excitation waveguide body 31 and the micro-ring resonant cavity 32 reaches the image sensing unit 5.
The material of the fluorescent guiding layer 12 in this embodiment is silica, and the refractive index is in the range of 1.3-1.6.
Of course, in another embodiment, it is also possible to provide that the fluorescence transmission layer 1 comprises only the wavelength selective layer 11, the distance between the excitation waveguide unit 3 and the image sensing unit 5 being adjusted by the thickness of the wavelength selective layer 11.
In this embodiment, for parallel detection, 10 may be provided in a single excitation waveguide unit 3 2 To 10 4 A number of micro-ring resonators 32, for example, 100, 1000, 10000; the number of micro-ring resonators 32 that can be provided on a single chip is 10 3 To 10 7 For example, 1000, 10000, 100000, 1000000, 10000000 are provided.
The embodiment of the invention also provides a fluorescence detection method, as shown in fig. 8 and 9, which is applied to the nano integrated optical chip for single molecule sensing, and comprises the following steps:
s1, adding a sample 100 to be detected into a detection area of the fluorescent transmission layer 1, wherein the sample 100 to be detected comprises a single molecule 101 to be detected.
The fluorescence detection method of this example is used to detect kinetic information of single molecules on the surface of or in the cell membrane. Thus, taking the sample 100 to be tested of the present embodiment as a cell, the cell membrane surface of the cell contains the single molecule 101 to be tested. In fact, added to the detection zone is an aqueous solution 300 to be tested comprising cells and cell culture medium, followed by incubation at 37 ℃ so that the cells as sample 100 to be tested form an adherent surface on the surface of the nano-integrated optical chip for single molecule sensing, the cell membrane being in close contact with the surface of the nano-integrated optical chip for single molecule sensing, including in close contact with the surface of the excitation waveguide unit 3.
For example, when the fluorescent light-transmitting layer 1 includes the fluorescent light-guiding layer 12 and the wavelength-selective layer 11, the fluorescent light-guiding layer 12 is made of silica, and the cladding of the excitation waveguide body 31 and the micro-ring resonator 32 of the excitation waveguide unit 3 is also made of silica, the cell membrane of the cell is closely adhered to the surface of the silica layer, and the cell as the sample 100 to be measured is fixed on the surface of the detection region.
S2, adding a binding molecule 200 containing fluorescent labels into the detection area of the fluorescent transmission layer 1, wherein the binding molecule 200 is combined with the single molecule 101 to be detected.
As shown in FIG. 8, a reaction solution containing a fluorescent-labeled binding molecule 200 is added, wherein the fluorescent-labeled binding molecule 200 is capable of specifically binding to a single molecule 101 to be detected on the surface of a cell membrane, and the fluorescent-labeled binding molecule 200 is immobilized in a detection region.
And S3, controlling the nano integrated optical chip for single-molecule sensing to emit excitation light so that the fluorescent label is excited to emit fluorescence emission light, and the fluorescence emission light is emitted from the fluorescence transmission layer 1.
By capturing the fluorescent signal emitted when the binding molecule 200 with fluorescent label is excited, the binding or dissociation event between the single molecule 101 to be detected and the binding molecule 200 is discriminated, and the kinetic information of the interaction of the two single molecules is obtained.
Specifically, the diffusion speed of the free fluorescent molecules added to the detection area is constant, the residence time of the free fluorescent molecules is microsecond, and the light emitting time is very short; in this application, the binding molecule 200 with fluorescent label is specifically bound with the single molecule 101 to be detected in the chip detection area and is in a fixed state, the residence time of the fluorescent molecule in the fixed state is in millisecond level and long in light emitting time without being influenced by the volume of the excitation area 4, that is, the free fluorescent molecule is smaller by several orders of magnitude than the light emitting time of the fluorescent molecule in the fixed state, the smaller the volume of the excitation area 4 is, the shorter the residence time of the free fluorescent molecule is, the free fluorescent molecule is more easily shielded as background noise, so that the evanescent field can efficiently sense the interaction process of the binding and the dissociation of the detection single molecule.
In a preferred embodiment, a single cell in the sample 100 to be tested covers at least one micro-ring resonator 32, i.e., cells in the sample 100 to be tested span one or two or more micro-ring resonators 32. The annular light field on the space of the micro-ring resonant cavity 32 improves the area of single cells illuminated by the excitation light from the same excitation waveguide body 31, improves the intensity consistency of the excitation light in local areas of the cells and the total area of illumination, and further improves the detection efficiency and accuracy.
In one embodiment, the fluorescence detection method further comprises: an image sensing unit 5 is arranged on a surface of the fluorescent light-transmitting layer 1 facing away from the excitation waveguide unit 3, the image sensing unit 5 being adapted to receive fluorescent emitted light.
In the above description, technical details of patterning, etching, and the like of each layer are not described in detail. Those skilled in the art will appreciate that layers, regions, etc. of the desired shape may be formed by a variety of techniques. In addition, to form the same structure, those skilled in the art can also devise methods that are not exactly the same as those described above. In addition, although the embodiments are described above separately, this does not mean that the measures in the embodiments cannot be used advantageously in combination.
Although embodiments of the present invention have been described in connection with the accompanying drawings, various modifications and variations may be made by those skilled in the art without departing from the spirit and scope of the invention, and such modifications and variations fall within the scope of the invention as defined by the appended claims.
Claims (14)
1. A nano-integrated optical chip for single molecule sensing, comprising:
a fluorescence transmission layer (1), wherein the surface of the fluorescence transmission layer (1) is provided with a detection area;
at least two groups of light input units (2), wherein the input end of any one of the light input units (2) is connected with an external laser light source, and the output end of the light input unit extends into the detection area so as to introduce excitation light into the detection area;
at least one set of excitation waveguide units (3) arranged on the fluorescent transmission layer (1) and located in the detection area; the excitation waveguide unit (3) comprises an excitation waveguide body (31) and a plurality of micro-ring resonant cavities (32), and two ends of the excitation waveguide body (31) are respectively connected with the output ends of the two groups of light input units (2) so as to bidirectionally input the excitation light; the micro-ring resonant cavities (32) are arranged at intervals beside the excitation waveguide body (31) along the direction parallel to the excitation waveguide body (31); the micro-ring resonant cavity (32) is in a ring shape, the distance between the center of any micro-ring resonant cavity (32) and the center of the excitation waveguide body (31) is smaller than half of the wavelength of the excitation light along the direction perpendicular to the excitation waveguide body (31), and the circumference of the micro-ring resonant cavity (32) is set to be an integral multiple of the wavelength of the excitation light.
2. The nano-integrated optical chip for single molecule sensing according to claim 1, wherein the optical input unit (2) comprises a coupling grating (21) and a transmission waveguide (22);
one end of the coupling grating (21) is connected with an external laser light source, the other end of the coupling grating (21) is connected with one end of the transmission waveguide (22), and the other end of the transmission waveguide (22) is connected with the excitation waveguide body (31).
3. The nano-integrated optical chip for single molecule sensing according to claim 2, wherein the material of the transmission waveguide (22), the excitation waveguide body (31) and the micro-ring resonator (32) is the same, and the transmission waveguide (22), the excitation waveguide body (31) and the micro-ring resonator (32) each comprise a core layer and a cladding layer, and the refractive index of the core layer is larger than the refractive index of the cladding layer.
4. The nano-integrated optical chip for single-molecule sensing according to claim 3, wherein the material of the core layer is amorphous silicon carbide, and the refractive index of the core layer ranges from 2.3 to 3.0; the cladding is made of silicon dioxide or aqueous solution;
the outer surface of the micro-ring resonant cavity (32) is formed with an excitation region (4), and the thickness of the excitation region (4) is 50nm.
5. The nano-integrated optical chip for single molecule sensing as claimed in claim 2, wherein,
a plurality of groups of excitation waveguide units (3) are arranged in parallel;
the transmission waveguides (22) in the two groups of the optical input units (2) have a multi-stage Y-shaped bifurcated structure.
6. The nano-integrated optical chip for single molecule sensing of any one of claims 1-5, further comprising:
and the image sensing unit (5) is arranged on the surface of the fluorescence transmission layer (1) at the side away from the excitation waveguide unit (3).
7. The nano-integrated optical chip for single molecule sensing according to claim 6, wherein the fluorescent transmission layer (1) comprises a wavelength selective layer (11);
the excitation waveguide unit (3) is arranged on the wavelength selective layer (11).
8. The nano-integrated optical chip for single molecule sensing according to claim 7, wherein the material of the wavelength selective layer (11) is isotropic amorphous silicon carbide.
9. The nano-integrated optical chip for single molecule sensing according to claim 7, wherein the material of the wavelength selective layer (11) is a photonic crystal material consisting of silicon layers and silicon dioxide layers alternately, the thickness of the silicon layers and the silicon dioxide layers being in the range of 100nm-400nm.
10. The nano-integrated optical chip for single molecule sensing according to claim 7, wherein the fluorescent light-transmitting layer (1) further comprises a fluorescent guiding layer (12);
the fluorescence guiding layer (12) is arranged between the wavelength selection layer (11) and the excitation waveguide unit (3), the excitation waveguide unit (3) is arranged on the fluorescence guiding layer (12), and the detection area is arranged on the surface of the fluorescence guiding layer (12).
11. The nano-integrated optical chip for single molecule sensing according to claim 10, wherein the thickness of the fluorescent guiding layer (12) ranges from 1um to 5um.
12. A fluorescence detection method applied to the nano-integrated optical chip for single-molecule sensing according to any one of claims 1 to 11, comprising:
adding a sample (100) to be detected into the detection area of the fluorescent transmission layer (1), wherein the sample (100) to be detected comprises single molecules (101) to be detected;
adding a binding molecule (200) comprising a fluorescent label into the detection region of the fluorescent transmission layer (1), the binding molecule (200) being bound to the single molecule (101) to be detected;
the nano-integrated optical chip for single-molecule sensing is controlled to emit excitation light, so that the fluorescent label is excited to emit fluorescence emission light, and the fluorescence emission light is emitted from the fluorescence transmission layer (1).
13. The fluorescence detection method according to claim 12, wherein the sample (100) to be detected covers at least one of the micro-ring resonators (32).
14. The fluorescence detection method of claim 13, further comprising:
an image sensing unit (5) is arranged on a surface of the fluorescence transmission layer (1) facing away from the excitation waveguide unit (3), the image sensing unit (5) being adapted to receive the fluorescence emission light.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410039508.6A CN117554349B (en) | 2024-01-11 | 2024-01-11 | Nano integrated optical chip for single molecule sensing and fluorescence detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410039508.6A CN117554349B (en) | 2024-01-11 | 2024-01-11 | Nano integrated optical chip for single molecule sensing and fluorescence detection method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117554349A CN117554349A (en) | 2024-02-13 |
| CN117554349B true CN117554349B (en) | 2024-03-19 |
Family
ID=89821943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410039508.6A Active CN117554349B (en) | 2024-01-11 | 2024-01-11 | Nano integrated optical chip for single molecule sensing and fluorescence detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117554349B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20070018083A (en) * | 2004-05-27 | 2007-02-13 | 쓰리엠 이노베이티브 프로퍼티즈 컴파니 | Dielectric microcavity fluorosensors excited with a broadband light source |
| CN103682966A (en) * | 2013-12-02 | 2014-03-26 | 北京工业大学 | Living cell bio-laser based on hollow-core photonic crystal fiber carrier |
| CN108375556A (en) * | 2018-01-16 | 2018-08-07 | 南京大学 | A kind of new type of high sensitivity and the unmarked Terahertz sensor for measuring monolayer |
| CN111426450A (en) * | 2020-03-17 | 2020-07-17 | 天津大学 | Resonant cavity enhanced monolithic integrated sensor and measurement method |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007137060A2 (en) * | 2006-05-16 | 2007-11-29 | Applied Biosystems | Systems, methods, and apparatus for single molecule sequencing |
-
2024
- 2024-01-11 CN CN202410039508.6A patent/CN117554349B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20070018083A (en) * | 2004-05-27 | 2007-02-13 | 쓰리엠 이노베이티브 프로퍼티즈 컴파니 | Dielectric microcavity fluorosensors excited with a broadband light source |
| CN103682966A (en) * | 2013-12-02 | 2014-03-26 | 北京工业大学 | Living cell bio-laser based on hollow-core photonic crystal fiber carrier |
| CN108375556A (en) * | 2018-01-16 | 2018-08-07 | 南京大学 | A kind of new type of high sensitivity and the unmarked Terahertz sensor for measuring monolayer |
| CN111426450A (en) * | 2020-03-17 | 2020-07-17 | 天津大学 | Resonant cavity enhanced monolithic integrated sensor and measurement method |
Non-Patent Citations (2)
| Title |
|---|
| Label-Free Optical Single-Molecule Micro- and Nanosensors;Sivaraman Subramanian等;《Adv. Mater. 》;20181231;第1-21页 * |
| 倏逝场照明的集成零模波导纳米孔芯片;俞鹏飞等;《光学精密工程》;20220131;第62-70页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117554349A (en) | 2024-02-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Cai et al. | Overview of the coupling methods used in whispering gallery mode resonator systems for sensing | |
| US7903240B2 (en) | Optical sensing device | |
| US7512298B2 (en) | Optical sensing methods | |
| US7920267B2 (en) | Micro integrated planar optical waveguide type SPR sensor | |
| US9063135B2 (en) | Method for chip-integrated label-free detection and absorption spectroscopy with high throughput, sensitivity, and specificity | |
| US8195014B2 (en) | Optical chemical detector and method | |
| US8506887B2 (en) | Porous membrane waveguide sensors and sensing systems therefrom for detecting biological or chemical targets | |
| US7314751B2 (en) | Fluorescence detection system including a photonic band gap structure | |
| US7304734B2 (en) | Fluorescence analysis optical multiplexer/demultiplexer, fluorescence analysis optical module, fluorescence analyzer, fluorescence/photothermal conversion spectroscopic analyzer, and fluorescence analysis chip | |
| CN109581584B (en) | Silicon-lithium niobate heterogeneous integration scanning chip and preparation method and application thereof | |
| US9693715B2 (en) | Optical sensor for detecting chemical, biochemical or biological substances | |
| US5745231A (en) | Method of fluorescence analysis comprising evanescent wave excitation and out-of-plane photodetection | |
| US20120012739A1 (en) | Optical microresonator system | |
| Hua et al. | Integrated optical fluorescence multisensor for water pollution | |
| EP2053385A2 (en) | Apparatus and method for detecting the presence of an agent | |
| KR20050071567A (en) | Sensor | |
| US7574089B1 (en) | Optofluidic devices and methods of using the same | |
| CN116026790A (en) | Sensor based on continuous domain constraint state sub-wavelength grating runway type resonant cavity | |
| CN117554349B (en) | Nano integrated optical chip for single molecule sensing and fluorescence detection method | |
| JP2015078866A (en) | Optical resonator structure | |
| Eustache et al. | Miniaturized Bloch surface wave platform on a multicore fiber | |
| US20240094204A1 (en) | Photonic biosensors for multiplexed diagnostics and a method of use | |
| US20060023216A1 (en) | Integrated luminescence read device | |
| KR20060089103A (en) | Biochip flatform using a spherical cavity resonator and biochemical sensor having the same | |
| JP2024084468A (en) | Optical sensing circuit and optical sensing system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |