CN107290314B - Fluorescence detection method and device for unmarked micro-nano particles - Google Patents
Fluorescence detection method and device for unmarked micro-nano particles Download PDFInfo
- Publication number
- CN107290314B CN107290314B CN201710467212.4A CN201710467212A CN107290314B CN 107290314 B CN107290314 B CN 107290314B CN 201710467212 A CN201710467212 A CN 201710467212A CN 107290314 B CN107290314 B CN 107290314B
- Authority
- CN
- China
- Prior art keywords
- metal film
- noble metal
- light
- micro
- nano particles
- 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
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 15
- 238000001917 fluorescence detection Methods 0.000 title claims abstract description 10
- 229910000510 noble metal Inorganic materials 0.000 claims abstract description 32
- 229910052751 metal Inorganic materials 0.000 claims abstract description 22
- 239000002184 metal Substances 0.000 claims abstract description 22
- 239000011521 glass Substances 0.000 claims abstract description 18
- 239000002245 particle Substances 0.000 claims abstract description 16
- 230000007935 neutral effect Effects 0.000 claims abstract description 12
- 230000003287 optical effect Effects 0.000 claims abstract description 12
- 230000010287 polarization Effects 0.000 claims abstract description 11
- 239000013598 vector Substances 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 5
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 claims description 3
- 239000007769 metal material Substances 0.000 claims description 2
- 230000005284 excitation Effects 0.000 claims 2
- 238000001514 detection method Methods 0.000 abstract description 9
- 239000000523 sample Substances 0.000 description 16
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 7
- 239000010931 gold Substances 0.000 description 7
- 229910052737 gold Inorganic materials 0.000 description 7
- 239000004793 Polystyrene Substances 0.000 description 3
- 229920002223 polystyrene Polymers 0.000 description 3
- 241000700605 Viruses Species 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000010970 precious metal Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
Images
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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- 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
- G01N21/648—Specially adapted constructive features of fluorimeters using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a fluorescence detection method and a fluorescence detection device for unmarked micro-nano particles, and belongs to the technical field of near-field optics. The method is characterized in that a total internal reflection structure is utilized to excite the surface plasma resonance of noble metal, micro-nano particles positioned on the surface of a noble metal film and the surface plasma resonance of the noble metal film generate restrained total internal reflection, and the transmitted light excites the fluorescent solution where the particles are positioned to emit light. The device comprises a laser, a polarization regulator, a light wave vector-metal film surface plasma coupler, a glass slide glass with one side surface plated with a noble metal film, a bottom-lacking sample groove, a fluorescence collecting lens, an optical neutral filter, a photoelectric detector, a processing circuit, a computer and a rotary table. The invention realizes the direct detection of the micro-nano particles without labels, and the concentration of the particles is directly characterized by the intensity of fluorescence.
Description
Technical Field
The invention belongs to the technical field of near-field optics, relates to the action of a micro-nano structure and an evanescent field, and particularly relates to a method and a device for detecting label-free micro-nano particles by exciting fluorescence through frustrated total internal reflection.
Background
The detection of the micro-nano particles is important in the field of biomedicine, for example, the typical size range of viruses is 20-400 nm, and the accurate detection of the substances can provide an important detection means for pestilence forecast, pestilence outbreak control, public safety protection and the like. However, because the size of the substance is small, the substance is difficult to directly detect, and a fluorescence labeling method is usually adopted, so that the method has outstanding advantages in detection sensitivity, but a specific molecular probe needs to be screened out, and when the particle to be detected is unknown, the screening of the probe is difficult. Label-free direct detection of such particles is an urgent need for biomedical clinics.
Disclosure of Invention
The invention aims to provide a label-free direct detection method and a label-free direct detection device for micro-nano particles.
The technical scheme of the invention is as follows:
a fluorescence detection method of unmarked micro-nano particles utilizes a total internal reflection structure to excite the surface plasma resonance of noble metals, the micro-nano particles positioned on the surface of a noble metal film and the surface plasma resonance of the noble metal film generate restrained total internal reflection, and the fluorescence solution where the particles are positioned is excited to emit light through the transmitted light.
The method is applied to a device which comprises a laser, a polarization regulator, a light wave vector-metal film surface plasma coupler, a glass slide glass with one side surface plated with a noble metal film, a bottom-lacking sample groove, a fluorescence collecting lens, an optical neutral filter, a photoelectric detector, a processing circuit, a computer and a rotary table; wherein the laser provides incident light for exciting surface plasmon resonance of the metal film; the polarization regulator regulates the polarization state of incident light emitted by the laser to be p light parallel to an incident plane, and then the p light is incident on the noble metal film of the glass slide through the light wave vector-metal film surface plasma coupler, and the coupler and the glass surface below the metal film in the glass slide are connected together through the refractive index matching material; directly placing the bottom-lacking sample tank on a metal film, sealing the bottom-lacking sample tank with the metal film to form a sample tank, and injecting untreated micro-nano particles and a fluorescent solution into the tank; fluorescence emitted from the sample cell is collected by a fluorescence collecting system, then is converged on a photoelectric detector after passing through an optical neutral filter, and an electric signal obtained after passing through a processing circuit is input into a computer; the optical neutral filter is used for setting the threshold value of the fluorescence entering the photoelectric detector; the light wave vector-metal film surface plasma coupler is used for matching light wave vectors with wave vectors of the noble metal film; the light wave vector-metal film surface plasma coupler is connected with the glass bottom of the sample groove plated with the metal film through a refractive index matching material and fixed on the rotating table; the computer controls the rotating platform to rotate continuously.
The thickness h of the noble metal is within the range of 35nm to 75 nm.
Exciting light excites the surface plasma resonance of the noble metal film through the light wave vector-metal film surface plasma coupler, and the exciting light and the sample groove filled with micro-nano particles to be detected are respectively positioned at two sides of the noble metal film. The wavelength of the exciting light is within the full width at half maximum of the absorption peak of the fluorescent solution, and the bandwidth of the exciting light depends on the diameter and the concentration of the particles to be measured.
The diameter range of the micro-nano particles to be detected is 30nm-1000nm, and the surfaces of the particles are made of non-metallic materials. The included angle between the incident light and the normal line of the metal film is an incident angle, and the incident angle is continuously changed within the range of 0-90 degrees by rotating the rotating table. The focus of the fluorescence collecting lens is located on the surface of the gold film or within a defocus range from one particle diameter of the metal film. The light source may be a single light source, or a plurality of light sources having the same incident angle, or a plurality of light sources separated from the same light source and having the same incident angle.
The invention realizes the direct detection of the micro-nano particles without marking, and the concentration of the particles is directly characterized by the intensity of fluorescence.
Drawings
FIG. 1 is a schematic view of a fluorescence detection method of unmarked micro-nano particles
In the figure: 1, a laser; 2, a polarization state adjuster; 3 a prism used for matching the light wave vector and the plasma wave vector on the surface of the metal film; 4 a metal film plated on the glass slide; 5, a bottom-lacking sample groove; 6 a fluorescence collection lens; 7 an optical neutral filter; 8, a photoelectric detector; 9 a processing circuit; 10, a computer; 11 rotating table.
Detailed Description
The following further describes a specific embodiment of the present invention with reference to the drawings and technical solutions.
Examples
A fluorescence detection method and device for unmarked polystyrene particles with the diameter of 60nm are characterized in that a total internal reflection structure is utilized to excite the surface plasma resonance of noble metal, micro-nano particles positioned on the surface of a noble metal film generate restrained total internal reflection after the resonance action of the micro-nano particles and the surface plasma thereof, and transmitted light excites Fr633(sigma company) fluorescent substances with the absorption peak of 633nm in solution around the particles to emit light, the method is applied to the following devices, and the device comprises the following components: the device comprises a laser 1, a polarization regulator 2, a prism 3 for matching light wave vectors and metal film surface plasma wave vectors, a precious metal film 4 plated on a glass slide glass, a bottom-lacking sample groove 5, a fluorescence collecting lens 6, an optical neutral filter 7, a photoelectric detector 8, a processing circuit 9, a computer 10 and a rotary table 11; wherein, the light with central wavelength of 633nm emitted by the He-Ne laser 1 passes through the polarization regulator 2 to regulate the polarization state of the incident light into p light parallel to the incident surface, and then the p light is incident on the gold film 4 through the prism 3 to excite the surface plasma resonance; the sample groove 5 with the bottom is directly placed on the gold film and sealed with the gold film to form a sample groove, and untreated polystyrene particles and fluorescent solution are injected into the sample groove; fluorescence emitted from the sample cell is collected by a fluorescence collecting lens 6, then is converged on a photoelectric detector PMT 8 by an optical neutral filter 7, and a signal obtained after processing by a processing circuit 9 is input into a computer 10; the optical neutral filter 7 is used for setting the threshold value of the fluorescence entering the photoelectric detector; the prism 3 is connected with the bottom of the glass slide of the sample groove through the refractive index matching resin oil and fixed on the rotating platform 11; the computer 10 controls the turntable to rotate continuously.
The laser and the sample groove filled with polystyrene nano particles to be detected are respectively positioned at two sides of the gold film. The included angle between the incident light and the normal line of the gold film is an incident angle, and the rotating platform is rotated to continuously change the incident angle within the range of 0-90 degrees. The focus of the fluorescence collecting lens is located on the surface of the gold film or within a defocus range from one particle diameter of the metal film.
Claims (9)
1. A fluorescence detection method of unmarked micro-nano particles is characterized in that,
exciting surface plasma resonance of noble metal by using a total internal reflection structure, generating restrained total internal reflection after the micro-nano particles positioned on the surface of the noble metal film and the surface plasma resonance of the noble metal film resonate, exciting a fluorescent solution where the micro-nano particles are positioned to emit light by transmitted light of the micro-nano particles, wherein the untreated micro-nano particles and the fluorescent solution are injected into a sample groove; the fluorescence emitted from the sample groove is collected by the fluorescence collecting lens and converged to the photoelectric detector after passing through the optical neutral filter.
2. The fluorescence detection method according to claim 1, wherein the fluorescence detection method,
the device includes: the device comprises a laser (1), a polarization regulator (2), a light wave vector-metal film surface plasma coupler (3), a glass slide glass with one side surface plated with a noble metal film (4), a bottom-lacking sample groove (5), a fluorescence collecting lens (6), an optical neutral filter (7), a photoelectric detector (8), a processing circuit (9), a computer (10) and a rotary table (11);
wherein the laser (1) provides incident light for exciting surface plasmon resonance of the noble metal film (4); the polarization regulator (2) regulates the polarization state of incident light emitted by the laser (1) into p light parallel to an incident plane, and the p light is incident on the noble metal film of the glass slide through the light wave vector-metal film surface plasma coupler (3), and the coupler (3) and the glass surface below the noble metal film (4) in the glass slide are connected together through a refractive index matching substance; the bottom-lacking sample groove (5) is directly placed on the noble metal film and sealed with the noble metal film to form a sample groove, and untreated micro-nano particles and fluorescent solution are injected into the sample groove; fluorescence emitted from the sample cell is collected by a fluorescence collecting lens (6), converged to a photoelectric detector (8) after passing through an optical neutral filter (7), and input into a computer (10) after passing through a processing circuit (9); the optical neutral filter (7) is used for setting the threshold value of the fluorescence entering the photoelectric detector; the light wave vector-metal film surface plasma coupler (3) is used for matching light wave vectors with wave vectors of the noble metal film; the light wave vector-metal film surface plasma coupler (3) is connected with the glass bottom of the sample groove plated with the noble metal film through a refractive index matching material and fixed on the rotating table (11); the computer (10) controls the rotating table to rotate continuously.
3. The device according to claim 2, wherein the noble metal film has a thickness h in the range of 35nm ≦ h ≦ 75 nm.
4. The device according to claim 2 or 3, wherein the excitation light excites the surface plasmon resonance of the noble metal film (4) through the light wave vector-metal film surface plasmon coupler (3), and the excitation light and the sample cell containing the micro-nano particles to be detected are respectively positioned at two sides of the noble metal film.
5. The device of claim 4, wherein the wavelength of the incident light is within the full width at half maximum of the absorption peak of the fluorescent solution, and the bandwidth of the incident light is determined by the diameter and concentration of the particles to be measured.
6. The device according to claim 2, 3 or 5, wherein the diameter range of the micro-nano particles to be detected is 30nm-1000nm, and the surface of the particles is made of non-metallic materials.
7. The device according to claim 2, 3 or 5, wherein the angle between the incident light and the normal of the noble metal film is an incident angle, and the incident angle is continuously changed within a range of 0 to 90 degrees by rotating the turntable.
8. The device according to claim 2, 3 or 5, wherein the focus of the fluorescence collecting lens is located on the surface of the noble metal film or within a defocus range from one particle diameter of the noble metal film.
9. A device according to claim 2, 3 or 5, wherein the light source is selected to be one of: 1) single bundle; 2) a plurality of identical light sources having the same incident angle; 3) and multiple beams with the same incident angle are separated from the same light source.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710467212.4A CN107290314B (en) | 2017-06-20 | 2017-06-20 | Fluorescence detection method and device for unmarked micro-nano particles |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710467212.4A CN107290314B (en) | 2017-06-20 | 2017-06-20 | Fluorescence detection method and device for unmarked micro-nano particles |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107290314A CN107290314A (en) | 2017-10-24 |
| CN107290314B true CN107290314B (en) | 2020-10-20 |
Family
ID=60097895
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710467212.4A Active CN107290314B (en) | 2017-06-20 | 2017-06-20 | Fluorescence detection method and device for unmarked micro-nano particles |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107290314B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108593416B (en) * | 2018-04-08 | 2020-09-08 | 国家纳米科学中心 | Micro-nano particle detection system and method |
| CN111141715B (en) * | 2020-02-27 | 2022-09-20 | 南京理工大学 | Device and method for measuring three-dimensional orientation of fluorescent molecules based on rotating orthogonal slits |
| CN112033913B (en) * | 2020-08-25 | 2024-03-22 | 中国科学院合肥物质科学研究院 | Device and method for measuring water content of nano single particles based on surface plasmon resonance imaging |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101074921A (en) * | 2006-05-18 | 2007-11-21 | 中国科学院化学研究所 | Multifunctional light-absorbing, scattering and transmitting spectrograph based on surface plasma wave |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010091553A (en) * | 2008-09-09 | 2010-04-22 | Konica Minolta Holdings Inc | Method for detecting biomolecule |
| US9315855B2 (en) * | 2011-02-16 | 2016-04-19 | Centrul International De Biodinamica | Systems and methods for detection and quantitation of analytes using an oscillating stimulus |
-
2017
- 2017-06-20 CN CN201710467212.4A patent/CN107290314B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101074921A (en) * | 2006-05-18 | 2007-11-21 | 中国科学院化学研究所 | Multifunctional light-absorbing, scattering and transmitting spectrograph based on surface plasma wave |
Non-Patent Citations (3)
| Title |
|---|
| Surface plasmon resonance scattered by a dielectric sphere;Xin Hong 等;《Proc. of SPIE》;20161103;第10028卷;1-6 * |
| Surface Plasmon resonance study of vesicle rupture by virus-mimetic attack;Soonwoo Chah 等;《Physical Chemistry Chemical Physics》;20080421;第10卷;3203-3208 * |
| Xin Hong 等.Surface plasmon resonance scattered by a dielectric sphere.《Proc. of SPIE》.2016,第10028卷 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107290314A (en) | 2017-10-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Welsh et al. | A compendium of single extracellular vesicle flow cytometry | |
| CN107290314B (en) | Fluorescence detection method and device for unmarked micro-nano particles | |
| JPH0544621B2 (en) | ||
| JP2014517322A5 (en) | ||
| US9709476B2 (en) | System and method for finding the size of one of more individual particles, such as nanoparticles for example, in real time utilizing a microsphere's whispering gallery modes (“WGMs”) | |
| US7511820B2 (en) | Surface plasmon resonance sensing system and method thereof | |
| US20130260479A1 (en) | Device and method for detecting existence of target biomolecules in a specimen | |
| US10203283B2 (en) | Fluorescent substance detection system | |
| CN104964964A (en) | Portable laser raman spectrometer based on prismatic decomposition | |
| Baquedano et al. | Low-cost and large-size nanoplasmonic sensor based on Fano resonances with fast response and high sensitivity | |
| CN106568747A (en) | Optical waveguide fluorescence enhanced detector | |
| Berg et al. | Two-dimensional Guinier analysis: application to single aerosol particles in-flight | |
| JP2009063462A (en) | Optical measuring instrument and particulate analyzer | |
| CN210923475U (en) | Serum albumin detection system based on optical fiber SPR sensor | |
| CN204028004U (en) | A kind of substance detecting apparatus based on Raman filtering | |
| Wu et al. | Development of a Low-Cost, Sensitivity-Optimized Fluorescence Sensor for Visible Spectrum Analysis | |
| EP2227685B1 (en) | Microelectronic sensor device | |
| CN104048915A (en) | Real-time monitoring device and method of optical material and laser interaction process | |
| CN111965161A (en) | Optical fiber surface enhanced Raman spectrum sensing detection device and detection method | |
| CN110261964B (en) | Optical fiber head for optical fiber spectrometer | |
| CN104236604A (en) | Fluorescence method optical fiber sensor | |
| US10495575B2 (en) | Surface plasmon enhanced fluorescence measurement device and surface plasmon enhanced fluorescence measurement method | |
| CN108414480B (en) | Crude oil fluorescence measuring device and method | |
| Sada et al. | Heavy metal detection based on coreless fibers using the LSPR technique | |
| CN117538135B (en) | Aerosol LIBS detection composite film based on nano structure and detection method |
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 |