CN103698510A

CN103698510A - Analyzing chemical and biological substances using nano-structure based spectral sensing

Info

Publication number: CN103698510A
Application number: CN201310688128.7A
Authority: CN
Inventors: 汪泓; 郭浔; 刘春伟; 邓安平; 常化仿
Original assignee: Opto Trace Technologies Inc
Current assignee: Opto Trace Technologies Inc
Priority date: 2012-12-21
Filing date: 2013-12-16
Publication date: 2014-04-02

Abstract

An integrated chromatography-immunoassay system for integrated chromatography-immunoassay system includes a chromatographic unit that receives labeled nano-structured probes comprising nano particles and antibodies attached to the nano particles, and a test membrane comprising coating antigens. The chromatographic unit allows the labeled nano-structured probes to diffuse there through and into the test membrane, wherein the antibodies on the nano particles are bound to the coating antigens. A laser device emits a laser light to illuminate the labeled nano-structured probes having the antibodies bound to the coating antigens on the test membrane. A spectral analyzer obtains a Raman spectrum from light scattered from the labeled nano-structured probes having the antibodies bound to the coating antigens on the test membrane, and to identify a spectral signature in the Raman spectrum associated with the antibody-antigen pair, which enables detection and identification of the antibody.

Description

Use spectrum detection method analytical chemistry thing based on nanostructured and the method for biological

The application is the common unsettled U.S. Patent application US13/442 transferring the possession of, 835 continuity application also requires its right of priority, U.S. US13/442, the title of No. 835 patented claims is " using the spectrum detection method based on nanostructured to guarantee food security ", the applying date is on April 9th, 2012.U.S. US13/442, No. 835 patented claims require U.S. Provisional Application US61/507,592 right of priority, U.S. US13/442, the title of No. 835 patented claims is " using spectrum detection method analytical chemistry thing and biological based on nanostructured ", and the applying date is on July 13rd, 2011.At this, disclose these related application and be incorporated to by reference the application.

Technical field

The present invention relates to material detection field, relate in particular to a kind of method of utilizing light scattering probe and chemical detector to detect chemical substance, biochemical substances, radiomaterial and other materials.

Background technology

Light scattering technique for example Raman spectroscopy can detect chemical substance and biochemical substances.A major limitation of application Raman spectroscopy is, the Raman scattering signal obtaining from chemical substance and biochemical substances very a little less than.Although people are doing many effort aspect enhancing Raman scattering intensity, but still there is no to produce the practical and economic detecting device based on Raman spectroscopy.Therefore, the application of the detection chemical substance of Raman scattering method up to now and biochemical substances aspect is also very limited.

So, be necessary to provide a kind of effective and practical detecting device based on Raman spectroscopy to detect and to survey micro-trace chemistry material, biochemical substances, radiomaterial and other materials.

Summary of the invention

The application discloses a kind of Raman spectrum detection (or detection) system and method that has supersensitive detection and survey chemical substance or biochemical substances.The biochemical substances that Raman spectrum detection system disclosed by the invention and method can detect or detect or chemical substance concentration level are than the low several orders of magnitude of conventional detection technique.Biochemical substances and chemical substance comprise antigen, antibody and widely little molecule and metallic element.Particularly, when being applied to biochemical immunity mensuration, detection sensitivity and specificity can obviously improve.

On the one hand, the present invention relates to a kind of integrated immunochromatography system, comprise a chromatography unit, can receive marking nano structure probe, this marking nano structure probe comprises nano particle, the detection film that is adsorbed on the antibody on nano particle and contains envelope antigen, wherein chromatography unit allows marking nano structure probe to diffuse through and enter detection film, and wherein the antibody on nano particle is incorporated on envelope antigen.Integrated immunochromatography system also comprises a laser aid, can Emission Lasers with exposure label(l)ing nanostructure probe, this marking nano structure probe has the antibody being combined on the envelope antigen detecting on film, an and spectroanalysis instrument, can obtain the Raman spectrum from the scattered light of marking nano structure probe, this marking nano structure probe has the antibody being combined on the envelope antigen detecting on film, and identification with antibody-antigen to the spectral signal in the Raman spectrum being associated, thereby realize the detection and Identification of antibody.

The enforcement of said system can comprise following one or more.Chromatography unit can receive competitive antigen and allow competitive antigen to diffuse through and enter detection film, and wherein competitive antigen and envelope antigen are competed to be attached on the antibody of nano grain surface.Marking nano structure probe can comprise the spectrum mark being combined on nano particle or antibody.Antigen can include but not limited to Clenbuterol (clenbuterol), nitrofuran (nitrofurans), organophosphorus (organic phosphorus), organochlorine (organochlorine), positive terephthalate (pro terephthalate), pesticide (a pesticide), rat-bane (a rodenticide) or veterinary drug (a veterinary drug).Antigen can comprise melamine (melamine), honey element (sodium cyclamate), sodium cyclohexylsulfamate (sodium cyclohexylsulfamate), sucrose (cane sugar), starch (starch), the Sudan I, II, III and IV(Sudan I, II, III and IV), malachite green (malachite green), acephatemet (methomidophos), orthene (acephate), DDT(dichloro-diphenyl-trichloroethane), DDV(DDVP), malathion (malathion), fenifrothion (fenitrothion), decis (deltamethrin), cypermethrin (cypermethrin), parathion-methyl (methyl parathion), phosmet (phosmet), Rogor (dimethoate), nitrofuran (nitrofuran), furazolidone (furanzolidole), chloromycetin (chloramphenicol), duomycin (chlortetracycline), Ciprofloxacin (ciprofloxacin), Clenbuterol or Enrofloxacin (enorfloxacin).The mean diameter of nano particle is within the scope of 10-100nm.The material that nano particle can comprise is selected from metal, metal alloy, oxide, silicon, composite materials, magnetic or ferrimagnet and their combination, or is selected from Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and their combination.Envelope antigen can comprise Clenbuterol, and wherein Clenbuterol is incorporated on chicken ovalbumin (OVA) molecule, forms OVA-Clenbuterol complex, and this OVA-Clenbuterol complex is attached to and detects on film.

On the other hand, the present invention relates to a kind of substance identification.The method comprises is adsorbed onto in Nanosurface structure antibody, introduce unknown antigen to Nanosurface structure, so that unknown antigen combines with the structural antibody of Nanosurface, utilize the unknown antigen and the antibody that in laser beam irradiation Nanosurface structure, combine, the unknown antigen combining in collection Nanosurface structure and the scattered light of antibody, acquisition is from the Raman spectrum of the unknown antigen combining in Nanosurface structure and antibody scattered light, find the spectral signal being associated with the unknown antigen combining and antibody in Raman spectrum, and the unknown antigen matching according to Raman signal identification and the structural antibody of Nanosurface.

The enforcement of said system can comprise following one or more.Antigen can be selected from material (a substance in a veterinary drug), arsenic compound (an arsenic compound) and the prussiate (a cyanide) in Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane, veterinary drug.Antigen also can be selected from melamine, honey element, sodium cyclohexylsulfamate, sucrose, starch, the Sudan I, II, III and IV, malachite green, acephatemet, orthene, DDT, DDV, malathion, fenifrothion, decis, cypermethrin, parathion-methyl, phosmet, Rogor, nitrofuran, furazolidone, chloromycetin, duomycin, Ciprofloxacin, clenbuterol hydrochloride or Enrofloxacin.The method further comprises the identifying and diagnosing human body diseases of the unknown antigen based on disease association.Reactant can comprise the body fluid obtaining from human body.Disease can include but not limited to cancer, asthma, allergy, cirrhosis, renal failure, leukaemia, Alzheimer's, Parkinson's disease, diabetes, craving for tobacco disease, arthritis, cardiovascular disease, serious acute respiratory syndrome (SARS), influenza and acquired immune deficiency syndrome (AIDS) (HIV).Unknown antigen can comprise forbidden drug, includes but not limited to heroin (heroin), dexoxyn (methamphetamine), cocaine (cocaine), caffeine (caffeine), morphine (morphine), codeine (codeine), amphetamine (amphetamine), ephedrine (ephedrine), papaverine (papaverine), narcotine (narcotine), KET (Ketamine) and head-shaking pill (MDMA).Unknown antigen can comprise protein carrier and be attached to the unknown materials molecule on protein carrier, and wherein the identification of protein carrier causes the identification of unknown materials.Unknown materials can comprise KET or Fluorakil 100 (fluoroacetamide).Unknown antigen can be extracted from food.

When the method is further included in the scattered light that gathers Nanosurface structure, apply electric field, magnetic field or electromagnetic field to Nanosurface structure.Nanostructured surface can comprise the nano particle of mean diameter within the scope of 1-1000nm.The material that nano particle comprises is selected from metal, metal alloy, oxide, silicon, composite materials, magnetic or ferrimagnet and their combination, or is selected from Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and their combination.Nanosurface structure can form in substrate.Nanosurface structure is included in projection or the column that upper surface of substrate forms, and wherein adjacent projection or column mean diameter are between 10-1000nm.Nanosurface structure can be included in depression or the hole that upper surface of substrate forms.The mean diameter of adjacent recessed or hole is between 10-1000nm.

On the other hand, the present invention relates to a kind of substance identification.The method comprises pretreated chemical substance or biochemical substances is applied to Nanosurface structure; The reactant that introducing comprises unknown materials, to Nanosurface structure, makes the molecule of unknown materials under the assistance of pretreated chemical substance or biochemical substances, be adsorbed onto Nanosurface structure; Utilize laser beam irradiation absorption to have the Nanosurface structure of the molecule of unknown materials; Collection is had the scattered light of Nanosurface structure of the molecule of unknown materials by absorption; From adsorbing the scattered light of the Nanosurface structure of the molecule that has unknown materials, obtain Raman spectrum; Find the spectral signal being associated with unknown materials in Raman spectrum; And utilize the unknown materials in Spectral signal pattern recognition reactant.

The enforcement of said system can comprise following one or more.Antigen can comprise protein carrier and be combined in the molecule of the unknown materials on protein carrier.The method further comprises according to Spectral signal pattern recognition protein carrier, and wherein the identification of unknown materials is to carry out according to the identification of protein carrier.Unknown materials can comprise KET or Fluorakil 100.Pretreated chemical substance or biochemical substances can comprise the antibody that can be adsorbed onto on Nanosurface body structure surface, and wherein unknown materials is the antigen matching with antibody, wherein Raman signal be adsorbed onto the structural Ag-Ab of Nanosurface to being associated.Pretreated chemical substance or biochemical substances can comprise the antigen that can be adsorbed onto on Nanosurface body structure surface, and wherein unknown materials is the antibody matching with antigen, wherein spectral signal be adsorbed onto the structural Ag-Ab of Nanosurface to being associated.Nanosurface structure can comprise the nano particle of mean diameter within the scope of 1-1000nm, or the nanostructured forming in substrate.

Accompanying drawing explanation

Following accompanying drawing is as a part for instructions, and diagram embodiments of the invention, are used for illustrating principle of the present invention together with summary of the invention.

Figure 1A-1C for example understands the system of utilizing a Raman scattering probe to detect chemical substance and biochemical substances.

Fig. 2 is the schematic diagram of an example probe that is applicable to the Raman scattering probe of Figure 1A-1C.

Fig. 3 A and 3B are respectively the schematic diagram that uses Raman probe to check passenger and luggage on airport.

Fig. 4 A is the schematic diagram for the Raman scattering probe wired connection network of monitoring building safety.

Fig. 4 B is the schematic diagram for the Raman scattering probe wireless connections network of monitoring building safety.

Fig. 4 C Shi Nengyu control center carries out an example structure figure of the probe assembly of radio communication.

Fig. 4 D is the time of hazard recognition substance source and an example flow diagram of position.

Fig. 5 is the schematic diagram that uses Raman scattering probe to carry out environmental monitoring.

Fig. 6 A is used Raman probe to check the schematic diagram of food security.

Fig. 6 B is used Raman scattering probe to carry out the schematic diagram of long-range medical diagnosis on disease and the biomedical system detecting.

Fig. 6 C is the schematic diagram that uses a plurality of Raman scattering probes to carry out quality of production control in a multi-channel detection system.

Fig. 6 D is used a plurality of Raman scattering probes to carry out the schematic diagram of safety and quality check and the drug identification of counterfeit goods detection, Food & Drink in a multi-channel detection system.

Fig. 7 utilizes the solution that contains nano particle and light scattering probe to detect the schematic diagram of trace chemistry thing or biological substance according to one embodiment of present invention.

Fig. 8 is the process flow diagram that utilizes the solution contain nano particle and light scattering probe to detect trace chemistry thing or biological substance.

Fig. 9 A is the micro-image example of using the nano particle that sem observation arrives.

Fig. 9 B is the particle diameter distribution example figure of the nano particle in solution shown in Fig. 7.

Figure 10 is the cut-open view for the manufacture of the multilayer layer structure of nanostructured.

Figure 11 A forms the cut-open view of hole in the multilayer layer structure shown in Figure 10.

Figure 11 B is the top view of the multilayer layer structure shown in Figure 11 A.

Figure 11 C is that multilayer layer structure shown in Figure 11 B is along the cut-open view of A-A line.

Figure 12 is at the cut-open view of the structrural build up nanostructured of multilayer stratiform after wet chemical etching technique or chemically mechanical polishing.

Figure 13 is at the cut-open view of the structrural build up nanostructured of multilayer stratiform after removing the restraining barrier of hole bottom and being etched to conductive layer.

Figure 14 A is at the cut-open view of the structrural build up nanostructured of multilayer stratiform after depositing noble metal.

Figure 14 B is the cut-open view of removing after the noble metal of top layer in the structrural build up nanostructured of multilayer stratiform.

Figure 15 is the cut-open view of removing after oxide layer in the structrural build up nanostructured of multilayer stratiform.

Figure 16 A-16D, 16G and 16H are at the cut-open view of the structrural build up nanostructured of multilayer stratiform after corresponding manufacture craft.

Figure 16 E and 16F are at the top view of the structrural build up nanostructured of multilayer stratiform after corresponding manufacture craft.

Figure 17 detects the exemplary plot of the raman spectral signal of carcinoma of mouth by Raman scattering probe disclosed by the invention in an oral cavity carninomatosis people saliva.

Figure 18 detects the exemplary plot of the raman spectral signal of breast cancer by Raman scattering probe disclosed by the invention in the saliva of a Breast Cancer Patients.

Figure 19 A and 19B are used Raman scattering probe disclosed by the invention the exemplary plot of the raman spectral signal of lung cancer in the saliva of a lung cancer patient and serum, to be detected.

Figure 20 detects the exemplary plot of oophoroma raman spectral signal by Raman scattering probe disclosed by the invention in an oophoroma patient serum.

Figure 21 detects the exemplary plot of acquired immune deficiency syndrome (AIDS) raman spectral signal by Raman scattering probe disclosed by the invention in the saliva of an AIDS patients.

Figure 22 detects the exemplary plot of the raman spectral signal that uses forbidden drug in a forbidden drug user saliva by Raman scattering probe disclosed by the invention.

Figure 23 example has illustrated by Raman scattering disclosed by the invention and has popped one's head in, through with N-first-2-5-pyrrolidone (can terraced Buddhist nun, the metabolic product of nicotine) raman spectral signal is made comparisons, and the raman spectral signal that can determine smoking state detected in a smoker saliva.

Figure 24 is the process flow diagram that uses Raman scattering probe disclosed by the invention to carry out non-intrusion type medical diagnosis on disease.

Figure 25 is the process flow diagram that the immunoassays of the spectral detection based on nanostructured used according to the invention detect unknown antigen process.

Figure 26 A example has illustrated as described in Figure 25 utilizes the spectral detection based on nano particle to carry out the preparation of the sample of immunoassays in process.

Figure 26 B example has illustrated the spectral detection of the sample of the nano particle that comes self-contained absorption to have antibody in process as described in Figure 25.

Figure 26 C has shown the introducing of unknown antigen to be measured in the sample of nano particle that comprises absorption in process as described in Figure 25 and have antibody.

Figure 26 D has shown that as described in Figure 25 in process, unknown antigen is attached to the detection on the antibody adsorbing on nano particle.

Figure 27 is the process flow diagram that the immunoassay of the spectral detection based on nanostructured used according to the invention detects another process of unknown antigen.

Figure 28 A example has illustrated to be prepared to utilize spectral detection based on nanostructured to carry out the structure of the sample of immunoassays for process as described in Figure 27.

Figure 28 B example has illustrated the spectral detection configuration of the sample of the nanostructured of coming self-contained absorption to have antibody in process as described in Figure 27.

Figure 28 C has shown the detection that is attached to the unknown antigen on the antibody that nano particle adsorbs in process as described in Figure 27.

Figure 29 has shown that the spectral detection based on nanostructured used according to the invention carries out the competitive assay process of the micromolecular compound of immunoassays.

Figure 30 A-31B example has illustrated the integrated immunochromatography system of the light-scattering material detection being provided for based on nanostructured.

Figure 32 example has illustrated the mark of nanostructure probe, example antigen and envelope antigen that Figure 30 A-31B is used.

Figure 33 example has illustrated the preparation that is applicable to the nanostructure probe of integrated immunochromatography system.

Figure 34 has shown the detail view for the scattered light of the nanostructure probe of integrated immunochromatography system.

Figure 35 example has illustrated the implementation step of using the light scattering based on nanostructured that the integrated immunochromatography system shown in Figure 30 A-31B carries out and material to detect.

Figure 36 has shown the Raman scattering intensity as competitive antigen concentration function in integrated immunochromatography system.

Figure 37 has shown the standard Raman scattering curve of integrated immunochromatography system.

Figure 38 has shown the detection line photo in the detection film being obtained by integrated immunochromatography system.

Figure 39 example has illustrated the raman scattering spectrum under different preparatory conditions.

Figure 40 has shown the repeatability of the Raman scattering intensity peak being obtained along middle 10 the different points of detection line by integrated immunochromatography system.

Figure 41 has shown the specificity of integrated immunochromatography system.

Embodiment

Figure 1A-1C has illustrated respectively use Surface enhanced raman spectroscopy (Surface-Enhanced Raman Scattering, SERS, or Nano-Enhanced Raman Scattering, NERS) to detect or survey the system of micro-trace chemistry material or biochemical substances.Referring to Figure 1A, a light scattering probe 100 comprises that a probe 110 and is positioned at the other detecting device 105 of probe 110.Detecting device 105 comprises a Nanosurface structure.For example, this Nanosurface structure can comprise shown in a plurality of nano-pillar 108(Figure 1B), a plurality of nanoaperture, the nano particle bunch in solution, or the surface structure of other nano-scale.In certain embodiments, as described below, can make Nanosurface structure by the colloidal suspension solution that contains nano particle being coated in to the surface of detecting device 105.Then solution can be evaporated, and nanoparticle deposition is to surface.In instructions of the present invention, term " nano particle " refers to the granule that is at least less than 1,000nm in the size of a direction.

In certain embodiments, sample solution can be guided in the nano-pillar 108 of detecting device 105.Described sample solution can comprise patient or forbidden drug user's body fluid, to diagnose the illness and definite drug use situation, or the fluid of gas phase.The example of body fluid includes but not limited to blood, saliva, urine, serum, tear, sweat, gastric juice, hydrothorax, ascites, celiolymph (CSF), seminal fluid and juice.Sample solution can also comprise food samples, for guaranteeing harmful in food safety detection food samples or illegal additive.The example of food includes but not limited to dairy products, for example liquid milk, milk powder, babies ' formula milk powder, cheese, Yoghourt, ice cream, and other are containing milk deli, and for example, containing candy, cake and the biscuit of milk, and proteinaceous food.Probe 110 and detecting device 105 can be to pack in a probe assembly 120.Probe assembly 120 can reduce pressure to reduce the pollution of impurity to sensitive surface by a vacuum pump.

The laser beam of laser instrument 141 transmitting is transmitted to probe through optical fiber 125, is radiated at detecting device 105(Figure 1A) Nanosurface structure on, as shown in Figure 1 C.The scattered light that sample solution on detecting device 105 Nanosurfaces produces gathers by probe 110, through optical fiber 130, is transmitted to spectrometer 140.The output signal of spectrometer 140 obtains the Raman spectrum of scattered light by spectroanalysis instrument 150.Identify the one or more spectral signals in Raman spectrum, and the spectral signal of itself and predetermined different kinds of molecules is made comparisons.When detect finding to have exceeded the threshold value of certain molecule, output signal 160 Identification displays are to target molecule, for example, and the objectionable impurities in food or water, dangerous substance (cruelly exploding property or inflammability material), or disease association molecule.In instructions of the present invention, term " spectral signal " refers to one or more spectrum peaks, one or more spectrum paddy, and other wave spectrum shape, for example relatively peak height, peak line width, peak shape etc., they have characterized the one or more molecular links in biochemical, medicine or chemical substance.

In certain embodiments, detecting device 105 can comprise nanostructureds different on detecting device, as the unsettled U.S. Patent application US12/014 at common transfer, 800(authorizes as US Patent No. 7 on January 11st, 2011,869,044) disclosed in, its title is " Systems for optical inspection on micro array structure ", the applying date is on January 16th, 2008, and the content of above-mentioned application is incorporated to the application by reference.Above-mentioned Systems for optical inspection comprises a fluorescence detector, described fluorescence detector comprises a substrate, this substrate has upper surface and is positioned at a plurality of conical wall on substrate, wherein at least one conical wall is longitudinally arranged, wherein a plurality of conical wall comprise the skewed surface that is pitch angle with respect to upper surface, and wherein skewed surface is used to adsorb chemical example molecule; One light source is used for launching incident beam a plurality of conical wall of chemical example molecule to incide absorption; And one the collector scattered light that is used for gathering a plurality of conical wall to determine chemical example.

Detecting device 105 also can comprise that one has substrate and suprabasil a plurality of conical wall of upper surface, wherein a plurality of conical wall comprise the skewed surface that is pitch angle with respect to upper surface of substrate, wherein at least two adjacent conical wall define an air gap, and the width that this air gap has is the function with upper surface distance; One light source, being used to launch incident beam has the conical wall of chemical example molecule to incide absorption; And a collector, the scattered light that is used for gathering a plurality of conical wall is to determine chemical example.

In certain embodiments, detecting device 105 can comprise multi-layer nano structure, as the unsettled U.S. Patent application US11/754 of common transfer, 912(authorizes as US Patent No. 7 on February 22nd, 2011,892,489) disclosed multi-layer nano post and multi-layer nano hole in, the name of this application is called " light scattering device with multiplelayer microstructure ", the applying date is on May 29th, 2007, nano particle bunch or group, or the multi-layer nano particle in solution, the content of above-mentioned application is incorporated to the application by reference.Nanosurface structure can comprise silicon base; Be positioned at the adhesion layer in silicon base; Be positioned at the bias layer on adhesion layer; With the one or more structural sheets that are positioned on adhesion layer.One or more structural sheets can comprise different material formations and pass at least two-layer a plurality of holes in two or more structural sheets.The width in a plurality of holes is at 0.5-1, within the scope of 000nm.Nanosurface structure also can comprise a silicon base; One is positioned at the adhesion layer in silicon base; One is positioned at the bias layer on adhesion layer; With a plurality of posts that are positioned on bias layer.At least one in a plurality of posts or hole can comprise have that different material forms and width at 0.5-1, the two or more structural sheets within the scope of 000nm.

Referring to Fig. 2, the laser beam that probe 110 receives from input optical fibre 125.This laser beam projects on detecting device 105 by a bandpass filter 170, a lens combination 175-1 and 175-2 successively.From detecting device 105 scattered light out, through a catoptron group 180-1 and 180-2, be guided through another bandpass filter 185, further through collimation lens 190, enter and collect optical fiber 130.

Detected micro-trace chemistry or biochemical preparation can be gas, liquid, solid, collosol and gel or aerocolloidal form.Its molecule is adsorbed on the Nanosurface or nano particle of detecting device 105.Compare with the molecule being freely present in gas, liquid, solid, collosol and gel or gasoloid, these molecules that are attracted on Nanosurface or nano particle have much bigger scattering cross-section under the irradiation of laser beam.When minute period of the day from 11 p.m. to 1 a.m that these are adsorbed with laser beam irradiation, can obtain the raman scattering spectrum of these molecules.The predetermined raman spectral signal comparison that is stored in system database by these molecules can identify target chemistry or biochemical preparation.

Fig. 3 A has shown an example that strengthens Raman scattering in traffic safety field application surface.Passenger 200-1,200-2 and the 200-3 by walkway carried out to examination.One or more probe assemblies 120 with embed-type detecting device 105 are installed in walkway 210.Near spectroanalysis instrument 150 probe assembly 120 can connect by optical fiber or in the office of distant place.In probe assembly 120, probe and detecting device (or detector) are packaged together.The sensitive surface of probe direct detection device 105.Walkway 210 can be artificial draft and at slight negative pressure and/or slightly high temperature, to strengthen the volatilization of objectionable impurities or to transport.For example, if a passenger (passenger 200-2) carries explosive, harmful chemical, chemical weapons, flammable liquid, chemical and biological weapons, nuclear weapon or anaesthetic, these materials of microscratch amount are by volatilization or transport and enter in air, by custom-designed sample collection system (U.S. US7,384, No. 792 patent has disclosed relevant details), these molecules are adsorbed to the surface of detecting device.Record Raman spectrum and the spectral signal of the known substance storing in itself and central office database is compared.Once objectionable impurities be detected, just trigger alarm, take suitable safety practice.

Referring to Fig. 3 B, goods 215 transports by goods examination passage 220 via a travelling belt 230.Embedding have the probe assembly 120 of detecting device 105 to be placed in goods examination passage 220 everywhere.The spectroanalysis instrument 150 of probe assembly 120 in optical fiber coupling or distant place office.Probe assembly 120 is aimed at the surface of detecting device 105, and packaging together with detecting device 105, for detection of any explosive, chemistry or chemical and biological weapons or Harmful chemicals being contained in goods 215.This facility can be applicable to the various places that need to check passenger baggage, for example airport, post office, railway station, subway station, customs inspection post, traffic control area, steamer or submarine, aircraft, school, hotel, restaurant, shopping center, recreation center, buildings and other public places, etc.With this facility, be easy to implement to detect gunpowder and other explosives, the container that contains liquid inflammable and explosive substances or other dangerous goods.

wired detector network

Referring to Fig. 4 A, one detects (or detection) device network system 400 is applied to the safety monitoring of public building, and described public building is airport, railway, bus stop, stadiums, government bodies, auditorium, cinema, law court, shopping mall, other public buildings or public gathering occasion for example.A plurality of probe assemblies 401 are installed on the diverse location in public building 415 or other defence region.Each probe assembly 401 comprises a probe and a detecting device (not showing respectively in Fig. 4 A).This probe can with probe 110(Figure 1A) similar, but can also comprise a laser aid.This detecting device is consistent with the detecting device 105 shown in Figure 1B, comprises its lip-deep Nanosurface structure.This detecting device can also comprise the liquid solution detecting in order to receive reactant.This solution can comprise the nano particle of adsorption reaction thing molecule.Probe assembly 401, for monitoring various material molecule, provides early detection for entering into the chemicals any danger or harmful of guarded region.The light signal that probe assembly 401 gathers is input to the optical multiplexer 403 in control roller office 408 through optical fiber 402 hyperchannels.Light signal decomposes and produces spectral signal through spectrometer 404, and spectral signal is analyzed by spectroanalysis instrument 405 again.Processor 407 utilizes the spectral information being pre-stored in database 406 to identify spectral signal in spectroscopic data.Some special cases of supervision on hazardous substances include but not limited to explosive detection, comprise liquid explosive, chemistry or chemical and biological weapons (comprising anthrax), flammable liquid material, drugs etc.

wireless detector network

In certain embodiments, detector network system 430 as shown in Figure 4 B.Buildings 415 comprises an entrance and multiaspect wall.Probe assembly 410A-410I is arranged on the diverse location of buildings 415.Probe assembly 410A-410I connects together with antenna 411A-411I respectively, and the spectral information that antenna 411A-411I detects locality is transmitted into control center 450.

As shown in Figure 4 C, each probe assembly 410A comprises that a probe 420A and is positioned at the detecting device 430A of probe 420A adjacent position.Detecting device 430A can gather the material in surrounding environment.In certain embodiments, detecting device 430A comprises a nanostructured surface, is adsorbed on the molecule of the material collecting in surrounding environment.The laser instrument 421A(that probe 420A comprises a compactness is semiconductor laser instrument for example), laser instrument 421A is to being adsorbed in the sample molecule illuminating laser beam of detecting device 430A.Probe 420A further comprises optical signalling harvester 422A, and its collection is adsorbed in the light signal of the sample molecule scattering of detecting device 430A, the correlation molecule information that these scattering envelopes contain sample molecule.Detecting device 430A and detecting device 105(Figure 1B) consistent, on its surface, there is a nanostructured.Be adsorbed in the structural sample molecule scattering of this Nanosurface incident laser.Detecting device 430A can also comprise that one for receiving the liquid solution (referring to Fig. 7 below) of reactant to be measured.This solution can also comprise the nano particle for adsorption reaction thing molecule.

Probe 420A also comprises the spectrometer 440A of a compactness, and the scattered light that probe 420A collects produces a spectrum through spectrometer 440A.From the spectroscopic data of spectrometer 440A output, be transferred to wireless communication line 445A.Wireless communication line 445A can comprise a radio frequency (RF) transceiver, one or more amplifiers and impedance matching circuit.Wireless communication line 445A is transferred to the 450(Fig. 4 B of control center for the spectroscopic data that probe assembly 410A is detected).

Referring to Fig. 4 B, control center 450 comprises a wireless router 460 with antenna 455, for receiving the wireless signal from antenna 411A-411I, and from this wireless signal, produces the electric signal that comprises spectroscopic data.Control center 450 can be from signal source (for example, buildings 415) close together (for example, within several miles), allow the wireless signal that comprises spectroscopic data transmit according to a home control network communication protocol, for example bluetooth, WiMax, WiBro, WiFi, WLAN, 802.16 and other home control network communication protocol.Control center 450 can also from signal source away from, the wireless signal that wherein comprises spectroscopic data can utilize wireless communication standard and agreement to transmit, for example gsm (GSM), universal mobile telecommunications service (UMTS) and CDMA (CDMA).GSM can comprise GPRS EDGE and CSD.UMTS can comprise Wideband Code Division Multiple Access (WCDMA) (WCDMA), high-speed packet access (HSPA), high-speed downlink packet access (HSDPA), universal mobile telecommunications system time division duplex (UMTS-TDD) and Long Term Evolution (LTE) technology.CDMA can comprise CDMA2000 and Ultra-Mobile Broadband (UMB).

The spectroanalysis instrument 465 of control center 450 is for receiving the electronic signal that comprises spectroscopic data from wireless router 460.Analysis by spectroanalysis instrument 465 obtains a spectrum (for example Raman spectrum).As described below, different chemical substances or biochemical substances often have unique spectral signal.These spectral signals can utilize known chemical substance or a biochemical substances, and one is similar to the detecting device being arranged in probe assembly 410A-410I and measures in advance.These spectral signals can be kept in database 470.Spectroanalysis instrument 465 can use the spectral signal being kept in database 470 spectral signal identifying in spectroscopic data for referencial use.Processor 475 can calculate and measure being positioned at the material that a plurality of probe assembly 410A-410I at buildings 415 diverse location places obtain.If identify a dangerous substance the spectroscopic data obtaining from one or more probe assembly 410A-410I, processor 470 can be reported immediately to alarm and responding system 480.Described dangerous substance for example comprises explosive and fire goods, and poison gas and other harmfuls are learned article, and infective virus and bacterium.Alarm and responding system 480 be for sending early warning notice to wireless router 460, wireless router 460 then by transmission of wireless signals to mobile device 490 and other wireless devices, warning safety and other responsible persons take suitable response activities.Mobile device 490 can comprise portable personal computer, personal digital assistant (PDA), a mobile internet device (MID), a portable phone, a smart mobile phone or a wireless server or router.The example of an application is exactly, on remote monitoring highway through the vehicle in inspection post, thereby the detecting device in inspection post is near any explosive substance in vehicle can not triggering of electromagnetic protection.Inspection behavior can comprise mechanical arm that controlled in wireless is equipped with probe may wired by other (as optical fiber and optical cable) for detection of, analyser with the gas phase sample in collection vehicle or wireless mode be connected on probe.

In certain embodiments; referring to Fig. 4 D; probe assembly network installation is at buildings, airport, customs, goods or baggage conveyor system, healthy advisor's office, check-point, harbour, vehicle, boats and ships, submarine, aircraft, train, subway, buildings, industrial site, holiday resort, shopping mall, research laboratory, school or water source; the pre-position in crowd massing ground etc. place, as above about described in Fig. 3 B-4C.Each probe assembly comprises the probe in a detecting device and detecting device, and probe is for launching a laser beam and collecting the scattered light from testing molecule.Detecting device can have one for adsorbing the nanostructured surface of molecule.Probe assembly also comprises for example, a spectrometer from scattered light generation spectroscopic data (Raman spectrum).Detector network can periodically be captured material (step 510) from the environment of each detecting device.Because detecting device has nanostructured surface, captive material molecule is to be attracted on the nanostructured surface of detecting device.Then from the molecule being adsorbed in the nanostructured surface of one or more detecting devices, obtain spectroscopic data (step 520).Selectable, detected molecule can be by a sample solution capture, and/or is attracted on the nano particle being suspended in sample solution.As mentioned above, by the laser instrument in probe assembly, launch a laser beam irradiation and be adsorbed in the molecule on detecting device nanostructured surface or in sample solution.Light through these molecular scatterings gathers by probe assembly.Spectrometer in probe assembly obtains spectroscopic data from scattered light, for example Raman spectrum.Nanostructured surface on detecting device plays amplification humidification to the signal intensity in Raman spectrum.Capture material and obtain relevant spectroscopic data and can periodically carry out, for example interval is 1 minute, 10 minutes, 15 minutes or several hours.In certain embodiments, can respond and come from the order of control center and produce spectroscopic data.

Spectroscopic data is then from detecting device TCC traffic control center (step 530).The materials capture time will transmit together with spectroscopic data.Spectroscopic data can transmit by wired mode (as shown in Figure 4 A) or cordless communication network (as shown in Figure 4 B and 4C).Control center can comprise that a spectroanalysis instrument and stores the database of the spectral signal of predetermined known danger material.Spectroanalysis instrument is used for measuring whether having spectral signal in the spectroscopic data of self-detector.If found the spectral signal of a known danger material in this spectroscopic data, identify this dangerous substance (step 540).

This dangerous substance may be identified by a plurality of detecting devices in detecting device network.This dangerous substance may be identified by different detecting devices at different time.For example, as the passenger 200-2 walkway 210(Fig. 3 A that passes by), in this network, different detecting devices can obtain the dangerous substance at different time and diverse location place.Processor 475(Fig. 4 B of control center) by the acquisition time of the position of detecting device and spectroscopic data is associated, can measure position and the life period of dangerous substance.By the position of detecting device, can measure the position of a fixing dangerous substance.The position correlation of the dangerous substance that different detecting devices detect can be used as the exact position that weighting factor is measured this dangerous substance, and this can represent with a coordinate system two dimension (2D) or three-dimensional (3D).The different detecting devices of diverse location are used for measuring the space-time distribution situation (that is to say the function of position as the time) of this dangerous substance by the processor of control center to the capture time of dangerous substance.Can predict this dangerous substance position in the future by processor thus.

In certain embodiments, the spectroscopic data of detecting device collection can connect with the graphic materials collecting from spectroscopic detector periphery scene.For example, the video camera 405 that is positioned at the spectroscopic detector side of identifying dangerous substance can be recorded questionable person's thing or goods.The image of suspect or goods be stored and the positional information of the dangerous substance of reporting for work to take suitable responsive measures.

And then to alarm response system, send alarm signal, alarm response system starts this dangerous substance to take responsive measures (step 560).Described alarm signal can be the forms such as Email, text message and voice call.Urgency level can be classified by the different hazard level that for example green (safety), blue, yellow, orange, red (the most dangerous) represent.Alarm signal can comprise the position that dangerous substance is current and/or expect, and with the suspect of dangerous substance or the appearance information of goods.Related personnel receives alarm, sends guardian of the peace to arrive dangerous substance place, starts to evacuate.

Fig. 5 utilizes detector monitors to be discharged into the schematic diagram of the harmful chemical substance in environment.Probe assembly 120 is distributed in the periphery in potentially contaminated source, the near Highway that for example factory 260 or a large amount of automobile 270 pass through.Probe assembly 120 can be distributed in monitored area near, by transmission of stray light, to spectroanalysis instrument 150, spectroanalysis instrument 150 is measured content and the concentration that is discharged into the material in environment.Monitoring sample include but not limited to soil, water, lake, river, beach, well, plant, air, aerosol (gasoloid, aerosol) etc.This application may extend to vehicle exhaust inspection and monitoring, probe assembly is positioned over to the exit of automobile exhaust port.

the application of some spectral detection based on nanostructured

In certain embodiments, the Raman detection system of compact has radio communication performance, can in human body, use.For example, Raman system chip can comprise little lasing light emitter, semiconductor or microminiature spectrometer, wireless module and little probe based on MEMS device etc.An example application is the diagnosis of disease of digestive system.For example, patient can swallow the Raman spectrum detection system of next tablet size after cleaning its digestive system.According to predetermined time interval carry out Raman spectroscopy scans.Spectroscopic data is transferred to the wireless receiver outside human body by a wireless module subsequently.Computer expert cross search and by spectroscopic data be stored in data in database and compare and mate, thereby identify disease.In Another application example, the minimum invasive probe of a needle-like can be carried to the diagnostic region in human body by little Raman detection probe.Raman spectrum data can be transmitted by optical fiber or wireless module.Such application includes but not limited to the diagnosis of cancer (for example breast cancer, colon cancer, cancer of the esophagus, lung cancer, liver cancer, carcinoma of urinary bladder, cancer of pancreas, kidney, oophoroma, carcinoma of mouth, neck and brain cancer, cutaneum carcinoma and cancer of the stomach), Alzheimer's, Parkinson's disease etc.The example that is used for the tumor marker of SERS is hEGF (HER2), CA-125 or CA-549.SERS mark for lung cancer is carcinomebryonic antigen (CEA) or A-549.

Raman spectrum detection system disclosed by the invention and method are suitable for biotechnology and biomedical applications, for example, by detecting A549 cell, DNA, RNA and the protein example of human or animal's tissue or body fluid, lung cancer, and the identity that biomarker (comprise CEA, CA-125, CA19-9, CA-549, PSA, AFP, A549, DNA sequencing, DNA classification, etc.) carries out biostatistics is veritified.

Raman spectrum detection system disclosed by the invention and method are applicable to medicament research and development screening.For the sample of medicament research and development screening, can test and/or breath test obtains by human body fluid.Raman spectrum detection system disclosed by the invention and be equally applicable to criminal investigation.Sample can be liquid phase, as human body fluid or animal body fluid, and the form of saliva, urine, blood, serum or powder for example.Related application also comprises that identification forges a signature, and by DNA type, identifies and examination individual, identifies small paint chipping, fibre identification, etc.

Raman spectrum detection system disclosed by the invention and method are suitable for safety applications, for example, detect dangerous substance, chemical weapons, biopreparate, explosive (powder, solid, liquid, gasoloid or gas form), fire goods (comprising liquid, solid and powder), arcotic and radiomaterial.

Raman detection system and method disclosed by the invention is suitable for food security inspection and environmental monitoring.Can detect the harmful chemicals and the biological that in food, fruit, beverage and water, with gas, liquid, powder, gel, gasoloid or solid form, exist.These harmful chemicals comprise the heavy metal in remains of pesticide (such as acephatemet, cypermethrin, decis, malachite green etc.), bioxin, illegal man-made additive (such as Sudan red 1, Sudan II, s red Ⅲ, Sudan IV, melamine, rhodamine B, sulfide (such as NaS), artificial green etc.), water, include but not limited to compound, prussiate (for example KCN, NaCN), chlorate, sulfate containing Pd, Cd, Hg, As, Cr, Cu metal and these metals.(for example potato block processing temperature is higher than the acrylamide of 120 ℃ of generations can to utilize Raman spectrum detection technique disclosed by the invention to monitor secondary product in food and medicine process, the melamine producing in biochemical drug manufacture process, etc.) for example, to detect harmful chemicals, acrylamide.Food inspection includes but not limited to potato block, chips, fried potato, crisp chrisps, cooky, cracker, bread basket, crisp bread, bread, coffee, refining toast, the nut toasting, biscuit, chocolate, puffed rice and comprises the aquatic products of fish, etc.Medicine investigation includes but not limited to biochemical drug raw material, semi-manufacture or comprises NH ₂the product of base and/or aryl.

Raman spectrum detection system disclosed by the invention and method are applicable to differentiate and detect packaging for foodstuff technique and prepare material, comprise differentiate and screening for microwave food preservative film, film for kitchen use, packaging for foodstuff, food and liquid container, and processing and prepare the polyvinylchloride rope (polyvinylchloride) of material, the material that contains phthalate and polystyrene (PS).

Raman spectrum detection system disclosed by the invention and method are suitable for differentiating ink signatures, the artwork, gasoline of fake and inferior commodities such as medicine, arcotic, Chinese medicine, milk powder, edible oil, wine, tealeaves, cigarette, jewel, banknote, forgery etc.

Raman spectrum detection system disclosed by the invention and method are suitable for commercial production quality management and production safety monitoring.Other applications can comprise the process control of carrying out for the technique of product quality, gas and wet chemistry machining production line and production safety, comprise semiconductor wet chemical process line, air line and space shuttle, ship, boats and ships, submarine production line or the application places relevant with chemistry in refinery, chemical plant, dust free room, etc.

Raman spectrum detection system disclosed by the invention and detector network can be applicable to medical clinic office, Surgical Operating Room, shopping center, holiday resort, buildings, customs, road inspection post, harbour, airport, vehicle, ship, boats and ships, submarine, aircraft, space shuttle, commercial production place, R & D research laboratory, Quality Control Office, institute of education, experiment office and water source, such as surface water, well, underground water etc.

Fig. 6 A is that application surface enhancing Raman scattering utilization detecting device carries out the schematic diagram of matter monitoring for detecting food quality and safety.The place that light scattering probe 100 is placed near food 280, described food can be edible oil, an apple or other fruit, vegetables, or other may be because of transportation, food processing and even the contaminated food of food growth course.In remains of pesticide, veterinary drug, hormone, fertilizer, the illegal additive of food, packaging material for food or shift come or other the molecule of pollutant be adsorbed light scattering probe 100.Objectionable impurities molecule can be adsorbed to nanostructured surface, as the nano particle in colloidal solution or the microchip that comprises Nanosurface structure.As following more details, describe, spectrum detection technique can comprise Surface enhanced raman spectroscopy, normal Raman spectroscopy, and fluorescence spectrum, etc.

Fig. 6 B is that application Raman spectroscopy disclosed by the invention is the schematic diagram that matter monitoring is carried out in early stage disease detection and diagnosis.Doctor can remote monitoring and the patient of diagnosis at home and in hospital.Probe assembly 610 is placed in patient 620 side to carry out physical examination, the inspection of disease recovery situation or medical diagnosis on disease.The air that people breathes out may carry special chemical substance for example alkene and benzene derivative.If the people by examination suffers from disease, cancer for example, include but not limited to lung cancer, breast cancer, liver cancer, cancer of pancreas, oophoroma etc., Raman detection system and method can obtain the finger-print of some chemical substance in breath test, thereby identifies for example cancer of some special diseases.Patient exhales to probe assembly 610.Detecting device in probe assembly receives the air entering, and produces from the corresponding scattered light of molecule in patient or breathe air offering sample person's air-flow.The spectroscopic data of scattered light is produced by spectrometer 630.Wireless communication line 640 is converted into radiofrequency signal by spectroscopic data, and radiofrequency signal is launched radio signal through antenna 645.This radio signal can also comprise patient 620 information (such as patient's name, identity etc.).Terminal 650 and wireless communication line 640 couplings, can show and derive from the information of doctor's office and allow patient's input message to be transferred to doctor's office.By detecting human or animal's body fluid, can carry out similar application.

Antenna 655 in doctor or health consultation office receives the radio signal from a plurality of patients outside certain distance.Wireless server 660 will transform radio signal and take passages patient's spectroscopic data and other relevant informations.Spectroanalysis instrument 670 utilizes the spectral signal being kept in database 680 to analyze spectroscopic data.Spectral signal may show a plurality of predetermined diseases.Spectral signal in spectroscopic data shows that patient suffers from relevant disease or also from the disease of past diagnosis, do not recover completely really surely.Signal intensity can show the order of severity of disease.Doctor 690 can also make the judgement of disease character and the order of severity by inspection spectroscopic data.The system and method that the present invention describes is suitable for early stage medical diagnosis on disease, and described disease includes but not limited to lung cancer, breast cancer, cancer of the stomach, cirrhosis, renal failure, ulcer etc.When testing human body fluid, body fluid manually or is automatically incorporated on detecting device, or Raman detection device is connected on toilet water closet, thus Mining sample at an easy rate, the abnormal signal detecting with Real-Time Monitoring disease and drug effect.This application also comprises to be identified and collating sort protein, DNA and RNA.In above-mentioned application, all test specimens can engage with detecting device to strengthen sensitivity and the intensity that Raman scattering detects.Utilizing Raman scattering to detect micro-trace chemistry material can also be for other field, include but not limited to identify cancer, acquired immune deficiency syndrome (AIDS) (HIV), Alzheimer's, Parkinson's disease, for the non-intrusion type glucose that monitoring diabetes are carried out is tested, for early-stage cancer examination, forbidden drug are monitored the test of non-intrusion type carotenoid levels and the assessment that antioxidant status is carried out.

Fig. 6 C is the schematic diagram that Raman scattering is applied to commercial production quality management, safety assurance or food security in distribution and retail channel.This application can comprise that the monitoring of long-range or unit, the long-range trace chemistry material of chemical concentrate on-line monitoring in wet chemistry machining production line, chemical vessel sealing benchmark check, semiconductor chip defect testing, and food, fruits and vegetables storage monitoring, etc.For example, food can be included in the edible oil of different places sampling.Spectral signal is through the probe collection of a plurality of positions, by optical fiber, by channel transmission, to the spectroanalysis instrument of control center, carries out spectroscopic data analysis.According to the Spectral signal pattern recognition in spectroscopic data, go out the objectionable impurities in the samples such as food, medicine, chemical related substances.

Fig. 6 D is the schematic diagram of a hyperchannel Raman scattering detection system, and it can carry out identification and examination and the food security examination of fake and inferior commodities.This application can comprise the operations such as food, medicine examination, wherein may comprise or not comprise the nanostructured detection module in detecting device.The laser beam of detection system can direct irradiation test specimen.From the scattered light of test material by the collection of popping one's head in.The Raman spectrum of this scattered light demonstrates spectral signal, and this spectral signal can be pointed out whether in these commodity, to have added illegal additive.Potential fake and inferior commodities, for example milk powder, wine and medicine, can be used as to be detected and material examination and be placed under Raman detector.Collected and through hyperchannel, by Optical Fiber Transmission, given the spectroanalysis instrument that is arranged on central office from the spectral signal of different samples, to carry out the analysis of spectroscopic data.This application can extend to signature and the evaluation of banknote, the raman scattering spectrum of signature and banknote generation is compared with the spectrum of legal signature and banknote, thereby check out forgery signature and counterfeit money.

utilize the spectral detection of nano particle

In certain embodiments, referring to Fig. 7 and 8, sample solution 720 is placed in container 710, and container 710 is the cuvette (step 810) of an optical sample bottle or quartz, glass, plastic material for example.Container 710 can be an optical sample bottle, a beaker or a cuvette, etc.Sample solution 720 contains nano particle 750.Nano particle 750 can be present in sample solution 720 with the form of molten colloidal suspension.The reagent that contains chemical substance or biochemical substances is introduced in sample solution 720 (step 820).This reagent can exist with solid, liquid, gasoloid, collosol and gel or gas form.This reagent is dissolved in sample solution 720, allows chemical substance or biochemical substances molecule be adsorbed on the surface of nano particle 750 (step 830).Probe 110(is as shown in Figure 1A) send a for example laser beam of an incident light 701(), irradiate nano particle 750 and chemical substance or biochemical substances (step 840) in sample solution 720.The scattered light 702 coming from nano particle 750 and chemical substance or biochemical substances is gathered (as shown in Figure 1A) (step 850) by probe 110.From the signal of probe assembly output, through spectroanalysis instrument 150, analyze.Detail is shown in example below, from scattered light, obtains a Raman spectrum (step 860).Spectral signal in Raman spectrum can be used for mensuration and is adsorbed to micro-trace chemistry material or the biochemical substances (step 870) on nano particle.

One side disclosed by the invention, the material of the nano particle 750 in sample solution 720 forms wants to strengthen scattered light 702 and from the intensity of the raman spectral signal of nano particle.For example, nano particle 750 comprises metal material (as Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and their alloy), oxide material (as titania, silicon dioxide, zinc paste etc.), silicon and polymeric material.Nano particle 750 can be charged in sample solution 720, and this contributes to separation between nano particle and the formation of colloidal suspension.Nano particle 750 can also comprise the polymkeric substance that is tethered in particle surface, contributes to them in sample solution 720, mutually to repel.

In certain embodiments, nano particle 750 can comprise one by magnetic material as Fe ₂o ₃, Fe ₃o ₄, CoMe, or the core forming containing the compound of Fe, Co or Ni, and the shell coated magnetic core being formed by Au.The diameter of core is within the scope of 1-500nm.After Au shell forms on core, the diameter of core/shell particle is within the scope of about 5nm-50 μ m.Magnetic field in nanoparticle core utilizes outside magnetic field make the separation of particle and gather more effective.Au shell can strengthen the absorption of test substance molecule.And the magnetic field being produced by magnetic core can strengthen resonance and the signal intensity in Raman scattering.

In certain embodiments, nano particle 750 can comprise Ag or SiO ₂core and Au shell.In certain embodiments, nano particle 750 can comprise Ag or Au core and SiO ₂shell.

In certain embodiments, nano particle 750 can comprise carbon nano-tube.The diameter of carbon nano-tube is less than 1,000nm, and for example the diameter of carbon nano-tube can be 0.3-100nm, and length can be that 5nm arrives several millimeters.The length-to-diameter ratio of carbon nano-tube can be up to 5,000 ten thousand.Carbon nano-tube can be walls single wall or many.Carbon nano-tube can be the form of Fu Leti (fullerite), holder shape (torus), nanometer bud (nanobuds) and nanometer flower (nanoflowers).

In system and method disclosed by the invention, carbon nano-tube can be placed in the suspending liquid that sample solution 720 forms a nano particle, and reactant is also added into.Carbon nano-tube can also be introduced in one very on even curface or one have on the surface of nanostructured, and reactant is introduced on this surface of containing carbon nano-tube subsequently.In any situation, laser beam irradiation is on carbon nano-tube and reactant.Strengthen electromagnetic field and can help the electric charge between target chemical substances or biochemical substances molecule to shift, thereby strengthen the signal of Raman spectrum.

Another aspect of the present invention, nano particle 750 can consist of magnetic or ferromagnetic material, for example Fe, Co, Ni, or the compound that contains Fe, Co, Ni, as the alloy of Fe, Co, Ni or oxide, can strengthen raman spectral signal by sample solution 750 is applied to electric field, magnetic field or electromagnetic field like this.This electric field, magnetic field or electromagnetic field can be fix or alternation.

Another aspect of the present invention, sample solution 720 can comprise the potpourri of the nano particle of different composition materials.For example, described nano particle can comprise the potpourri of silicon nanometer or micron particles and metal nanoparticle, or the potpourri of silicon nanometer or micron particles and polymer nano granules, or the potpourri of silicon nanometer or micron particles, metal nanoparticle, metal oxide nanoparticles and polymer nano granules.By the composition of potpourri, can strengthen the intensity of Raman signal.

Another aspect of the present invention, the solvent in sample solution 720 is equally for strengthening the light scattering intensity of nano particle.Ion can significantly strengthen the intensity of Raman signal, and therefore, ionic material is added in sample solution 720.The contained ion of ionic material being added in sample solution 720 can include but not limited to Na ⁺, K ⁺, Li ⁺, Ca ²⁺, Ba ²⁺, Sr ²⁺, Mg ²⁺, Mn ²⁺, Al ³⁺, Zn ²⁺, Sn ²⁺, Sn ⁴⁺, F ^-, Cl ^-, Br ^-and I ^-etc..The ion of sample solution 720 can be unit price, or divalence or more high price.Described ion can be with positive charge or negative charge.Sample solution 720 can have an ionic compound, includes but not limited to LiF, NaF, LiCl, NaCl, KCl, KI etc.Ion concentration can be from 10mM to saturated level.

As shown in Figure 9 A, nano particle 750 can be spherical or irregular shape.Nano particle can be separated from one another in sample solution 720, can be also that gathering is agglomerating.As shown in Figure 9 B, nano particle 750 can have a particle diameter and distribute, and characterizes particle diameter here distribute with average particle size particle size da and particle diameter distribution width dw.Average particle size particle size da can be 1nm-10,000nm, or 2nm-500nm.The value of dw/da can be 0.01-3, and this can limit and singly be distributed to polydisperse distribution of particles.The value of dw/da is generally 0.03-1.

In certain embodiments, sample solution can comprise nano particle and as the micro-section of tumor tissues of reactant.The temperature of sample solution can by a TE refrigeratory and well heater be controlled at one predetermined among a small circle in, temperature variation is less than 1 ℃ or 2 ℃.Temperature can be from-20 ℃ to 60 ℃, or from 0 ℃ to 40 ℃.Dry sample solution on a substrate surface, makes nano particle and reactant stay substrate surface.With this nano particle of laser beam irradiation and reactant.Collection has the reactant of the nano particle of testing molecule to produce scattered light with absorption.By this scattered light, obtain Raman spectrum.Utilize the spectral signal in Raman spectrum can identify chemical substance or the biochemical substances in reactant.

utilize the spectral detection of Nanosurface structure

In certain embodiments, the material that contains micro-trace chemistry material or biochemical substances can be introduced on the surface of chemical detector or detector, and as shown in Figure 1, incident light is scattered, and obtains Raman spectrum to carry out material discriminating.Figure 10-15 have shown the processing step of the nanostructured precious metal surface of a series of making chemical detectors (or detecting device in Fig. 1 105).One sandwich construction 302(Figure 10) comprise a substrate 305, a conductive layer 310 and an alumina layer 315.Substrate 305 can be, for example (the 30-50nm SiO of N-shaped silicon chip (3-8 Ω-cm) or oxidation ₂) p-type silicon (5-10m Ω-cm).Conductive layer 310 can comprise Ti or Ni, and it is deposited in substrate 305, has not only conducted electricity but also heat conduction.The thickness of conductive layer 310 can be optimised, makes it i) adhere to the noble metal film of deposition, for example Ag, Au or Cu film etc. subsequently; Ii) be conductive film, in practical application, sensitive surface applied to electrical bias; Iii) be heat-conducting layer, reduce the temperature of sensitive surface.The thickness of conductive layer 310 generally can be controlled at

scope in, typically,

in scope.

Metal level, for example, aluminium lamination 315 is deposited on conductive layer 310.Aluminium lamination 315 can have 99.999% purity, and thickness is within the scope of 1.0-10.0 μ m.Substrate 305, conductive layer 310 and alumina layer 315 are filling N ₂reacting furnace in 400 ℃-500 ℃ annealing 2-5 hour, make aluminium film recrystallization.Carry out subsequently anodic oxidation, on alumina layer 315, form porous structure, as shown in Figure 11 A and 11B.The porous structure forming on alumina layer 315 comprise many by inwall 314 around hole 312, its cut-open view along horizontal line A-A is as shown in Figure 11 C.In Figure 12, carry out wet oxidation corrosion, to remove the porous Al at top ₂o ₃layer and restraining barrier.Carry out anodic oxidation for the second time and consume all metallic aluminiums, make the Al of the porous at restraining barrier and top ₂o ₃layer is just in time positioned on conductive metal layer.

In Figure 13, carry out restraining barrier reaming that oxide etch is removed hole bottom.Wet etching makes hole 312 extend downward conductive layer.The thickness of formed porous oxide coatings can be controlled by controlling the technological parameter of aluminium physical vapor deposition (PVD), oxidation and wet etching course subsequently.The porous structure of self assembly is formed naturally a hexagonal array.Aperture (d) and interpore distance (D) depend on applied oxidation voltage (V), current density (i) with the character of electrolytic solution, and wet etching reaming process afterwards.

Referring to Figure 14 A, noble metal for example Ag is deposited on porous layer 315 with filling pore 312 and a layer 320 of formation.Layer 320 can form with PVD or plating mode.In Figure 14 B, one deck noble metal 320 is removed, and has retained the noble metal 320-N in hole 312.Again carry out wet method metal erosion or chemically mechanical polishing (CMP) further to control the height of the noble metal 320-N being filled in hole.In Figure 15, the aluminium film 315-AL of aluminium oxide 315 and porous aluminium lamination 315 bottom remnants is removed, and forms a nanostructured surface 300, and it contains a nano-pillar 320-N array.

Nano-pillar 320-N is straight substantially, perpendicular to substrate 305 and conductive layer 310.Nano-pillar 320-N can have substantially the same or close width.Adjacent nano-pillar 320-N is separated by gap, and the distance of these gaps and conductive layer 310 is basic or approach and remain unchanged.

The photoetching of using in above-mentioned manufacturing process matches with the region of the geometric configuration of mask and the dimensional requirement of sensing chip and metal gasket, and metal gasket is positioned on the corner of chip.For field studies, chemicals detects sensing chip and adopts different semiconductor packagings to encapsulate, for example, and Bonding, upside-down method of hull-section construction, system level chip (SOC), etc.

In certain embodiments, nanostructured can be made by different technique, as shown in Figure 16 A-16F.Pair of lamina structure 362 comprises a conductive layer 335 and a substrate 330.Conductive layer 335 can be made by Ti or Ni, can be not only to have conducted electricity but also heat conduction.Substrate 330 can be (the 30-50nm SiO of a N-shaped silicon chip (3-8 Ω-cm) or oxidation ₂) p-type silicon chip (5-10m Ω-cm).The thickness of conductive metal layer 335 can be controlled in scope.Adhesion layer (for example can be made by Ag) can be to be deposited on metal level 335.The thickness of conductive layer 335 can be optimized, thereby micro-trace chemistry material is detected to sensitive surface, applies electrical bias, further, reduces the temperature of sensitive surface for strengthening the sensitivity of micro-trace chemistry material detection, or is clean sensitive surface rising sensitive surface temperature.

In Figure 16 B, layer of precious metal 340 is deposited on the top of conductive layer 335.Noble metal can be a silver layer, the Ag that for example thickness is 10-200nm.In Figure 16 C, the second metal level 345 is deposited on the top of layer of precious metal 340.The second metal level 345 can comprise that purity is about 99.999% aluminium, and thickness is within the scope of 1.0-10.0 μ m.Aluminium lamination 345 is filling N subsequently ₂in reacting furnace, 400 ℃-500 ℃ annealing 2-5 hour, make aluminium film recrystallization.

In Figure 16 D, by oxidation technology, make the porous structure of Woelm Alumina 345 ' form.Figure 16 E is its top view, and this porous structure is formed naturally the hexagon nanoaperture array of self assembly, it comprise many by hexagon hole wall 349 around hole 348.The centre distance of adjacent pores 348 is D.By a wet chemical process, remove after the anodic oxide coating and restraining barrier at top, carry out anode oxidation process for the second time and consume all metallic aluminiums, so that the porous Al at restraining barrier and top ₂o ₃layer 345 ' is positioned on layer of precious metal 340 just.Then carry out wet etching and widen hole 348, and remove the restraining barrier of hole 348 bottoms.While carrying out wet etching, as shown in Figure 16 F, hole 348 is widened, around inwall 349 attenuation of hole.Can control etching process with form a large amount of by inwall 349 around nano-pore 348.Can also corrode between hole 348 and contact each other, produce the hexagonal array of an accurate triangle nano-pillar 349 '.

In Figure 16 G, layer of precious metal 340 is corroded, and hole 348 extends downwardly into the titanium layer 335 of conduction.In Figure 16 H, carry out wet oxidation erosion removal aluminium oxide, succeeded by wet method metal erosion, remove the aluminium that remains in hole 348 bottoms.The aluminium film 315 of aluminium oxide 315 and porous aluminium lamination 315 bottom remnants is removed, and forms a nano column array with controllable height, diameter and intercolumnar distance.This array can have accurate leg-of-mutton periodic hole.

Nano-pillar is straight substantially, perpendicular to substrate 330 and conductive layer 335.Nano-pillar 320-N can have substantially the same or close width.Adjacent nano-pillar is separated by gap, and the distance of these gaps and conductive layer 335 remains unchanged substantially.

In certain embodiments, as mentioned above, being suitable for the preparation of the detecting device of Figure 1A and 1C can be at tool one fixed structure or on without ad hoc structure (being smooth) substrate or introduce nano particle in a sample solution.Micro-trace chemistry material or biochemical substances can first mix mutually with nano particle in solution, the molecule of micro-trace chemistry material or biochemical substances is adsorbed on nano particle, and the sample solution that contains this nano particle is introduced on tool one fixed structure or the surface without ad hoc structure of this chemical detector subsequently.In other words, can produce nanoscale surface structure by the colloidal suspension solution that contains nano particle being coated on to the surface of detecting device 105.Described nano particle can be made by metal material (such as Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and their alloy), oxide material (such as titania, silicon dioxide, zinc paste etc.) or polymeric material.Oxide or polymer beads can or cover with conductive material with metallic ion coating.Colloidal suspension solution can comprise single nano particle or nano particle bunch group.This solution forms nanoscale surface structure after being applied in detector surface.This solution is volatilizable, leaves nano particle target molecule is adsorbed onto to detector surface.

the healthcare appliances of the spectral detection based on nanostructured

In certain embodiments, use as above Figure 1A-2,6B, the relevant described light scattering probe 100 of 7-9B obtains the Raman spectrum of patient's body fluid, can be for the identification of disease by analyzing this Raman spectrum.Human body fluid can be introduced directly into detecting device (as 105 in Figure 1A) above or in containing the sample solution of nano particle (as 720 in Fig. 7) to be mixed.Light scattering and Raman spectrum analysis can be as shown in Figure 1A-1C or Fig. 7 carrying out.In addition, as mentioned above, the sample solution that contains nano particle can transfer to detecting device there is a fixed structure or the surface without ad hoc structure on, subsequently for light scattering and Raman spectrum analysis.

Referring to Figure 17, the Raman spectrum obtaining from a saliva sample from carcinoma of mouth patient has shown the spectrum peak of two features, respectively at 560cm ^-1(at 520cm ^-1-580cm ^-1in region) and 1100cm ^-1(at 1080cm ^-1-1110cm ^-1in region) near, and the healthy individual of not suffering from carcinoma of mouth does not show this two spectrum peaks.560cm ^-1and 1100cm ^-1the characteristic spectrum peak at place and C-S, S-S, O-P-O, PO ₂, the molecular vibration that causes of C-N or C-C key is relevant, for example, contain halfcystine, ATP, ADP, DNA, RNA, protein or other biological samples that contains sulphur or phosphoric acid.The identification of spectral signal can comprise the following steps: in the Raman frequency shift unit with each spectral signal, to be first cm ^-1in the Raman peaks of (wave number), select a spectrum peak; Determine background scattering intensity; Calculate intensity, relative intensity or the integral area at peak.With peak intensity and background, calculate signal to noise ratio (S/N ratio).If signal to noise ratio (S/N ratio) for example, higher than a predetermined threshold (3 or higher), the spectral signal of Raman peaks is just identified.Detection can adopt statistical study and several algorithm (for example Dendrograph and principal component analysis (PCA)) to the identification of the spectral signal of disease and the relevant chemical substance of drug use.If 560cm ^-1and 1100cm ^-1near two spectral signals all identified, can point out so or sentence and think and detect and the Cancer-Related chemical substance in oral cavity, detected person can be prompted or sentence to recognize and suffer from carcinoma of mouth or carcinoma of mouth is early stage.Doctor and detected person should use same or other diagnostic techniques further to detect, and to diagnose, whether suffer from carcinoma of mouth or carcinoma of mouth is early stage.

System and method disclosed by the invention can also be used to test glucose level for assessment diabetic conditions.1115cm ^-1to 1135cm ^-1characteristic spectrum peak in region, for example 1124cm ^-1near, relevant to the molecular vibration of glucose, the Raman spectrum obtaining from the saliva sample of diabetic can provide prompting, sentence and recognize or the key foundation of diagnosing diabetes.The intensity of this Raman peaks, relative intensity or integral area, can be used for the concentration of glucose of evaluating patient body fluid, thereby point out, sentence and recognize diabetes rank.Similarly, referring to Figure 18-20, chemical substance relevant to breast cancer also can be at the approximately 560cm of the Raman spectrum of saliva ^-1and 1100cm ^-1place demonstrates spectral signal (Figure 18).Chemical substance relevant to lung cancer and oophoroma may be at about 745cm of the Raman spectrum of saliva and blood serum sample ^-1place has a spectral signal (about 740cm ^-1-760cm ^-1in scope) (Figure 19 B and Figure 20).745cm ^-1characteristic spectrum peak and the C-S key in protein or phosphate at place, or the O-P-O key in Z-DNA, T-DNA, or the molecular vibration that the atom that contains S, N or P or molecular radical cause is correlated with.Chemical substance relevant to acquired immune deficiency syndrome (AIDS) may be at the 865cm of the Raman spectrum of blood serum sample ^-1-885cm ^-1in region, 870cm for example ^-1near there is a spectral signal (Figure 21).System and method disclosed by the invention can also be used to whether use the detection of forbidden drug, for example heroin, dexoxyn, cocaine, caffeine, morphine, codeine, amphetamine, ephedrine, papaverine, narcotine, codeine (acetyl codeine), methamphetamine hydrochloride (methamphetamine HCl), ketamine hydrochloride (ketamine HCl), codeine phosphate (codeine H ₃pO ₄), dolantin hydrochloride (meperidine HCl claims again pethidine), triazole benzene phenodiazine (triazolam), quinalbarbitone (secobarbital), hypaconitine (hypaconitine), MDMA etc.What Figure 22 showed is the Raman spectrum from a dexoxyn solid (a kind of forbidden drug), the saliva sample of a medication individuality and a dexoxyn user saliva sample.From the Raman spectrum of drug use person's saliva sample at about 1030cm ^-1and 1535cm ^-1near respectively have a characteristic peak, this characteristic peak shows that it may use forbidden drug.Method and system disclosed by the invention can also be used for detecting the excitant (for example hormone) in sportsman's body in international athletic competition (such as the Olympic Games).

Similarly, referring to Figure 23, the state of smoking state and passive smoking also shows spectral signal, at the approximately 1029cm of smoker's saliva sample Raman spectrum ^-1the spectral signal that place demonstrates is not present in the healthy individual of non-smoking.About 1029cm ^-1the characteristic spectrum peak at place is relevant to the molecular vibration pattern of N-first-2-5-pyrrolidone (cotinine), and N-first-2-5-pyrrolidone is the metabolic product of nicotine.

Utilize Raman probe disclosed by the invention to carry out one or more in can comprising the following steps of non-invasive disease detection and diagnosis: referring to Figure 24, first from a patient or forbidden drug user, to obtain body fluid (step 2010).Because Raman scattering detecting device disclosed by the invention has high sensitivity, the amount of body fluid can be quite little.For example, the body fluid volume obtaining from patient can be from about 100pl to 4ml.The example of body fluid can comprise blood, saliva, urine, serum, tear, sweat, gastric juice, hydrothorax, ascites, celiolymph, seminal fluid and juice.Centrifugal rear body fluid is introduced on a Nanosurface (step 2020).For example, this Nanosurface can comprise the nanoscale structures in detector surface.Body fluid can be transferred on the Nanosurface of detecting device.The body fluid of staying detector surface after dry forms a drying layer.In another example, Nanosurface is provided by the nano grain surface being suspended in solution.Body fluid be directed in the solution that comprises nano particle.Molecule in body fluid is adsorbed on Nanosurface.With this Nanosurface of laser beam irradiation be adsorbed to the molecule (step 2030) on Nanosurface.Mining collection is through Nanosurface and the scattered light (step 2040) that is adsorbed molecule.

From scattered light, obtain Raman spectrum (step 2050).One or more spectral signals in spectrum are identified diagnose the illness (step 2060).Can comprise cancer by detected disease example, include but not limited to lung cancer, breast cancer, cancer of the stomach, cancer of the esophagus, thyroid cancer, laryngocarcinoma, ulcerocancer, oophoroma, liver cancer, head and neck cancer, the cancer of the uterus, cervical carcinoma, carcinoma of mouth, leukaemia, colon cancer, carcinoma of urinary bladder, prostate cancer, cutaneum carcinoma, bronchiolar carcinoma, kidney, cirrhosis, renal failure, acquired immune deficiency syndrome (AIDS) and dopy.As mentioned above, one or more spectral signals predetermined Raman frequency shift place in Raman spectrum.Raman frequency shift and spectral signal feature are special for disease to be detected.For example, the spectral signal in carcinoma of mouth and breast cancer saliva sample may be at about 560cm ^-1or 1100cm ^-1near.Spectral signal 745cm in Raman spectrum greatly in Sera of Lung Cancer sample ^-1near.One spectral signal can comprise a spectrum peak.When this spectrum peak is greater than certain predetermined threshold, spectral signal is identified.For example, with respect to noise background, just identified when the signal to noise ratio (S/N ratio) of spectrum peak is greater than 3.

It should be noted that Figure 24 and Fig. 8 are consistent, in the step of Figure 24, can add the one or more steps in Fig. 8, comprise the sample solution utilizing containing nano particle.

spectroscopic methodology sensing detection based on nanostructured in biological immune mensuration

In certain embodiments, disclosed light scattering probe and substance detecting method are applied to biological immune mensuration.Especially, nanostructured strengthens Raman spectrum analysis and utilizes immunoassays, as Enzyme Linked Immunoadsorbent Assay (ELISA) or enzyme immunoassay (EIA) (EIA) are applied to using antibody or antigen as the detection of analyte.

Referring to Figure 25, a kind of material discrimination method can comprise the following steps: known antigens is incorporated in the different antibodies being adsorbed on Nanosurface, to form different Ag-Abs pair in Nanosurface structure.In the Raman spectrum obtaining from each Nanosurface structure, set up Raman signal (step 2510).In certain embodiments, spectrum mark (for example, Raman labels) is as MBA(p-mercaptobenzoic acid) before analyte detects, be introduced into antibody or antigen and be combined to provide the signal in Raman spectrum.For example, the main SERS signal of MBA mark is at 520cm ^-1, 1072cm ^-1and 1588cm ^-1place.Spectral signal is determined by the identical method of the spectral signal in Figure 17-23 with above-mentioned and disease association.A spectral signal can comprise one or more spectrum peaks, one or more spectrum paddy, and other spectral shapes, and as peak relative height, crest line width and peak shape, one or more molecular links that can be used in characterising biological, medical science or chemical substance.Biology, medical science or chemical substance can comprise that can cause human immune system produces antibody.

Antibody, is also known as immunoglobulin (Ig), is the gamma globulin finding in vertebrate blood or other body fluid.They are used for identifying and eliminating foreign matter by immune system, as bacterium and virus.They are representational consists of basic structural unit---and each has two large heavy chains and two little light chains, to form, for example, has the monomer of a unit, has the dipolymer of two unit or has the pentamer of five unit.Antibody is known as plasmacytic Leukocytopoiesis by a class.There are several dissimilar heavy chain of antibody, and several different types of antibody, the heavy chain having based on them is divided into different isotypes.Known in mammal have five kinds of different antibody, and it plays the part of different roles, and the different foreign matter running into for them helps to implement suitable immune response.

Although the general structure of all antibody is closely similar, on albumen top, a little region is to be very easy to change, and allows millions of antibody to have slightly different apex structures or antigen binding site existence.This region is known as hypervariable region.Each of these hypervariable regions can be in conjunction with different targets, namely antigen.The great diversity of antibody allows immune system recognition to equate wide in range multifarious antigen.Unique position that antigen is identified by antibody is known as epitope.These epitopes combine with its antibody with the interaction of high special, are called induced-fit, make antibody identification in conjunction with its unique antigen from form the molecules up to a million of biosome.Identification by antibody to antigen, its attack for other parts of immune system of mark.Antibody also can directly pass through, and for example, causes the required part of infection combine to eliminate object with pathogen.

The antigen that is applicable to method and system disclosed by the invention comprises material, arsenic compound or the prussiate in Clenbuterol, nitrofuran, phosphorus, organochlorine, positive terephthalate, pesticide, rat-bane, veterinary drug.

Target molecule can comprise little molecule and metallic element, as forbidden drug (heroin, dexoxyn, cocaine, caffeine, morphine, codeine, amphetamine, ephedrine, papaverine, narcotine and MDMA, etc.), nitrofuran, Clenbuterol, chloromycetin, Olaquindox and heavy metal objectionable impurities mercury, lead, cadmium.

Should be noted that, although following process prescription unknown antigen as analyte and known antibodies, be to be attached in advance nanostructured surface for detection of antigen, contrary process is also applied in the system and method for the following stated: unknown antibody is as analyte, and known antigens is prepared for being attached to nanostructured surface for the detection of unknown antibody.

In order to detect and identify unknown antigen, a plurality of sample solution 264_1 are provided ... 264_i ... 264_N, as shown in Figure 26 A.Solution 264_1 ... 264_i ... each in 264_N comprises nano particle 265_1 ... 265_i ... the suspending liquid (step 2520) of 265_N.Nano particle 265_1 ... 265_i ... 265_N can be circular or irregularly shaped.Nano particle 265_1 ... 265_i ... the mean diameter of 265_N is in about 1nm to 1000nm scope.Described in Fig. 9 A described above and 9B, nano particle 265_1 ... 265_i ... the Size Distribution of 265_N can characterize with average particle size particle size and particle size distribution width.Nano particle 265_1 ... 265_i ... the material of 265_N can be selected from metal, metal alloy, oxide material, silicon, compound substance and combination thereof.Nano particle 265_1 ... 265_i ... the material of 265_N can be selected from Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and combination thereof.Nano particle 265_1 ... 265_i ... the material of 265_N can be selected from titanium oxide, Si oxide and zinc oxide.Nano particle 265_1 ... 265_i ... 265_N also can comprise carbon nano-tube.In order to strengthen the spectral intensity of Raman scattering, sample solution can comprise multivalent ion.Nano particle 265_1 ... 265_i ... 265_N can comprise magnetic or ferrimagnet.

Referring to Figure 25 and 26B, different unknown antibody 266_1 ... 266_i, 266_N is incorporated into respectively sample solution 264_1 ... 264_i, 264_N, so that different antibody 266_1 ... 266_i, 266_N is adsorbed onto respectively different sample solution 264_1 ... 264_i ... nano particle 265_1 in 264_N ... 265_i ... 265_N upper (step 2530).Different antibody 266_1 ... 266_i ... 266_N can select according to the type of determined antigen.For example, unknown antigen can belong to material, arsenic compound or the prussiate in Clenbuterol, nitrofuran, phosphorus, organochlorine, positive terephthalate, pesticide, rat-bane, veterinary drug.The antibody of these antigens is incorporated into respectively sample solution 264_1 ... 264_i ... in 264_N, each sample solution 264_i receives dissimilar antibody 266_i.

Next, referring to Figure 25 and 26C, comprise namely analyte of unknown antigen 267() reactant be introduced in and contain nano particle 265_1 ... 265_i, the sample solution 264_1 of 265_N ... 264_i, in 264_N, so that unknown antigen 267 combines with the coupling antibody 266_i in one of them solution.Antigen 267 can be a kind of in above-mentioned exemplary substrate.Other examples of antigen 267 can comprise melamine, honey element, sodium cyclohexylsulfamate, sucrose, starch, nitrite, nitrate, the Sudan I, II, III and IV, malachite green, acephatemet, orthene, DDT, DDV, malathion, fenifrothion, decis, cypermethrin, parathion-methyl, phosmet, thimet, Rogor, nitrofuran, furazolidone, 3-amino-2-oxazolidone (AOZ), heavy metal, plumbous, cadmium, mercury, chromium, arsenic, chloromycetin, duomycin, Ciprofloxacin, Enrofloxacin, Clenbuterol, Ractopamine (ractopamine), salbutamol (salbutamol), penicillin (penicillin), bisphenol-A (bisphenol A), phthalic acid two (2-ethyl hexyl) ester (DEHP), 6-benzyl aminopurine (6-benzylaminopurine) or Olaquindox (olaquindox), it is a kind of controlled substance in food.

Reactant can be separated from food.Described food can comprise dairy products, candy, cooky, beverage, wine, meat, seafood, tealeaves, vegetables fresh or canning, fruit, grain, cereal, cornflakes, potato block or the food that contains protein.The example of dairy products comprises milk, milk powder, cheese, cheese cake, yoghourt, ice cream, contains milk candy or cooky.Reactant also can extract from forbidden drug material, as heroin, dexoxyn, cocaine, caffeine, morphine, codeine, amphetamine, ephedrine, papaverine, narcotine and MDMA, etc.

Referring to Figure 25 and 26D, contain unknown antigen 267 and absorption and have different antibodies 266_1 ... 266_i ... the nano particle 265_1 of 266_N ... 265_i ... the sample solution 264_1 of 265_N ... 264_i, each in 264_N, is irradiated (step 2550) by laser beam 2600 respectively.Laser beam 2600 can be launched by the laser aid in probe 110.From sample solution 264_1 ... 264_i ... in 264_N, the scattered light 2601 of each can be gathered by probe 110 respectively.Each sample solution 264_1 ... 264_i ... the Nanosurface of 264_N strengthens Raman spectrum and by spectroanalysis instrument 150, from scattered light 2601, is obtained (step 2560).

From sample solution 264_1 ... 264_i ... in each Raman spectrum obtaining in 264_N, find the right spectral signal (step 2570) of Ag-Ab of combination.Similar to the description of above-mentioned Figure 17, Ag-Ab can comprise step to the identification of spectral signal: selective light bands of a spectrum around from the Raman signal peak of spectral signal first.Determine backscatter strength level.Calculate peak intensity level, relative intensity or complete peak area.By peak intensity and background level, calculate signal to noise ratio (S/N ratio).If signal to noise ratio (S/N ratio) for example, than predetermined critical high (, 3 or higher), the spectral signal of Raman peaks can be by clear and definite identification.

If Ag-Ab is to 268_i(or above-mentioned relevant Raman labels molecule) Raman signal, the Raman spectrum obtaining from one of sample solution 264_i, find, the unknown antigen 267 in reactant can be confirmed as with the nano particle 265_i being adsorbed onto in sample solution 264_i on the antigen that matches of antibody 266_i.Due to the key-lock characteristic between antigen and antibody, the Raman spectrum obtaining from other sample solutions can not demonstrate the right spectral signal of their Ag-Abs separately.In order to increase the interference of accuracy and reduction spectral background noise, spectral signal can carry out verification by obtaining the nanostructured enhancing Raman scattering of sample solution 264_i, and this sample solution has the nano particle that absorption has antibody 266_i and applies together with the reactant that comprises unknown antigen 267.

In certain embodiments, when scattered light is collected, electric field, magnetic field or electromagnetic field for example can be applied to, in the sample solution that comprises Nanosurface structure (nano particle).Electric and magnetic fields can be direct current or alternating current.These have been observed and can have strengthened the signal intensity that Nanosurface strengthens Raman signal.

In addition, can, by finding the Nanosurface of spectral signal to strengthen Raman spectrum, determine the concentration level (step 2590) of the antigen 267 identifying at sample solution (264_i).The concentration of antigen 267 in sample solution, can be as definite in the intensity of spectrum peak, relative intensity or complete peak area by for example spectral signal.

Utilize biological immune mensuration and Raman spectrum analysis can to strengthen by the Nanosurface structure except nano particle the identification of antigen.For example, Nanosurface structure can be included in the nanostructured of constructing or forming in substrate.This nanostructured can be included in projection or the post forming on upper surface of substrate, and the depression forming in upper surface of substrate or hole.Adjacent projection or the mean distance of post can be in 10nm to 1000nm scopes.The mean distance in adjacent recessed or hole can be in 10nm to 1000nm scope.Projection also can be by forming by the introducing of nano particle or deposition at substrate surface.The width range of nano particle is in 1nm to 1000nm scope.

Referring to Figure 27, spectral signal model from be adsorbed onto the structural different Ag-Ab of Nanosurface to the Raman spectrum obtaining (step 2710).Ag-Ab is to by formation that known antigens is combined with the antibody that is adsorbed onto Nanosurface structure.The antigen that is applicable to method and system disclosed by the invention comprises material, arsenic compound or the prussiate in Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane, veterinary drug.

In order to detect and identify unknown antigen, a plurality of nanostructured 285_1 in substrate, are provided ... 285_i ... 285_N, as shown in Figure 28 A (step 2720).Next, as shown in Figure 28 B, different antibody 286_1 ... 286_i, 286_N is incorporated into respectively nanostructured 285_1 ... 285_i, on 285_N, to allow different antibody 286_1 ... 286_i ... 286_N is adsorbed onto nanostructured 285_1 separately ... 285_i ... the surface (step 2730) of 285_N.Absorption can be by introducing antibody 286_1 ... 286_i ... 286_N is before first at nanostructured 285_1 ... 285_i ... 285_N applies moistening solution and is assisted.Different antibodies 286_1 ... 286_i ... 286_N can select according to the type of determined antigen.For example, unknown antigen may belong to material, arsenic compound or the prussiate in Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane, veterinary drug.

Next, referring to Figure 27 and 28B, the reactant that comprises unknown antigen 287 is introduced in nanostructured 285_1 ... 285_i ... 285_N, so that unknown antigen 287 and nanostructured 285_1 ... 285_i ... coupling antibody 266_i on one of 285_N combines.Antigen 287 can be one of above-mentioned exemplary substrate.Reactant can be separated from food.Food can comprise dairy products, candy, cooky, beverage, wine, meat, seafood, tealeaves, vegetables fresh or canning, fruit, grain, cereal, cornflakes, potato block or the food that contains protein.The example of dairy products comprises milk, milk powder, cheese, cheese cake, yoghourt, ice cream, contains milk candy or cooky.Other examples of antigen 287 can comprise melamine, honey element, sodium cyclohexylsulfamate, sucrose, starch, nitrite, nitrate, the Sudan I, II, III and IV, malachite green, acephatemet, orthene, DDT, DDV, malathion, fenifrothion, decis, cypermethrin, parathion-methyl, phosmet, Rogor, nitrofuran, furazolidone, chloromycetin, duomycin, Ciprofloxacin, clenbuterol hydrochloride or Enrofloxacin, Clenbuterol or 6-benzyl aminopurine, can comprise in food by controlled substance.

Reactant can also be separated from forbidden drug material, comprises heroin, dexoxyn, cocaine, caffeine, morphine, codeine, amphetamine, ephedrine, papaverine, narcotine and MDMA.

Be applicable to another of the inventive method exemplary by controlled substance, be KET (Ketamine, ketamine).KET is people and the veterinary medicine that is applied to anesthesia, asthma, antidepression, drug addiction treatment and the treatment of compound pain syndrome.KET is also the main matter in the illicit drugs " KET " (ketalar) of wide-scale distribution, can cause hallucinations and other psychoreactions.KET is a kind of little molecule, can not directly cause the generation of immune response and associated antibodies.But KET can carry out chemical modification, generate p-NH2-KET, make it be attached to protein carrier, as hemocyanin (KLH), bovine serum albumin (BSA), ovalbumin and gamma globulin, etc.These protein carriers of being combined with KET have the performance of antigen and can cause immune response in human or animal body and the generation of antibody.Utilize Ag-Ab to strengthening the detection of Raman scattering to these protein carriers with nanometer, can realize the identification of KET.

Referring to Figure 27 and 28C, comprise unknown antigen 287 and with different antibodies 286_1 ... 286_i ... the nanostructured 285_1 of 286_N absorption ... 285_i ... each in 285_N is irradiated (step 2750) by laser beam 2800 separately.Laser beam 2800 can be launched by the laser aid of popping one's head in 110.From nanostructured 285_1 ... 285_i ... in 285_N, the scattered light 2801 of each can be gathered by probe 110 respectively.Nanosurface strengthens Raman spectrum by each nanostructured 285_1 ... 285_i ... the spectroanalysis instrument 150 of 285_N obtains (step 2760) from scattered light 2801.

Similar to the description of above-mentioned Figure 17 and step 2570 (Figure 25), from nanostructured 285_1 ... 285_i ... in each Raman spectrum obtaining in 285_N, find the right spectral signal of Ag-Ab of combination.If find Ag-Ab to 288_i(or above-mentioned relevant Raman labels molecule the Raman spectrum from nanostructured 285_i obtains) spectral signal, the unknown antigen 287 in reactant can be determined to be the antigen mating with the antibody 286_i being adsorbed onto on nanostructured 285_i.Due to the key-lock characteristic between antigen and antibody, the Raman spectrum obtaining from other sample solutions can not demonstrate the right spectral signal of their Ag-Abs separately.Spectral signal can carry out verification by the nanostructured enhancing Raman scattering obtaining in a plurality of nanostructured 284_i, and this nanostructured 284_i absorption has the antibody 286_i applying together with the reactant that comprises unknown antigen 287.

The concentration level of the antigen 287 identifying in reactant in addition, can utilize and find the Nanosurface of spectral signal to strengthen raman spectroscopy (step 2790).In sample solution, the concentration level of antigen 287 can utilize spectral signal to measure as the intensity of spectrum peak, relative intensity or entire area.

In certain embodiments, the biological immune of the spectral detection of above-mentioned employing based on nanostructured is measured and be can be used for diagnosing the illness.Reactant can be separated from doubtful people's the body fluid of suffering from disease or the gas of breathing.Body fluid can comprise blood, saliva, urine, serum, tear, sweat, gastric juice, seminal fluid or juice.Unknown antibody and relevant disease in reactant are relevant.Relevant disease is selected from cancer, asthma, allergy, cirrhosis, renal failure, leukaemia, HIV, Alzheimer's, Parkinson's disease, diabetes, craving for tobacco, arthritis, cardiovascular disease, SARS and influenza.Cancer can comprise different types, as lung cancer, breast cancer, cancer of the stomach, oophoroma, the cancer of the uterus, cervical carcinoma, carcinoma of mouth, cancer of the esophagus, thyroid cancer, laryngocarcinoma, colorectal cancer, carcinoma of urinary bladder, prostate cancer, bronchiolar carcinoma, cancer of pancreas, leukemia, lymph cancer, osteocarcinoma, the cancer of the brain, neck cancer and cutaneum carcinoma.

When determining that Nanosurface strengthens the signal intensity of Raman scattering, the approaching of nanostructured surface and unknown materials is a very important factor.In certain embodiments, Nanosurface strengthens the Nanosurface structure of using in Raman scattering and carries out pre-service by chemical substance or biochemical substances, to assist test substance to be adsorbed onto in Nanosurface structure.Above-mentioned antibody, as an example of described chemical substance or biochemical substances, can be assisted unknown antigen absorption or be attached to nanostructured surface.Another example of described chemical substance or biochemical substances is antigen, can assist unknown antibody absorption or be attached to nanostructured surface.Usually, suprabasil nanostructured or nano particle can be processed by other chemical substances or biochemical substances, to assist the absorption of unknown materials.These chemical substances or biochemical substances can be adsorbed to Nanosurface, as antibody, or the bonding properties on decorated nanometer surface, the molecular adsorption of Nanosurface unknown materials.These chemical substances or biochemical substances can carry out special selection according to test substance.

An advantage of method and system disclosed by the invention is that unknown antibody can be identified with very high sensitivity.The enhancing of the Raman spectrum analysis being undertaken by Nanosurface structure, can obviously reduce biological immune antigen recognizing is desired in measuring can detectable concentration level.

In certain embodiments, system and method disclosed by the invention is applicable to compound token immunoassays (multiplex labeled-immunoassay).Reporter molecules (Reporter molecules) for example benzenethiol (thiophenol), bipyridyliums derivant (bipyridine derivatives) and cyanopyridine derivative (cyanopyridine derivatives) is adsorbed on different immune colloid gold particle (immune-gold colloid particles).The colloid of mark and antigen are covalently bound in the specific detection substrate that is coated with different antibodies.After washing solvent off, carry out Surface enhanced raman spectroscopy and carry out examining report molecule.The existence of the Raman signal of the reporter molecules of disengaging distinct antibodies substrate, can indicate the right combination of the specific antigen-antibody of substrate surface, thus identification antigen.

In certain embodiments, sandwich structure (a sandwich structure) mainly comprises and is adsorbed onto suprabasil antibody, and the unknown antigen combining with suprabasil antibody.Having the antibody of combination and the colloidal solid of labeled molecule is further bonded on antigen to form sandwich structure by antigen/antibody combination.Labeled molecule has special Raman signal, can from the Surface enhanced raman spectroscopy of sandwich structure, be identified easily.

Sandwich method is generally used for macromolecular detection.Micromolecular compound is known is only combined with an antibody, thereby can not utilize sandwich structure to detect.In the present invention, race detection method (replacing sandwich method) is used to detect micromolecular compound.

Have many micromolecular compounds, as veterinary drug, pesticide, polychlorostyrene phenylbenzene (PCB) and heavy metal etc., on food, environment, in pharmacy with biological sample in be observed.The competitive nanometer enhancing Raman immunologic assay (SERSIA) of micromolecular compound and the party's ratio juris are as shown in figure 29.First, envelope antigen (as analyte carrier protein pairing) is passed absorption or crosslinked action is fixed in SERS substrate to form solid phase antigen.Limited antibody in solid phase antigen in standard solution or sample solution and tested antigenic competition SERS mark (being simultaneously fixed with the nano particle of SERS bioactive molecule and antibody).Clean after substrate, use Ear Mucosa Treated by He Ne Laser Irradiation sample substrate collection of scattered light to generate Raman spectrum.In signal intensity and test, the quantity of antigen is inversely proportional to.

Two researchs have been carried out: the fixing solid phase antigen SERSIA based on nano particle and liquid solid phase antigen SERSIA.

The method relates to the Preparation and characterization of solid phase antigen, comprises selection and the assessment of SERS substrate; The preparation of envelope antigen; Envelope antigen is suprabasil fixing; The sign of solid phase antigen; Select sealing condition to reduce specific adsorption.

The Preparation and characterization of SERS mark comprises the Preparation and characterization of Au colloidal nanoparticles; The formation that SERS active material and Au colloidal nanoparticles are crosslinked; The formation that antibody and Au colloidal nanoparticles are crosslinked; SERS mark characterizes.

Utilize the optimization of the testing conditions of the micromolecular compound that competitive SERSIA carries out to comprise: the selection of solid phase antigen and SERS marker number; Temperature, humidity and the selection of immune response time; The selection of the composition of damping fluid, concentration, ionic strength and pH; The selection of the composition of cleansing solution, concentration, ionic strength and pH.

The condition of SERS signal measuring is selected to comprise: the selection of the portable multiple test condition of surface-enhanced Raman spectroscopy; Collection is measured from the SERS signal of SERS substrate.

The sign of SERSIA method comprises: set up the typical curve of measuring SERSIA sensitivity; Cross reaction tests to measure the specificity of SERSIA; Accuracy and the precision of mark-on sample measuring SERSIA; Stability experiment is to measure the stability of SERSIA.

Actual sample analysis comprises collection, pre-service, target separation, extracts, with SERSIA, measures, to measure yield.

chromatography and spectrum sensing based on nanostructured in biological immune mensuration detect

In certain embodiments, above-mentioned disclosed Light Scattering Probe and material detection side based on nanostructured is applicable to integrated immunochromatography system.

Referring to Figure 30 A-31B, integrated immunochromatography system 3100 comprises that bottom 3110, one chromatoplates 3120, detect film 3130 and an adsorption plate 3140.Envelope antigen 3150 is coated along the p-wire 3155 detecting on film 3130.The example of antigen comprises material, micro-toxin (microtoxin), polychlorinated biphenyl (PCBs), the polynuclear aromatics (PAHs) in Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane, veterinary drug, etc.

Referring to Figure 30 A and 30B, marking nano structure probe 3160 and competitive antigen 3170 are introduced on chromatoplate 3120.As shown in figure 33, marking nano structure probe 3160 is prepared by Raman labels 4-MBA and antibody 3310 being fixed on the surface of nano particle 3300.Marking nano structure probe 3160 is forms of MBA-nano particle-Ab pairing, then uses bovine serum albumin (BSA), and a kind of closed protein that is used for reducing the nonspecific binding site on nano particle is processed.The material that is applicable to nano particle can comprise the gold nanosphere of collaurum, collargol, gold-silver alloy, gold nanorod, depression, etc.

Referring to Figure 30 B-31B, be placed in chromatoplate 3120 marking nano structure probes 3160 and competitive antigen 3170 and diffuse to then to enter and detect film 3130.If there is no competitive antigen in standard solution or sample solution, the antibody carrying on marking nano structure probe 3160 is caught fixing envelope antigen 3150 on the detection film 3130 at the tested survey line of major part 3155 places.On the contrary, for the standard solution that comprises analyte or sample solution, the binding site on special antibody molecule first analyte is occupied, only for envelope antigen 3150, leaves less binding site.Therefore, less marking nano structure probe can remain near detection line 3155 places of envelope antigen 3150.Like this, the concentration of detection line 3155 place's marking nano structure probes 3160 is just inversely proportional to the concentration of competitive antigen 3170.

Figure 32 for example understands the nanostructure probe example used in Figure 30 A-31B and antigen and the envelope antigen of example.

After on the detection film 3130 at marking nano structure probe 3160 and detection line 3155 places, coated envelope antigen 3150 combines, referring to Figure 34, laser 3400 is illuminated be attached to along detection line 3155 be attached to mark on envelope antigen 3150 nanostructure probe 3160.Scattered light 3410 is gathered and is analyzed by said system and method, for the material detection of the Raman scattering based on nanostructured.

Figure 35 for example understands the operation steps utilize the light scattering based on nanostructured that disclosed integrated immunochromatography system carries out and material to detect.Provide a kind of integrated immunochromatography system 3100(as shown in Figure 31 A).Marking nano structure probe 3160 is introduced in (as shown in Figure 31 B) on chromatoplate 3120.Then the chromatoplate 3120 of integrated immunochromatography system 3100 is immersed in containing in the container (such as plastic containers) 3500 of competitive antigenic solution, so that competitive antigen 3170 is attracted in chromatoplate 3120.Being positioned at marking nano structure probe 3160 on chromatoplate 3120 and competitive antigen 3170 diffuses to then to enter and detects film 3130.If the concentration of competitive antigen is zero in plastic containers 3500, most antibody of marking nano structure probe 3160 deliveries are caught the envelope antigen 3155 at tested survey line 3155 places; On the other hand, in plastic containers 3500, the competitive antigen of higher concentration will be captured on the detection film 3130 at detection line 3155 places less marking nano structure probe.Be attached to along the marking nano structure probe 3160 on the envelope antigen 3150 of detection line 3155 and irradiated with laser 3400.The Raman scattering material based on nanostructured that scattered light 3410 adopts said system and method to gather and analyze for the following specifically describes detects.

Figure 36 has shown that the concentration of competitive antigen is at 10-10 ^-6in the time of within the scope of ng/ml, the Raman scattering intensity obtaining from integrated immunochromatography system 3100.Raman scattering intensity is presented at 1074cm ^-1the peak (Raman signal of MBA) at place, this peak is because competitive antigen and detectable antigens are competed antibody combining site limited on marking nano structure probe, and weakens along with the increase of concentration (C in ng/ml).In plastic containers 3500, the concentration of competitive antigen is higher, and the marking nano structure probe 3160 being captured on detection line 3155 is fewer.

Figure 37 has shown the typical curve of target analytes, and for example, Raman scattering intensity is at 1074cm ^-1the peak at place is the function of competitive antigen (for example Clenbuterol) concentration after baseline correction.B ₀it is the Raman scattering signal intensity when competitive antigen concentration is zero.B is the Raman scattering signal intensity in other concentration.Similar to Figure 36, peak intensity weakens as the function of competitive antigen concentration.Figure 38 has shown the photo of different Clenbuterol concentration in the detection film in detection line region.

Figure 39 for example understands the Raman spectrum under different preparation conditions: a) have Immuno gold nano particle (GNP) and the control experiment of MBA mark; B) without the immune GNP of MBA mark; C) without the GNP of the MBA mark of antibody; And d) do not pass through the nitrocellulose filter (NC film) of any processing.Can observe raman scattering intensity at 1074cm ^-1the peak value at place only occurs when MAB marking nano structure probe exists.Do not have MBA mark or antibody in conjunction with time, at 1074cm ^-1the spectrum peak at place is non-existent.Figure 39 clearly proved the SERS signal of observing only come from be fixed on together with antibody on marking nano structure probe and tested survey line 3155 on the Raman labels MBA that catches of envelope antigen 3150.

Figure 40 has shown the repeatability of the Raman scattering intensity peak of integrated immunochromatography system.Each Clenbuterol concentration be 0,0.01 and during 1ng/ml, along detection line 3155(Figure 30 A-31B) the Raman peaks intensity of 10 sampling areas measured.The skew of standard, between 5.0%-5.6%, has shown the high detection repeatability that adopts disclosed integrated immunochromatography system.

Figure 41 has shown the specificity of Raman scattering signal in immunochromatography system integrated under different preparatory conditions.Can observe the A at phenolethanolamine A(Phenylethanolamine), almost there is no cross reaction between Ractopamine and chloromycetin, this is that SERS intensity is almost invariable because they are used as concentration 1.0,10 with during the competitive antigen of 100pg/mL.The competitive inhibition is only that significantly the latter is than the former more remarkable effect in salbutamol and Clenbuterol.

The antigen that is applicable to integrated immunochromatography system and correlation technique comprises multiple micromolecular compound, as the material in Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane and veterinary drug, if corresponding antibody is available.

Other examples of antigen comprise melamine, honey element, sodium cyclohexylsulfamate, sucrose, starch, the Sudan I, II, III and IV, malachite green, acephatemet, orthene, DDT, DDV, malathion, fenifrothion, decis, cypermethrin, parathion-methyl, phosmet, thimet, Rogor, nitrofuran, furazolidone, chloromycetin, duomycin, chloromycetin, Ciprofloxacin, Enrofloxacin, AOZ, heavy metal, plumbous, cadmium, mercury, chromium, Ractopamine, salbutamol, penicillin, bisphenol-A, phthalic acid two (2-ethyl hexyl) ester, 6-benzyl aminopurine or Olaquindox, it can comprise the controlled substance in food.

KET can, by chemical modification to generate p-NH2-KET, allow them to be attached to protein carrier, for example hemocyanin (KLH), bovine serum albumin(BSA) (BSA), ovalbumin and gamma globulin etc.These protein carriers of being combined with KET have antigen performance and can cause the generation of antibody in immune response and human or animal body, are applicable to the detectable antigens in disclosed system and method.

It should be noted that above-mentioned system and method is also applicable to not depart from other changes that spirit of the present invention is done.This system and method is applicable to competitive immunometric assay and non-competitive immunoassay (for example, sandwich structure method).In non-competitive immunoassay, unknown antigen is incorporated into and detects in suprabasil antibody sites.Then the antibody of mark is incorporated into formation " sandwich " structure on antigen.On sandwich structure, carry out Surface enhanced raman spectroscopy to detect the right enhanced spectrum signal of antibody/antigen, thereby identify unknown antigen.

Although the present invention, to be preferred embodiment described at present, is understandable that, this openly can not regard as its restriction.After reading above disclosure, different changes and modification are apparent beyond doubt to those skilled in the art.Therefore, additional claim can be considered to cover all changes and the modification falling in the real spirit and scope of the present invention.For example,, in the discriminating that approaches or be different from " the bad edible oil " and " good edible oil " the Raman frequency shift place of above-mentioned those spectral signals done.

Claims

1. an integrated immunochromatography system, comprising:

One chromatography unit, is provided for receiving marking nano structure probe, and described marking nano structure probe comprises nano particle and is adsorbed onto the antibody on nano particle;

One detects film, comprises envelope antigen, and wherein said chromatography unit is provided for making marking nano structure probe to diffuse through and enter detection film, and the antibody on described nano particle is incorporated on described envelope antigen;

One laser aid, is provided for Emission Lasers with exposure label(l)ing nanostructure probe, and described marking nano structure probe has the antibody being combined on the envelope antigen detecting on film; And

One spectroanalysis instrument, be provided for obtaining the Raman spectrum from thering is the scattered light of the marking nano structure probe that is combined in the antibody on the envelope antigen detecting on film, and identification with antibody-antigen to the spectral signal in the Raman spectrum being associated, thereby realize the detection and Identification of antibody.

2. integrated immunochromatography system as claimed in claim 1, it is characterized in that, described chromatography unit is provided for receiving competitive antigen and makes competitive antigen diffuse through and enter detection film, and described competitive antigen and envelope antigen are competed to be attached on the antibody on nano particle.

3. integrated immunochromatography system as claimed in claim 1, is characterized in that, described marking nano structure probe comprises a spectrum mark being attached on nano particle or antibody.

4. integrated immunochromatography system as claimed in claim 1, is characterized in that, described antigen is selected from Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane and veterinary drug.

5. integrated immunochromatography system as claimed in claim 1, it is characterized in that, described antigen is selected from melamine, honey element, sodium cyclohexylsulfamate, sucrose, starch, the Sudan I, II, III and IV, malachite green, acephatemet, orthene, DDT, DDV, malathion, fenifrothion, decis, cypermethrin, parathion-methyl, phosmet, Rogor, nitrofuran, furazolidone, chloromycetin, aureomycin, Ciprofloxacin, Clenbuterol and Enrofloxacin.

6. integrated immunochromatography system as claimed in claim 1, is characterized in that, the mean diameter of described nano particle is 10-100nm.

7. integrated immunochromatography system as claimed in claim 1, it is characterized in that, the material that described nano particle comprises is selected from metal, metal alloy, oxide, silicon, composite materials, magnetic or ferrimagnet and their combination, or is selected from Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and their combination.

8. integrated immunochromatography system as claimed in claim 1, it is characterized in that, described envelope antigen comprises Clenbuterol, and wherein Clenbuterol is incorporated on OVA molecule, form OVA-Clenbuterol complex, described OVA-Clenbuterol complex is attached to and detects on film.

9. a method of utilizing integrated immunochromatography system identification antibody, comprising:

By a chromatography unit, receive marking nano structure probe, described marking nano structure probe comprises nano particle and is adsorbed onto the antibody on nano particle;

Make marking nano structure probe diffuse through detection film, described detection film comprises envelope antigen;

Antibody on nano particle is attached on envelope antigen;

By Ear Mucosa Treated by He Ne Laser Irradiation marking nano structure probe, described marking nano structure probe has the antibody being attached on the envelope antigen detecting on film;

From the scattered light of the spectroanalysis instrument from marking nano structure probe, obtain Raman spectrum, described marking nano structure probe has the antibody being attached on the envelope antigen detecting on film;

Identification to the spectral signal in the Raman spectrum being associated, realizes the detection and Identification of antibody with antibody-antigen.

10. the method for utilizing integrated immunochromatography system identification antibody as claimed in claim 9, also comprises:

By chromatography unit, receive competitive antigen; And

Make competitive antigen diffuse through detection film, wherein said competitive antigen is provided for being attached on the antibody on nano particle with envelope antigen competition.

11. methods of utilizing integrated immunochromatography system identification antibody as claimed in claim 9, is characterized in that, described marking nano structure probe comprises the spectrum mark being attached on nano particle or antibody.

12. methods of utilizing integrated immunochromatography system identification antibody as claimed in claim 9, is characterized in that, described antigen is selected from Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane and veterinary drug.

13. methods of utilizing integrated immunochromatography system identification antibody as claimed in claim 9, it is characterized in that, described antigen is selected from melamine, honey element, sodium cyclohexylsulfamate, sucrose, starch, the Sudan I, II, III and IV, malachite green, acephatemet, orthene, DDT, DDV, malathion, fenifrothion, decis, cypermethrin, parathion-methyl, phosmet, Rogor, nitrofuran, furazolidone, chloromycetin, aureomycin, Ciprofloxacin, Clenbuterol and Enrofloxacin.

14. methods of utilizing integrated immunochromatography system identification antibody as claimed in claim 9, is characterized in that, the mean diameter of described nano particle is 10-100nm.

15. methods of utilizing integrated immunochromatography system identification antibody as claimed in claim 9, it is characterized in that, the material that described nano particle comprises is selected from metal, metal alloy, oxide, silicon, composite materials, magnetic or ferrimagnet and their combination, or is selected from Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and their combination.

16. methods of utilizing integrated immunochromatography system identification antibody as claimed in claim 9, it is characterized in that, described envelope antigen comprises Clenbuterol, wherein Clenbuterol is incorporated on OVA molecule, form OVA-Clenbuterol complex, described OVA-Clenbuterol complex is attached to and detects on film.

17. 1 kinds of substance identifications, comprising:

Antibody is adsorbed onto in Nanosurface structure;

Introduce unknown antigen to Nanosurface structure so that unknown antigen combines with the structural antibody of Nanosurface;

By the unknown antigen and the antibody that combine in laser beam irradiation Nanosurface structure;

The unknown antigen combining in collection Nanosurface structure and the scattered light of antibody;

In the unknown antigen combining from Nanosurface structure and the scattered light of antibody, obtain Raman spectrum;

In Raman spectrum, find the Raman signal being associated with the unknown antigen combining and antibody;

Unknown antigen based on Spectral signal pattern recognition as the antigen matching with the structural antibody of Nanosurface.

18. substance identifications as claimed in claim 17, is characterized in that, described antigen is selected from material, arsenic compound and the prussiate in Clenbuterol, nitrofuran, organophosphorus, organochlorine, positive terephthalate, pesticide, rat-bane, veterinary drug.

19. substance identifications as claimed in claim 17, it is characterized in that, described antigen is selected from melamine, honey element, sodium cyclohexylsulfamate, sucrose, starch, nitrite, nitrate, the Sudan I, II, III and IV, malachite green, acephatemet, orthene, DDT, DDV, malathion, fenifrothion, decis, cypermethrin, parathion-methyl, phosmet, Rogor, nitrofuran, furazolidone, chloromycetin, aureomycin, Ciprofloxacin, Clenbuterol and Enrofloxacin.

20. substance identifications as claimed in claim 17, also comprise: the identifying and diagnosing human body diseases of the unknown antigen based on disease association connection.

21. substance identifications as claimed in claim 20, is characterized in that, reactant comprises the body fluid from human body.

22. substance identifications as claimed in claim 20, it is characterized in that, described disease is selected from cancer, asthma, allergy, cirrhosis, renal failure, leukaemia, Alzheimer's, Parkinson's disease, diabetes, craving for tobacco disease, arthritis, cardiovascular disease, SARS, influenza and HIV.

23. substance identifications as claimed in claim 17, it is characterized in that, described unknown antigen comprises forbidden drug material, is selected from heroin, dexoxyn, cocaine, caffeine, morphine, codeine, amphetamine, ephedrine, papaverine, narcotine, KET and MDMA.

24. substance identifications as claimed in claim 17, is characterized in that, described unknown antigen comprises protein carrier and the molecule that is attached to the unknown materials on protein carrier, and wherein the identification of protein carrier causes the identification of unknown materials.

25. substance identifications as claimed in claim 24, is characterized in that, described unknown materials comprises KET or Fluorakil 100.

26. substance identifications as claimed in claim 17, is characterized in that, described unknown antigen is extracted from food.

27. substance identifications as claimed in claim 17, while being also included in the scattered light that gathers Nanosurface structure, apply electric field, magnetic field or electromagnetic field to described Nanosurface structure.

28. substance identifications as claimed in claim 17, is characterized in that, described Nanosurface structure comprises that mean diameter is the nano particle of 1-1000nm.

29. substance identifications as claimed in claim 28, it is characterized in that, the material that described nano particle comprises is selected from metal, metal alloy, oxide, silicon, composite materials, magnetic or ferrimagnet and their combination, or is selected from Al, Ag, Au, Cu, Fe, Co, Ni, Cr, Zn, Sn, Pd, Pt and their combination.

30. substance identifications as claimed in claim 17, is characterized in that, described Nanosurface structure forms in substrate.

31. substance identifications as claimed in claim 30, is characterized in that, described Nanosurface structure is included in projection or the column that upper surface of substrate forms, and wherein adjacent projection or column mean diameter are 10-1000nm.

32. substance identifications as claimed in claim 30, is characterized in that, described Nanosurface structure is included in depression or the hole that upper surface of substrate forms.The mean diameter of adjacent recessed or hole is 10-1000nm.

33. 1 kinds of substance identifications, comprising:

Pretreated chemical substance or biochemical substances are applied to Nanosurface structure;

The reactant that introducing comprises unknown materials, to Nanosurface structure, makes the molecule of unknown materials under the assistance of pretreated chemical substance or biochemical substances, be adsorbed onto Nanosurface structure;

Utilize laser beam irradiation absorption to have the Nanosurface structure of the molecule of unknown materials;

Collection is had the scattered light of Nanosurface structure of the molecule of unknown materials by absorption;

From adsorbing the scattered light of the Nanosurface structure of the molecule that has unknown materials, obtain Raman spectrum;

Find the spectral signal being associated with unknown materials in Raman spectrum; And

Utilize the unknown materials in Spectral signal pattern recognition reactant.

34. substance identifications as claimed in claim 33, is characterized in that, described antigen comprises protein carrier and the molecule that is attached to the unknown materials on protein carrier, and described method also comprises:

According to Spectral signal pattern recognition protein carrier, wherein the identification of unknown materials is to carry out according to the identification of protein carrier.

35. substance identifications as claimed in claim 34, is characterized in that, described unknown materials comprises KET or Fluorakil 100.

36. substance identifications as claimed in claim 33, it is characterized in that, described pretreated chemical substance or biochemical substances comprise and are provided for being adsorbed onto the antibody on Nanosurface body structure surface, described unknown materials is the antigen matching with antibody, described spectral signal be adsorbed onto the structural Ag-Ab of Nanosurface to being associated.

37. substance identifications as claimed in claim 33, it is characterized in that, described pretreated chemical substance or biochemical substances comprise and are provided for being adsorbed onto the antigen on Nanosurface body structure surface, described unknown materials is the antibody matching with antigen, described spectral signal be adsorbed onto the structural Ag-Ab of Nanosurface to being associated.

38. substance identifications as claimed in claim 33, is characterized in that, described Nanosurface structure comprises that mean diameter is the nano particle of 1-1000nm or the nanostructured forming in substrate.