CN108267428A - Portable reinforced surface plasma resonance biosensor - Google Patents
Portable reinforced surface plasma resonance biosensor Download PDFInfo
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- CN108267428A CN108267428A CN201611254028.3A CN201611254028A CN108267428A CN 108267428 A CN108267428 A CN 108267428A CN 201611254028 A CN201611254028 A CN 201611254028A CN 108267428 A CN108267428 A CN 108267428A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
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- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
The present invention relates to a kind of portable reinforced surface plasma resonance biosensors, belong to heavy metal analysis field.The present invention provides a kind of GH spr sensors, and the sensor includes waterproof module, wherein, the waterproof module of the sensor is connected, and pass through connectivity port and be connected with external control circuit by adapter with external laser light source.ONP technologies are combined by the present invention with GH spr sensors, and generation is combined or disengaged than heavy metal ion more significant SPR coupling effects in itself using DNA chain and additional AuNPs and sensor surface.Detector is simple in structure easy-to-use, and the DNA probe method used has many advantages, such as that sample volume is small, is not required to mark and isolate and purify, the detection process time is short, and DNA chip is reusable, further reduces costs.
Description
Technical field
The present invention relates to a kind of portable reinforced surface plasma resonance biosensors, belong to analysis and detecting instrument
Field.
Background technology
Seriousness, persistence and the popularity that heavy metal pollution is endangered by it in recent years become the common people concern emphasis it
One, so heavy metal analysis is all particularly significant in medicine, food and environment etc..Based on the portable of GH-SPR-DNA technologies
Heavy metal detector is mainly used for emergent and Site Detection, and target market is mainly the water enterprise for detecting tap water quality, is supervised
Control the environmental protection administration of heavy metal pollution and the supervision department of agricultural environment, agricultural product and food heavy metal pollution.Other are related to
Field further includes the industrial enterprise's (be such as electroplated, be surface-treated) for being related to heavy metal wastewater thereby.
Surface plasma body resonant vibration (Surface Plasmon Resonance, SPR) technology is a kind of important bioid
Learn determination method.However, traditional surface plasmon resonance biosensor is bulky, price costly, portability it is poor and to being
Stability of uniting (such as laser intensity) requires very high so that technique can not be applied to field quick detection.Some laboratories
Surface plasmon resonance biosensor is voluntarily built to reduce cost, but is substantially for laboratory research work, is unsatisfactory for for scene inspection
The condition of survey.In addition, the sensitivity of surface plasmon resonance biosensor is also not up to managed for small-molecule substance of the molecular weight less than 1000
Think level, therefore, trying to explore highly sensitive analysis method has great significance.It is coupled with Goos-Hanchen effects
Applications of surface plasmon resonance (GH-SPR) is a kind of sensor construction method emerging in recent years (referring to Xiaobo, Y.
Deng Goos-Hanchen shift surface plasmon resonance sensor.Applied Physics
Letters,2006.89(26):P.261108-1-3), it is being enhanced by the coupling of two kinds of physical phenomenons and be not easy by
The transducing signal of interference.But existing sensor takes there are still dependence large-scale instrument and equipment, consuming or needs special skill
The problems such as art personnel are operated, it is difficult to adapt to the needs of simple, quick, accurate, sensitive, live tracing detection, and cost compared with
It is high.
Invention content
To solve problems of the prior art, the present invention provides a kind of GH-SPR sensors, and the sensor includes
Waterproof module, wherein, the waterproof module of the sensor is connected, and pass through connectivity port by adapter with external laser light source
It is connected with external control circuit.
Sensor according to the present invention is provided with sensor element in the waterproof module.
Embodiment according to the present invention, the sensor element include but not limited to：Speculum, lens, prism, displacement
It is one or more in amplifier, position sensitive device etc..
According to the present invention, the prism can be the component part of sensing chip.
Preferred embodiment according to the present invention, whole Optical devices of the sensor are closed in waterproof module.
According to the present invention, the waterproof module light transmittance can be preferably lighttight less than 5%, such as less than 1%.
Sensor according to the present invention, wherein the laser light source is adjusted to light beam by micropore.The diameter of the micropore
It can adjust as needed, for example, about 100~about 500 μm, such as from about 150~about 300 μm, can be specifically about 200~about 250
μm。
Embodiment according to the present invention, the light beam are refracted to the plane of incidence of prism, preferably through speculum and/or thoroughly
Mirror is refracted to the plane of incidence of prism.As example, the light beam is refracted to the plane of incidence of prism by speculum and lens.
According to the present invention, the lens can be convex lens.
According to the present invention, the incidence angle of laser beam can be adjusted by adjusting speculum.
The distance in embodiment according to the present invention, the lens to speculum and prism incidence face is separately
1~6 times of mirror focal length, such as 1.2~3.5 times.For example, it can distinguish with the distance in gold-plated prism incidence face from lens to speculum
For 1.5 times of the focal length of lens and 3 times.
The prism may include at least two optical surfaces.Optionally, at least one of described optical surface can be into one
Step includes porous layer.The prism can be the optical prism made of glass.Preferably, the glass can have higher than 1.2,
Such as it is higher than 1.5 refractive index (n).Embodiment according to the present invention, the prism can use the material of higher refractive index, example
Such as the glass of n >=1.6.As example, the SF10 glass of n=1.732 may be used.
At least one optical surface of the prism, as metal film or metal composite film can be coated in detection faces.It is described
Metal film can select the film of one or more compositions in free metal such as gold, silver, platinum, such as be made of metal nanoparticle
Film.The metal composite film can select the composite membrane that free metal is formed with photoelectric activity substance, preferably metal nanoparticle
The composite membrane formed with photoelectric activity substance, to enhance spr signal.
The photoelectric activity substance includes one or more in polyelectrolyte, carbon nanotube and graphene.Form metal
The method of nano-particular film and composite membrane includes spin coating or electrostatic self-assembled.
The thickness of the film can be 10~500nm, such as 30~300nm, can be specifically 40~50nm.It can be with selective membrane
Suitable depth to obtain stronger spr signal.As example, when using water or aqueous solution as working media, the thickness of film can
To be 45~55nm, such as 50nm.When using air as working media, the thickness of film can be 35~45nm, such as 40nm.
According to the present invention, the sensor can be with water, aqueous solution or air etc. for working media.For example, when with aqueous solution
During for working media, the SPR resonant angles of inventive sensor are preferably more than 70 °, such as no more than 60 °, such as can be about 35 °
~about 55 °, for example, about 40 °~about 50 °, for example, about 45 °.
Sensor according to the present invention, laser beam generates GH displacements in prism, and is amplified by multipath reflection.
Further, then amplified displacement can be measured the value of GH displacements again by optical displacement amplifier by position sensitive device.
Preferably, the waterproof module can be prepared by 3D printing method, so as to obtain desired mechanical part configuration.
According to the present invention, can laser light source be emitted by laser light source module.
The laser light source module is single light source double light path module, including the first polarization beam apparatus, the second polarization beam splitting
Device and the first optical chopper (also known as " chopper ") set between the two polarization beam apparatus and the second optical chopper.
According to the present invention, first optical chopper and the second optical chopper form a pair of synchronous reverse phase optical chopper.It is excellent
Selection of land, the synchronous backward optical chopper can be driven by homologous rp pulse.
Preferably, single beam is divided into two light beams of p- and s- polarizations by first polarization beam apparatus.
Preferably, two light beams of p- and s- polarizations pass through the first optical chopper and the second light of above-mentioned synchronous backward respectively
Chopper modulation, so that two light beams alternately reach the second polarization beam apparatus.
The p- and s- polarizes two light beams respectively in the following way by the first optical chopper and the second optical chopper tune
System：When p- light beams by when, s- light beams are blocked；When s- light beams by when, p- light beams are blocked.
Embodiment according to the present invention, it is modulated after two light beams can be with 100~300Hz, such as 150~250Hz, example
Frequency such as 200Hz alternately reaches the second polarization beam apparatus.
Preferably, described two light beams are incorporated into same light path after the second polarization beam apparatus.
According to the present invention, the laser light source module further includes condenser lens, collimator and between condenser lens and standard
Polarization maintaining optical fibre between straight device.
Preferably, the light beam line focus lens after same light path are incorporated into and import polarization maintaining optical fibre, it is then defeated via collimator
Go out laser.
According to the present invention, the polarization maintaining optical fibre uses in the case of without mobile light source module convenient for output, while really
The polarization state for protecting light beam remains unchanged.
According to the present invention, the laser light source module further includes laser diode.The laser diode is used to emit list
Light beam.
According to the preferred embodiment of the invention, the optical component before light beam incidence convex lens is fixed on left optical table
On, light beam is fixed on from the optical component after prism outgoing on right optical table.Preferably, two optical tables respectively can be with
Rotation.For example, rotating left optical table can adjust incidence angle, and rotation angle is 2 times that incidence angle changes；Right optics is rotated to put down
Platform can make the position sensitive device face emergent light of reception outgoing beam.
According to the preferred embodiment of the invention, reach SPR resonance by adjusting the position raising incident beam of convex lens
The accuracy at angle.For example, when convex lens is far from prism, left optical table rotation angle variation is more than incidence angle at prism and becomes
Change, improve the resolution ratio of angular adjustment；When convex lens is close to prism, left optical table rotation angle variation is less than at prism
Firing angle changes, and increases incident angular scan range.
According to the present invention, the external control circuit amplifies for signal and control circuit.
According to the present invention, the sensor further includes the outlet terminal being connect with signal amplification with control circuit.
The signal based on multichannel lock-in amplifier can be used with control circuit in embodiment according to the present invention, signal amplification
Processing circuit.The signal processing circuit can be handled respectively per signal, then by calculating p- and s- light beams each all the way
The respective position at time point, so as to obtain the curve that GH displacements change over time.
Preferably, the signal amplification includes denoising element with control circuit.For example, it is carried out using 16 analog-digital converters
Denoising.
Preferably, the signal processing circuit based on multichannel lock-in amplifier is the signal based on four road lock-in amplifiers
Processing circuit.The circuit per signal independent optimization, denoising all the way and will preferably amplify.
According to the preferred embodiment of the invention, the sensor can be tied with bioprobe, such as oligonucleotide probe (ONP)
It closes, to carry out high-precision detection.For example, the bioprobe can be connected with the film of the prism of the sensor.
Preferably, the bioprobe, as oligonucleotide probe with prism metal film is connected or is passed through by organic group
DNA hybridization effect is connected.The organic group includes but not limited to such as-S- groups, the group containing-S- or other are suitable
Linking group.
Embodiment according to the present invention, the oligonucleotide probe include the single-stranded double-strand for example containing thymidine (T)
It is one or more in oligonucleotides, the double chain oligonucleotide of T-T mispairing or DNAzyme (deoxyribozyme) probe.
As example, the DNAzyme probes can be the Jenner of DNAzyme, oligonucleotides bottom chain probe and DNA modification
Rice corpuscles (AuNPs) is fixed on the probe of film surface by sulfydryl and DNA hybridization effect.It is described solid as further example
It can also be replaced due to the part in the probe of film surface by the alcohols containing sulfydryl, such as 6- sulfydryl hex- 1- alcohol (MCH).
According to the present invention, the resolution ratio of the sensor can be about 10-6RIU。
It is used to detect the use of ion, such as metal ion, particularly heavy metal ion the present invention also provides the sensor
On the way.The ion includes but not limited to such as Hg2+、Ag+、Pb2+、Cu2+In it is one or more.
The present invention further provides a kind of detection methods of above-mentioned ion, include the use of the sensor.
The ion can be metal ion, e.g. heavy metal ion, including but not limited to such as Hg2+、Ag+、Pb2+、
Cu2+In it is one or more.
Detection method according to the present invention, the sensor can be the above-mentioned sensors being combined with bioprobe.
Detection method according to the present invention further includes and contacts the sensor with sample.
According to exemplary embodiment of the subject disclosure, Hg can be utilized2+The height combined with thymidine in DNA (T) is special
The opposite sex, by being rich in single-stranded or T-T mispairing the double chain oligonucleotide of base T in Hg2+T-Hg is formed under mediation pairing2+-T
Duplex structure, and the physicochemical change that DNA change of configuration derives is measured, so as to fulfill to Hg2+Qualitative and quantitative detection.
Alternatively, according to another illustrative embodiment of the invention, each DNAzyme and heavy metal ion can be utilized
There is one-to-one specificity, DNAzyme is activated by heavy metal ion, bottom chain is divided in the catalysis of enzyme chain, detects the widow of generation
Nucleotide change of configuration degree is so as to obtain the concentration of heavy metal ion.
Detection method according to the present invention may also include remaining bioprobe relic, such as oligonucleotides bottom chain probe
Relic is eluted using eluent such as sulfhydryl compound (such as cysteine, glutathione) solution.In this way, fixation can be made
DNAzyme reused.
Advantageous effect
The present invention will realize the integration of GH-SPR sensors, miniaturization, modularization, to improve the stability of system and reliable
Property, while utilize the identification and quantitative analysis of DNA probe technology modification GH-SPR sensor surface realization heavy metal ion, developing
GH-SPR technologies at the scene quickly detect in Practical Approach.It simplifies the system of GH-SPR coupling sensors and is closed
Modularization can just generate reference light paths using common single light source and realize accurately and reliably GH displacement measurements, and passing through more
Signal enhancing is realized in reflection again.The CCD detector used in position sensitive device and other SPR designs all fluctuates very laser intensity
Sensitivity, alternating double light path and four-way the lock-in amplifier design that the present invention uses eliminate the factors such as laser intensity fluctuation to surveying
The interference of amount reduces the requirement to laser light source power, stability and driver and experimental situation, makes low cost and scene
Using being achieved.
ONP technologies are combined by the present invention with GH-SPR sensors, utilize DNA chain and additional AuNPs and sensor
Surface combines or disengages generation than heavy metal ion more significant SPR coupling effects in itself.Detector is simple in structure easy-to-use, makes
DNA probe method has many advantages, such as that sample volume is small, is not required to mark and isolate and purify, the detection process time is short, and DNA
Chip is reusable, further reduces costs.
Description of the drawings
Fig. 1 is the GH-SPR sensor structure schematic diagrames of the embodiment of the present invention 1.
Fig. 2 component arrangement schematic diagrames for light path system in the GH-SPR sensors of the present invention and its optically and mechanically.
Fig. 3 is the external control circuit schematic diagram of the embodiment of the present invention 1.
The curve that Fig. 4 is changed over time for the GH displacements of the embodiment of the present invention 2, the spectral line of wherein BK7+Au (withSPR)
In, larger amplitude is the GH-SPR waveforms without denoising, and the smaller GH-SPR_16D of amplitude is the waveform after denoising；BK7(No
SPR in spectral line), it is TIR (total reflection) waveform without denoising that amplitude is larger, the smaller TIR_ for after denoising of amplitude
16D waveforms.
Fig. 5 is the detection method schematic diagram based on ONP in the embodiment of the present invention 3 and 4.Wherein, (A) is passes through T-Hg-T
Detect mercury ion；(B) it is that lead ion etc. is detected by DNAzyme endonuclease reactions.
Fig. 6 is influence of the golden film thickness to spr signal in (A) air and (B) aqueous medium in the embodiment of the present invention 5.
Fig. 7 is the response diagram that GH displacements change NaCl concentration in the embodiment of the present invention 6.
Specific embodiment
With reference to specific embodiment, the present invention is further explained.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.Furthermore, it is to be understood that after recorded content of the invention has been read, this field skill
Art personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within limited range of the present invention.
Unless otherwise indicated, the reagent in following embodiments and instrument for commercially available product or can pass through art technology
Method is voluntarily prepared known to personnel.
The preparation of 1 sensor of embodiment
As shown in Figure 1, all optical components used in sensor are all just placed and are closed in a lighttight waterproof
In module, it is connected respectively with the laser light source of outside and circuit board by an adapter and connectivity port.Two two poles of laser
The single beam size that pipe is sent out respectively is limited to about 200 microns by a micropore, respectively by p- and s- polarizers, and inverted light
The modulation of chopper, i.e. p- light beams by when, s- light beams are blocked, and vice versa, and two light beams are with about 240Hz
Frequency alternating, by one by speculum, polarization splitting prism and a convex lens group into optical frames combine, two light beams
Same light path is merged into, then line focus lens are refracted to the plane of incidence for the gold-plated prism for forming sensor chip.From lens to
The distance in speculum and gold-plated prism incidence face is respectively 1.5 times and 3 times of the focal length of lens.Prism material is using high refractive index
SF10 glass (n=1.732), it is 45 ° or so to make the SPR resonant angles that aqueous solution is medium, convenient for the compact placement of optical device
And adjusting.Light beam generates GH displacements in prism, and the size of GH displacements is measured by position sensitive device.
As shown in Fig. 2, the optical component before light beam incidence convex lens is fixed on left optical table, light beam goes out from prism
Optical component after penetrating is fixed on right optical table, and two optical tables can respectively rotate, and rotating left optical table can
Incidence angle is adjusted, and rotation angle is 2 times that incidence angle changes, conducive to the fine tuning of incidence angle.Rotating right optical table can make to connect
Receive the position sensitive device face emergent light of outgoing beam.
As shown in figure 3, in the data acquisition and procession circuit of GH-SPR sensors, since position sensitive device is to pass through both ends
Photoelectric current calculate the position of light beam, and this GH-SPR works alternatively pattern using dual-beam, so one shares four road signals
It needs to handle.It, will be per signal independent optimization denoising all the way using the unique signal processing circuit based on four road lock-in amplifiers
Amplification, then calculate the respective position of p- and s- light beams at every point of time in real time by computer, it is shown in Fig. 4 so as to obtain
The curve that GH displacements change over time.
2 individual reflection GH-SPR of embodiment is tested
Using the sensor of embodiment 1, SPR condition (golden film thickness 50nm, optical maser wavelength 650nm, the incidence of optimization are utilized
45 degree of angle), individual reflection GH-SPR tests have been carried out, have measured significant GH displacements.As shown in figure 4, after SPR is coupled, this
The GH displacements of the sensor of invention significantly increase, and significantly reduce noise using 16 analog-digital converters.Also, system installs additional
After black shell, environmental disturbances are shielded, signal quality significantly improves.
Embodiment 3 utilizes the double-stranded DNA detection Hg containing T-T mispairing2+Detection method
As shown in Fig. 5 (A), a single stranded DNA is fixed on GH-SPR sensor gold film surfaces by sulfydryl, with T-T mispairing
Another single stranded DNA can not form the double-stranded DNA for stablizing hybridization, but have Hg2+In the presence of, with Hg2+Form multiple T-Hg-T
Coordination structure forms the SPR response different from a single stranded DNA in sensor surface so as to obtain stablizing the double-stranded DNA of hybridization,
So as to bring the variation of GH displacements.The length of DNA chain, the number of T-T mispairing are the key that such DNA is designed with position, to this
The optimization of a little parameters will improve detection result.The DNA and Hg being captured2+It can be eluted with cysteine solution, so as to fulfill fixation
It is incorporated in the reuse of the ONP of sensor surface.After tested, detection limit can meet drinking water standard down to 2nM.
Probe and detection method of the embodiment 4 based on DNAzyme endonuclease reactions
As shown in Fig. 5 (B), in step [a]-[c], the AuNPs of DNAzyme, oligonucleotides bottom chain probe and DNA modification
It passes sequentially through sulfydryl and GH-SPR sensor gold film surfaces is fixed in DNA hybridization effect, with 6- sulfydryls hex- 1- alcohol (MCH) part
Replace surface-probe, to reduce hybridization steric hindrance.When adding in metal ion to be measured, oligonucleotides bottom chain probe is cut
It is disconnected, coupled AuNPs is caused to leave the signal so as to generate GH change in displacement simultaneously.Remaining oligonucleotides bottom chain probe
Relic can be eluted in step [e] by glutathione solution, and fixed DNAzyme is enable to reuse.After tested, lead ion
Detection limit meets drinking water standard less than 10nM.
Influence of the 5 golden film thickness of embodiment to (A) air He (B) aqueous medium spr signal.
To optimize SPR condition, respectively using air and water as working media, using Fresnel models to medium/golden film/rib
The SPR effects at mirror interface are simulated, and have obtained the relationship of the optimization of SPR resonant angles and signal strength with golden film thickness, knot
Fruit is as shown in Figure 6.It can be seen that SPR resonant angles are influenced by working media and golden film thickness etc., the position of the invention by adjusting convex lens
It puts and improves the accuracy that incident beam reaches SPR resonant angles.When convex lens is far from prism, left optical table rotation angle variation
Change more than incidence angle at prism, improve the resolution ratio of angular adjustment；When convex lens is close to prism, left optical table rotation angle
Degree variation is less than incidence angle variation at prism, increases incident angular scan range.
6 refractometry of embodiment is tested
Using the sensor of embodiment 1, the performance that refractive index is measured it in NaCl solution is tested.As a result see
Fig. 7.According to NaCl concentration and the relationship 8.73*10 of refractive index-3RIU/M, the resolution ratio of GH-SPR sensors of the invention for~
10-6RIU, better than existing Table top type SPR instruments.
More than, embodiments of the present invention are illustrated.But the present invention is not limited to the above embodiments.It is all
Within the spirit and principles in the present invention, any modification, equivalent substitution, improvement and etc. done should be included in the guarantor of the present invention
Within the scope of shield.
Claims (10)
1. a kind of GH-SPR sensors, the sensor includes waterproof module, wherein, the waterproof module of the sensor, which passes through, fits
Orchestration is connected, and pass through connectivity port and be connected with external control circuit with external laser light source；
Preferably, it is provided with sensor element in the waterproof module；The sensor element includes but not limited to：Speculum,
It is one or more in lens (such as convex lens), prism, displacement amplifier, position sensitive device.
2. sensor according to claim 1, wherein whole Optical devices of the sensor are closed in waterproof module
It is interior；
The waterproof module light transmittance can be preferably lighttight less than 5%, such as less than 1%.
3. sensor according to claim 1 or 2, wherein the laser light source is adjusted to light beam by micropore.It is described micro-
The diameter in hole can be adjusted as needed, for example, about 100~about 500 μm, such as from about 150~about 300 μm, can be specifically about
200~about 250 μm；
Preferably, the light beam is refracted to the plane of incidence of prism, and the incidence of prism is refracted to preferably through speculum and/or lens
Face；For example, the light beam is refracted to the plane of incidence of prism by speculum and lens.
4. according to claim 1-3 any one of them sensors, wherein the lens to speculum and prism incidence face away from
From 1~6 times separately for the focal length of lens, such as 1.2~3.5 times；For example, from lens to speculum and gold-plated prism incidence
The distance in face can be respectively 1.5 times and 3 times of the focal length of lens；
The prism may include at least two optical surfaces；Optionally, at least one of described optical surface can be wrapped further
Include porous layer；
Preferably, the optical component before light beam incidence convex lens is fixed on left optical table, and light beam is after prism outgoing
Optical component be fixed on right optical table；Preferably, two optical tables can respectively rotate；
For example, rotating left optical table can adjust incidence angle, and rotation angle is 2 times that incidence angle changes；Right optics is rotated to put down
Platform can make the position sensitive device face emergent light of reception outgoing beam；
For example, when convex lens is far from prism, left optical table rotation angle variation is more than incidence angle variation at prism, improves angle
Spend the resolution ratio adjusted；When convex lens is close to prism, left optical table rotation angle variation is less than incidence angle variation at prism,
The incident angular scan range of increase；
The prism can be the optical prism made of glass；Preferably, the glass can have higher than 1.2, such as be higher than 1.5
Refractive index (n)；For example, the prism can use the glass of n >=1.6, such as the SF10 glass of n=1.732.
5. according to claim 1-4 any one of them sensors, wherein at least one optical surface of the prism, is such as detected
Metal film or metal composite film can be coated on face；
The metal film can select the film of one or more compositions in free metal such as gold, silver, platinum, such as by metal nano
The film that particle is formed；
The metal composite film can select the composite membrane that free metal is formed with photoelectric activity substance, preferably by metal nanoparticle
The composite membrane formed with photoelectric activity substance；
The photoelectric activity substance includes one or more in polyelectrolyte, carbon nanotube and graphene.
6. sensor according to claim 5, wherein the thickness of the film can be 10~500nm, such as 30~300nm,
Can be specifically 40~50nm；For example, when using water or aqueous solution as working media, the thickness of film can be 45~55nm, such as
50nm；When using air as working media, the thickness of film can be 35~45nm, such as 40nm；
The SPR resonant angles of the sensor are preferably more than 70 °, such as no more than 60 °, such as can be about 35 °~about 55 °, example
Such as from about 40 °~about 50 °, for example, about 45 °.
7. according to claim 1-6 any one of them sensors, wherein emitting laser light source by laser light source module；
The external control circuit amplifies for signal and control circuit；Preferably, the signal amplification includes going with control circuit
It makes an uproar element, such as 16 analog-digital converters is used to carry out denoisings.
8. according to claim 1-7 any one of them sensors, wherein the sensor and bioprobe, as oligonucleotides is visited
Needle (ONP) combines, such as the bioprobe can be connected with the film of the prism of the sensor；
Preferably, the bioprobe, as oligonucleotide probe be connected with prism metal film by organic group or by DNA it is miscellaneous
Friendship effect is connected；The organic group includes but not limited to such as-S- groups, the group containing-S- or other suitable connections
Group；
The oligonucleotide probe includes single-stranded double chain oligonucleotide, the double-strand widow's core of T-T mispairing for example containing thymidine (T)
It is one or more in thuja acid or DNAzyme (deoxyribozyme) probe；The DNAzyme probes can be DNAzyme, few nucleosides
The gold nanoparticle (AuNPs) of sour bottom chain probe and DNA modification is fixed on the spy of film surface by sulfydryl and DNA hybridization effect
Needle；
Preferably, the part being fixed in the probe of film surface can also be by the alcohols containing sulfydryl, such as 6- sulfydryl hex- 1-
Alcohol (MCH) replaces.
9. claim 1-8 any one of them sensor is used to detect ion, such as metal ion, particularly heavy metal ion
Purposes；
The ion includes but not limited to such as Hg2+、Ag+、Pb2+、Cu2+In it is one or more.
10. a kind of detection method of ion includes the use of claim 1-8 any one of them sensors；
The ion can be metal ion, e.g. heavy metal ion；Preferably, the ion includes but not limited to such as Hg2 +、Ag+、Pb2+、Cu2+In it is one or more；
Preferably, the method, which is further included, contacts the sensor with sample；
Preferably, the method may also include remaining bioprobe relic, as the chain probe relic use of oligonucleotides bottom is washed
De- liquid such as sulfhydryl compound (such as cysteine, glutathione) solution elution.
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