CN111781184B - Photon nanometer jet, laser array unit and single-molecule fluorescent gene sequencer - Google Patents
Photon nanometer jet, laser array unit and single-molecule fluorescent gene sequencer Download PDFInfo
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Abstract
The photon nanometer jet provided by the invention is composed of medium materials with different refractive indexes, parallel light is incident to the bottom end of the photon nanometer jet, the effect similar to zero-mode waveguide can be realized, the ultra-narrow optical field local area is generated by the highly local optical field distribution obtained at the other end, the observation volume of the normal biological reaction is achieved, the photon nanometer jet can be used for single molecule fluorescence excitation, the method for realizing single molecule real-time nucleic acid sequencing is used for improving the flux in the single molecule real-time sequencing process and reducing the complexity of an optical system. In addition, the invention also provides a laser array unit comprising the photon nanometer jet and a monomolecular fluorescent gene sequencer.
Description
Technical Field
The invention relates to a single-molecule fluorescence real-time imaging technology, in particular to a photon nanometer jet and laser array unit and a single-molecule fluorescence gene sequencer.
Background
The gene sequencer provides an important means for researching gene structure, replication, transcription, translation, expression regulation and control, physiological functions of expression products, cell signal transduction and other life phenomena. The first generation Sanger sequencing method needs multiple groups of primers, can only detect common mutant consensus sequences, and cannot realize the detection of the mutation level of a single virus molecule. The throughput of the second-generation sequencing is improved by thousands of times, the sequencing cost is greatly reduced, and the sequencing result has high accuracy, and mainly comprises a pyrosequencing technology of Roche company, a Solexa sequencing technology of Illumina company and a SOLID sequencing technology of ABI company. Second generation technologies are mainly characterized by high throughput and low cost, but second generation sequencing is typically less than 300bp in length and sequencing coverage is affected by GC components.
The third generation gene sequencing is characterized by Single molecule, long read length and amplification-free, and represents the Single-molecule Real-time (SMRT) sequencing technology of RS (Real-time sequencing) of product PacBio. The SMRT sequencing technique relies on the fact that nucleic acids bind to long strands of DNA at the polymerase, which takes much longer to form chemical bonds than other free bases. By counting the time length of the fluorescence signal, the bound base and the free base can be distinguished, and the sequence measurement of the DNA long chain is realized.
An array structure based on zero mode waveguides is proposed in patent CN101914620B, which realizes an optical confinement array. The zero mode waveguide can achieve the prevention of electromagnetic radiation having a frequency below a cutoff frequency from propagating longitudinally through the core of the zero mode waveguide, wherein the zero mode waveguide creates an effective observation volume of resolvable single molecules upon illumination of the zero mode waveguide with electromagnetic radiation having a frequency below the cutoff frequency. Chip surface density reaches 4 x 104 optical confinement arrays of confinement per square millimeter. However, due to the size limitation of the zero mode waveguide structure and the depth of the nanopore, the filling rate is low during the sequencing process, and only about 30% of the zero mode waveguide is successfully biologically modified, so that single molecule sequencing can be performed. The pore structure reduces the throughput in the actual sequencing process, increasing the cost of the test. The average read length during sequencing of the RSII series by Pacbio was about 4600bp, throughput was about 3.2Gb, run time was 40 hours, accuracy was 82.1% -84.9%, and run cost was $ 4800 and data cost was $ 1500/Gb. However, the zero mode waveguide technology of PacBio cannot greatly increase the loading rate of the DNA fragments in the nanopore array, which is only about 30% at present. The instrument cost and the sequencing use cost are much higher than those of the second-generation sequencing, and the market share is limited.
Disclosure of Invention
In view of the above, there is a need to provide a photonic nanojet that can improve throughput and reduce complexity of optical systems in real-time sequencing of single molecules, which is a drawback of the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a photonic nanojet comprising a core, a cladding layer surrounding the core and a top layer disposed on top of the core and the cladding layer, wherein:
the core layer has a diameter of 0.9-1.1 μm, a refractive index of 1.4-1.7, the cladding layer has a thickness of 0.45-0.55 μm, a refractive index of 1.3-1.6, the top layer has a length of 90-110 nm, a thickness of 0.45-0.55 μm, and a refractive index of 2.25-2.75.
In some of these embodiments, the core layer is barium crown glass, the cladding layer is silicon dioxide, and the top layer is titanium dioxide.
In some of these embodiments, the top layer is conical.
The invention also provides photon nanometer spraying, which comprises a core layer, a cladding layer for coating the periphery of the core layer, a top layer arranged on the core layer, and metal film layers arranged on the two sides of the top layer and in contact with the core layer and the cladding layer, wherein:
the core layer has a diameter of 0.9-1.1 μm, a refractive index of 1.4-1.7, the cladding layer has a thickness of 0.45-0.55 μm, a refractive index of 1.3-1.6, the top layer has a length and width of 90-110 nm, a refractive index of 2.25-2.75, and the metal film layer has a thickness of 34-66 nm.
In some embodiments, the core layer is barium crown glass, the cladding layer is silicon dioxide, the top layer is titanium dioxide, and the metal thin film layer is aluminum.
The invention also provides photon nanometer spraying, which comprises a cladding, a top layer arranged on the cladding, wherein a metal thin film layer is arranged on the two side edges of the top layer and the part contacted with the cladding, the thickness of the cladding is between 0.45 and 0.55 mu m, the refractive index is between 1.3 and 1.6, the length and the width of the top layer are between 90 and 110nm, the refractive index is between 2.25 and 2.75, and the thickness of the metal thin film layer is between 34 and 66 nm.
In some of these embodiments, the cladding layer is silica, the top layer is titania, and the metal film layer is aluminum.
The invention also provides a laser array unit which comprises a plurality of photon nano-jets, and the photon nano-jets are arranged in an array.
The invention also provides a single-molecule fluorescent gene sequencer which comprises the laser array unit.
The invention adopts the technical scheme that the method has the advantages that:
the photon nanometer jet provided by the invention is composed of medium materials with different refractive indexes, parallel light is incident to the bottom end of the photon nanometer jet, the effect similar to zero-mode waveguide can be realized, the ultra-narrow optical field local area is generated by the highly local optical field distribution obtained at the other end, the observation volume of the normal biological reaction is achieved, the photon nanometer jet can be used for single molecule fluorescence excitation, the method for realizing single molecule real-time nucleic acid sequencing is used for improving the flux in the single molecule real-time sequencing process and reducing the complexity of an optical system.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic structural diagram of a photonic nanojet provided in example 1 of the present invention;
fig. 2 is a schematic structural diagram of a laser array unit provided in embodiment 1 of the present invention;
FIG. 3 is an optical schematic diagram of photonic nanojet provided in example 1 of the present invention;
FIG. 4 is a schematic structural diagram of a photonic nanojet provided in example 2 of the present invention;
fig. 5 is a schematic structural diagram of a laser array unit provided in embodiment 2 of the present invention;
FIG. 6 is a schematic structural diagram of a photonic nanojet provided in example 3 of the present invention;
fig. 7 is a schematic structural diagram of a laser array unit according to embodiment 3 of the present invention;
FIG. 8 is a schematic structural diagram of a photonic nanojet provided in example 4 of the present invention;
fig. 9 is a schematic structural diagram of a photonic nanojet provided in embodiment 4 of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious 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 1
Referring to fig. 1, a schematic structural diagram of a photonic nanojet 100 provided in embodiment 1 of the present invention includes a core layer 110, a cladding layer 120 wrapping a periphery of the core layer 110, and a top layer 130 disposed on top of the core layer 110 and the cladding layer 120.
Furthermore, the diameter of the core layer 110 is between 0.9 and 1.1 μm, the refractive index is between 1.4 and 1.7, the thickness of the cladding layer 120 is between 0.45 and 0.55 μm, the refractive index is between 1.3 and 1.6, the length of the top layer 130 is between 90 and 110nm, the thickness is between 0.45 and 0.55 μm, and the refractive index is between 2.25 and 2.75.
In some embodiments, the core layer is barium crown glass, the cladding layer is silicon dioxide, the top layer is titanium dioxide, and the top layer is conical.
Referring to fig. 2, the present embodiment further provides a laser array unit 10, which includes a plurality of the photon nano-jets 100, and the photon nano-jets 100 are arranged in an array.
In some of these embodiments, the array is arranged from the transverse direction to the longitudinal direction by 2 x 103Total number of 4 x 106The length and width of the array were 4 mm.
It can be understood that the photon nano-jet provided by the embodiment adopts the planar convex structure, so that the problem of low filling rate of the pore structure is effectively solved, the chip with the unit array is provided, and a foundation is provided for sequencing with higher flux.
Referring to fig. 3, a schematic diagram of an optical path of a photonic nano-jet according to an embodiment of the present invention is shown, when parallel light is incident from the bottom of the photonic nano-jet, because the photonic nano-jet is made of dielectric materials with different refractive indexes, the parallel light is incident to the bottom of the structure, and a highly localized light field distribution is obtained at the other end, and the highly localized light field distribution can be achieved due to near-field diffraction and phase superposition, so that an observation volume of a biological normal reaction is achieved, a condition that a single molecule is excited to emit fluorescence each time can be satisfied, a photonic nano-jet with a transverse full width at half maximum of about 100nm and a longitudinal dimension of about 200nm can be obtained, and the photonic nano-jet can be used for single-molecule fluorescence excitation, a single-molecule real-time nucleic acid sequencing is achieved, and a flux in a single-molecule real-time sequencing process is increased and a complexity of an optical system is reduced.
In addition, the incident light field of the photon nanometer jet provided by the invention only needs to be parallel light, so that a complex system of laser beam splitting is avoided, and the difficulty of an optical system is reduced.
Example 2
Referring to fig. 4, a schematic structural diagram of a photonic nanojet 200 provided in this embodiment 2 includes a core layer 210, a cladding layer 220 covering the periphery of the core layer 210, a top layer 230 disposed on the core layer 210, and a metal thin film layer 240 disposed on two sides of the top layer 230 and in contact with the core layer 210 and the cladding layer 220.
Further, the diameter of the core layer 210 is between 0.9 μm and 1.1 μm, the refractive index is between 1.4 and 1.7, the thickness of the cladding layer 220 is between 0.45 μm and 0.55 μm, the refractive index is between 1.3 and 1.6, the length and width of the top layer 230 are between 90nm and 110nm, the refractive index is between 2.25 and 2.75, and the thickness of the metal thin film layer 240 is between 34nm and 66 nm.
In some of these embodiments, the core layer is barium crown glass, the cladding layer is silicon dioxide, and the top layer is titanium dioxide.
Referring to fig. 5, the present embodiment 2 further provides a laser array unit 20, which includes a plurality of the photon nano-jets 200, and the photon nano-jets 200 are arranged in an array.
In some of these embodiments, the array is arranged from the transverse direction to the longitudinal direction by 2 x 103Total number of 4 x 106The length and width of the array were 4 mm.
When parallel light is incident from the bottom of the photon nanometer jet, the photon nanometer jet is made of medium materials with different refractive indexes, the parallel light is incident to the bottom end of the structure, high-local-area light field distribution can be obtained at the other end, the high-local-area light field can be realized due to near-field diffraction and phase superposition, the observation volume of the normal biological reaction is achieved, the condition that a single molecule is excited to emit fluorescence every time can be met, the photon nanometer jet with the transverse full width at half maximum of about 100nm and the longitudinal dimension of about 200nm can be obtained, the photon nanometer jet can be used for single-molecule fluorescence excitation, single-molecule real-time nucleic acid sequencing is realized, and the flux in the single-molecule real-time sequencing process is improved and the complexity of an optical system is reduced.
Example 3
Referring to fig. 6, a schematic structural diagram of a photon nano-jet 300 provided in this embodiment 3 includes a cladding 310, a top layer 320 disposed on the cladding 310, and a metal thin film layer 330 disposed on two side edges of the top layer 320 and contacting with the cladding, wherein the thickness of the cladding 310 is 0.45 μm to 0.55 μm, the refractive index is 1.3 to 1.6, the length and width of the top layer 320 are 90nm to 110nm, the refractive index is 2.25 to 2.75, and the thickness of the metal thin film layer 330 is 34nm to 66 nm.
In some of these embodiments, the cladding layer is silica, the top layer is titania, and the metal film layer is aluminum.
Referring to fig. 7, embodiment 3 further provides a laser array unit 30, which includes a plurality of the photon nano-jets 300, and the photon nano-jets 300 are arranged in an array.
In some of these embodiments, the array is arranged from the transverse direction to the longitudinal direction by 2 x 103Total number of 4 x 106The length and width of the array were 4 mm.
When parallel light is incident from the bottom of the photon nanometer jet, the photon nanometer jet is made of medium materials with different refractive indexes, the parallel light is incident to the bottom end of the structure, high-local-area light field distribution can be obtained at the other end, the high-local-area light field can be realized due to near-field diffraction and phase superposition, the observation volume of the normal biological reaction is achieved, the condition that a single molecule is excited to emit fluorescence every time can be met, the photon nanometer jet with the transverse full width at half maximum of about 100nm and the longitudinal dimension of about 200nm can be obtained, the photon nanometer jet can be used for single-molecule fluorescence excitation, single-molecule real-time nucleic acid sequencing is realized, and the flux in the single-molecule real-time sequencing process is improved and the complexity of an optical system.
Example 4
Fig. 8 and 9 are schematic views of a photonic nano-jet structure according to embodiment 4 of the present invention.
The photonic nanojet provided in fig. 8 consists of two refractive index dielectric materials, the middle 410 being a cylindrical structure with a refractive index of 1.46, a diameter of 2.5 μm, the material being silicon dioxide; the upper and lower parts (420) are made of high-refractive-index materials, the refractive index is 2.3, the width is 320nm, the thickness is 50nm, and the materials are niobium oxide.
The structure in fig. 9 is also composed of two refractive index dielectric materials, the middle 510 is a square structure with a refractive index of 1.46, the side length is 1 μm, and the material is silicon dioxide; the upper and lower parts (520) are made of high-refractive-index materials, the refractive index is 2.3, the width is 250nm, the thickness is 50nm, and the materials are niobium oxide.
When parallel light is incident from the bottom of the photon nanometer jet, the photon nanometer jet is made of medium materials with different refractive indexes, the parallel light is incident to the bottom end of the structure, high-local-area light field distribution can be obtained at the other end, the high-local-area light field can be realized due to near-field diffraction and phase superposition, the observation volume of the normal biological reaction is achieved, the condition that a single molecule is excited to emit fluorescence every time can be met, the photon nanometer jet with the transverse full width at half maximum of about 100nm and the longitudinal dimension of about 200nm can be obtained, the photon nanometer jet can be used for single-molecule fluorescence excitation, single-molecule real-time nucleic acid sequencing is realized, and the flux in the single-molecule real-time sequencing process is improved and the complexity of an optical system is reduced.
The photon nanometer jet provided by the embodiment of the invention utilizes the near-field diffraction and the phase superposition effect of the dielectric material under the micro-nano size, realizes the height local area of the optical field, the transverse dimension is compressed to 100nm, and the longitudinal dimension is about 200 nm; the method not only achieves the observation volume of the normal biological reaction, but also can meet the condition of exciting single molecules to emit fluorescence each time, can be used in a single-molecule fluorescent gene sequencer and can carry out single-molecule sequencing in real time.
Of course, the photonic nanojet of the present invention can have various changes and modifications, and is not limited to the specific structure of the above embodiment. In conclusion, the scope of the present invention should include those changes or substitutions and modifications which are obvious to those of ordinary skill in the art.
Claims (8)
1. A photon nanometer jet device for single molecule imaging, which is characterized by comprising a core layer, a cladding layer for cladding the periphery of the core layer and a top layer arranged on the top of the core layer and the cladding layer, wherein:
the core layer has a diameter of 0.9-1.1 μm, a refractive index of 1.4-1.7, the cladding layer has a thickness of 0.45-0.55 μm, a refractive index of 1.3-1.6, the top layer has a length of 90-110 nm, a thickness of 0.45-0.55 μm, and a refractive index of 2.25-2.75; the top layer is conical.
2. The photonic nanojet device for single molecule imaging of claim 1, wherein the core layer is barium crown glass, the cladding layer is silicon dioxide, and the top layer is titanium dioxide.
3. The photon nanometer injection device for single molecule imaging is characterized by comprising a core layer, a cladding layer for cladding the periphery of the core layer, a top layer arranged on the core layer, two side edges of the top layer and metal thin film layers arranged at the positions where the core layer is contacted with the cladding layer, wherein:
the core layer has a diameter of 0.9-1.1 μm, a refractive index of 1.4-1.7, the cladding layer has a thickness of 0.45-0.55 μm, a refractive index of 1.3-1.6, the top layer has a length and width of 90-110 nm, a refractive index of 2.25-2.75, and the metal film layer has a thickness of 34-66 nm.
4. The photonic nanojet device for single molecule imaging of claim 3, wherein the core layer is barium crown glass, the cladding layer is silica, the top layer is titanium dioxide, and the metal thin film layer is aluminum.
5. The photon nanometer injection device for single molecule imaging is characterized by comprising a cladding, a top layer arranged on the cladding, wherein a metal thin film layer is arranged on the two side edges of the top layer and in contact with the cladding, the thickness of the cladding is 0.45-0.55 mu m, the refractive index is 1.3-1.6, the length and the width of the top layer are 90-110 nm, the refractive index is 2.25-2.75, and the thickness of the metal thin film layer is 34-66 nm.
6. The photonic nanojet device for single molecule imaging of claim 5, wherein the cladding layer is silica, the top layer is titania, and the metal thin film layer is aluminum.
7. A laser array unit, comprising a plurality of photonic nanojet devices for single molecule imaging according to any one of claims 1, 3 or 5, wherein the plurality of photonic nanojet devices are arranged in an array.
8. A single molecule fluorescent gene sequencer comprising the laser array unit of claim 7.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010766330.7A CN111781184B (en) | 2020-08-03 | 2020-08-03 | Photon nanometer jet, laser array unit and single-molecule fluorescent gene sequencer |
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| CN116297377B (en) * | 2023-05-22 | 2023-07-28 | 深圳高性能医疗器械国家研究院有限公司 | Single-molecule fluorescence detection chip, preparation method thereof and single-molecule fluorescence detector |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011005397A1 (en) * | 2009-06-17 | 2011-01-13 | University Of North Carolina At Charlotte | Focusing multimodal optical microprobe devices |
| KR20110047546A (en) * | 2009-10-30 | 2011-05-09 | 유창희 | Nano-jet generating apparatus and nano-jet generating method |
| CN102305782A (en) * | 2011-08-10 | 2012-01-04 | 浙江大学 | Method and device for analyzing fluorescent correlation spectroscopy based on medium microsphere |
| CN106680985A (en) * | 2017-03-23 | 2017-05-17 | 暨南大学 | Optical capturing and detecting device and method based on quantum photon nano-jet array |
| RU178616U1 (en) * | 2017-11-01 | 2018-04-13 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Сибирский государственный университет геосистем и технологий" (СГУГиТ) | Device for forming a photon stream |
| CN109073910A (en) * | 2016-03-24 | 2018-12-21 | 汤姆逊许可公司 | Equipment at least one focus beam formed from incident electromagnetic wave near region |
| WO2019063221A1 (en) * | 2017-09-26 | 2019-04-04 | Thomson Licensing | Device for deviating and focusing light |
| CN109844579A (en) * | 2016-10-21 | 2019-06-04 | 汤姆逊许可公司 | At least one sub-wavelength dimensions object is shielded from the device and method of incident electromagnetic wave |
| CN110927879A (en) * | 2019-11-04 | 2020-03-27 | 桂林电子科技大学 | Nano optical jet scanning probe based on optical fiber tweezers |
| CN111051237A (en) * | 2017-09-01 | 2020-04-21 | 汤姆逊许可公司 | Adhesive-free adhesion to dielectric material layers using nanojet microstructures |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10416383B2 (en) * | 2017-07-20 | 2019-09-17 | The Board Of Trustees Of The University Of Illinois | Spatial control of the optical focusing properties of photonic nanojets |
| CN110749580B (en) * | 2019-09-18 | 2022-05-10 | 东南大学 | Nanometer microarray near-field structure for monomolecular fluorescence limited-domain excitation |
-
2020
- 2020-08-03 CN CN202010766330.7A patent/CN111781184B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011005397A1 (en) * | 2009-06-17 | 2011-01-13 | University Of North Carolina At Charlotte | Focusing multimodal optical microprobe devices |
| KR20110047546A (en) * | 2009-10-30 | 2011-05-09 | 유창희 | Nano-jet generating apparatus and nano-jet generating method |
| CN102305782A (en) * | 2011-08-10 | 2012-01-04 | 浙江大学 | Method and device for analyzing fluorescent correlation spectroscopy based on medium microsphere |
| CN109073910A (en) * | 2016-03-24 | 2018-12-21 | 汤姆逊许可公司 | Equipment at least one focus beam formed from incident electromagnetic wave near region |
| CN109844579A (en) * | 2016-10-21 | 2019-06-04 | 汤姆逊许可公司 | At least one sub-wavelength dimensions object is shielded from the device and method of incident electromagnetic wave |
| CN106680985A (en) * | 2017-03-23 | 2017-05-17 | 暨南大学 | Optical capturing and detecting device and method based on quantum photon nano-jet array |
| CN111051237A (en) * | 2017-09-01 | 2020-04-21 | 汤姆逊许可公司 | Adhesive-free adhesion to dielectric material layers using nanojet microstructures |
| WO2019063221A1 (en) * | 2017-09-26 | 2019-04-04 | Thomson Licensing | Device for deviating and focusing light |
| RU178616U1 (en) * | 2017-11-01 | 2018-04-13 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Сибирский государственный университет геосистем и технологий" (СГУГиТ) | Device for forming a photon stream |
| CN110927879A (en) * | 2019-11-04 | 2020-03-27 | 桂林电子科技大学 | Nano optical jet scanning probe based on optical fiber tweezers |
Non-Patent Citations (3)
| Title |
|---|
| Refractive index less than two: photonic nanojets yesterday, today and tomorrow;BORIS S. LUK’YANCHUK 等;《OPTICAL MATERIALS EXPRESS》;20170511;第7卷(第6期);1820-1847 * |
| Side-lobes-controlled photonic nanojet with a horizontal graded-index microcylinder;HUAMING XING 等;《Optics Letters》;20180901;第43卷(第17期);4292-4295 * |
| Superenhanced photonic nanojet by core-shell microcylinders;Cheng-Yang Liu;《Physics Letters A》;20120420;第376卷;1856–1860 * |
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