WO2005033335A2 - Micro-capteurs et nano-capteurs d’especes chimiques et biologiques a plasmons de surface - Google Patents
Micro-capteurs et nano-capteurs d’especes chimiques et biologiques a plasmons de surface Download PDFInfo
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- WO2005033335A2 WO2005033335A2 PCT/FR2004/050494 FR2004050494W WO2005033335A2 WO 2005033335 A2 WO2005033335 A2 WO 2005033335A2 FR 2004050494 W FR2004050494 W FR 2004050494W WO 2005033335 A2 WO2005033335 A2 WO 2005033335A2
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
- G01N21/553—Attenuated total reflection and using surface plasmons
- G01N21/554—Attenuated total reflection and using surface plasmons detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/648—Specially adapted constructive features of fluorimeters using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N2021/258—Surface plasmon spectroscopy, e.g. micro- or nanoparticles in suspension
Definitions
- the present invention relates to new sensors for chemical and biological species with localized surface plasmons.
- Surface plasmons are electromagnetic modes propagating on a metallic conductive surface and which are the consequence of a longitudinal oscillation of the charge density due to the collective movement of the electrons located on the surface of the metal.
- Surface plasmons can be optically excited. Resonances are thus highlighted which are directly linked to the various modes of propagation of surface plasmons.
- Surface plasmons which can be excited are said to be "delocalized” or “localized”. They are distinguished mainly by their propagation length. The most frequent uses, in the current state of the art, preferentially relate to delocalized surface plasmons which propagate over distances greater than the wavelength of the exciting light, typically of the order of 0.5 micrometer in the visible range.
- Localized surface plasmons are waves which propagate over distances less than the wavelength of the excitatory light, that is to say over submicron or even nanometric distances.
- Surface plasmons are highly sensitive to various characteristics of the surrounding environment. It is thus possible to determine, for example, the physicochemical properties of one or more molecules or nano-objects deposited on a metal surface on which propagates a previously excited surface plasmon and thus to produce chemical and biological detectors .
- a reading beam is transmitted through one of the oblique faces of a glass prism with total internal reflection. This beam is partially reflected on the glass / metal interface towards a detector, part of the energy of the beam being absorbed by the metal.
- the total reflection taking place at the metal / air interface is accompanied by the formation of an evanescent wave which excites a surface plasmon at this same interface.
- the excitation of the surface plasmon can only be done under certain angles of illumination.
- the intensity of the light beam collected by the detector decreases due to the energy transferred to the plasmon.
- a deep minimum is formed in the intensity of the light beam collected by the detector.
- This angle very strongly depends on the profile of the refractive index of the metallized surface, in the thickness of the evanescent field. This refractive index changes depending on the substances adsorbed on the metal layer.
- the resonance angle corresponding to the formation of a surface plasmon, is therefore representative of the adsorbed substances. It is also possible, at a fixed angle of incidence, to adapt the excitation wavelength to the plasmon resonance.
- the present invention has been made to improve the resolution of existing chemical or biological species sensors. It uses studs distributed on the surface of a support and capable of immobilizing chemical or biological species. The size and shape of the pads, as well as their distribution, can be provided to allow resolution at the nanometer scale.
- the localized surface plasmons are particularly used. From a general point of view, according to the invention, the changes in the characteristics of the surface plasmons due to a change in the optical properties of the medium are highlighted. surrounding as a result of adsorption of chemical or biological species on metallic substrates.
- the biochemical species adsorbed on the pads are identified by enhanced surface Raman spectroscopy, this enhancement and therefore this type of spectroscopy being possible thanks to the plasmon resonances of the metal pads.
- the subject of the invention is therefore a micro-sensor or a nano-sensor of chemical or biological species with surface plasmon, characterized in that it comprises studs distributed on the surface of a support, the studs comprising at least an electrically conductive material and being capable of immobilizing said chemical or biological species, the pads having a dimension of less than 1 ⁇ m.
- the nano-sensors are defined as those whose studs have a dimension less than 0.5 ⁇ m (dimension corresponding approximately to the experimental diffraction limit of an optical system) and the micro-sensors are those whose studs have a dimension greater than 0.5 ⁇ m.
- the pads are distributed on the surface of the support according to a two-dimensional matrix.
- the studs can have a cross section (that is to say in a plane parallel to the surface of the support) in the form of a circle or ellipse. If the sensor is a micro-sensor, the section of the pads has its largest dimension between 0.5 ⁇ m and 1 ⁇ m.
- the section of the studs has its largest dimension less than 0.5 ⁇ m.
- the micro-sensor or the nano-sensor can comprise at least two arrays of pads, the shape of the section of the pads of one of the networks being different from the shape of the section of the pads of the other network.
- the electrically conductive material of the pads can be gold or silver.
- the studs can be formed by the superposition of at least two different metallic layers. They can also be formed by the superposition of a metallic layer integral with the support and an ultrathin layer (a few nm thick) of a material allowing the attachment of chemical or biological species.
- the surface of the support can be a surface of a material chosen from dielectric materials, semiconductor materials and metallic materials.
- the micro-sensor or the nano-sensor further comprises means making it possible to increase the sensitivity of the sensor.
- These means may comprise a thin metallic film directly deposited on said surface of the support.
- a thin dielectric layer can be interposed between the metallic thin film and the pads in order to adjust the plasmon resonance according to the thickness of the dielectric layer.
- These means may include a planar waveguide intended to convey a guided electromagnetic mode, this planar waveguide being made on the surface or under the surface of the support and under the studs. They can be constituted by the grouping of studs, the distance separating these grouped studs being sufficiently small to allow electromagnetic coupling between the grouped studs.
- the means making it possible to increase the sensitivity of the sensor may comprise at least one particle associated with a pad.
- This particle can be a particle attached to said chemical or biological species. It can be attached to an object intended to be placed near a stud. This object may be the tip of a near field optical microscope.
- This particle can be metallic, the sensitivity is then reinforced by the coupling between the plasmon resonances of the plot and of the particle. It can be made of a fluorescent material, the fluorescence emission then being exacerbated by the plasmon resonance of the corresponding pad.
- - Figure 1 is a view illustrating the operating principle of a micro-sensor or a nano-sensor according to the invention
- - Figure 2 is a view in perspective of a micro-sensor or a nano-sensor according to the present invention
- - Figure 3 is a view grouping together other alternative embodiments of a nano-sensor according to the present invention
- - Figure 4 is a top view of a micro-sensor or of a nano-sensor according to the present invention
- - Figure 5 is a top view of another micro-sensor or of another nano-sensor according to the present invention .
- FIG. 1 illustrates the operating principle of the invention while FIG. 2 is a perspective view of a sensor according to the present invention.
- Metal studs 2 for example gold or silver, are formed on the surface of a support 1.
- the support 1 can be of any kind: dielectric material (for example glass), semiconductor (for example silicon) or metallic (for example a thin layer of gold deposited on a glass slide).
- the studs are distributed according to a matrix dimensional. They are capable of adsorbing, on their upper face, chemical or biological species such as strands of DNA.
- the studs 2 can be cylindrical studs of 0.5 to 1 micrometer in diameter, separated center to center by distances of the order of a few ⁇ m to a few hundred ⁇ m (for example from 5 ⁇ m to 300 ⁇ m).
- the thickness of the pads can be between 20 and 500 nm.
- the diameter of the studs is generally less than 0.5 ⁇ m and their center to center distance can be between 0.5 ⁇ m and 0.5 ⁇ m.
- the thickness of the pads can be between 10 nm and 100 nm.
- the lighting 5 of the surfaces of the metal studs to be studied as well as the detection of the optical signals coming from these studs are carried out either by a confocal optical microscope, however on a non-exclusive basis, preferably in the case of studs of micron size, or by a near field optical microscope or local probe (SNOM for "Scanning Near Field Optical Microscope”).
- Particular lighting parameters polarization, angle of incidence, wavelength of the excitation light source
- a Titanium-Sapphire laser emitting pulses of 150 fs at a wavelength of 800 nm, can be coupled to a photonic crystal fiber with a core diameter of 3 ⁇ m and create a continuum of white light with 200 mW of power.
- a wavelength analysis of each plot allows a plasmon signature of the plot concerned with or without adsorbed species.
- a reference spectrum is carried out above a blank plot of any adsorbed species.
- a second spectrum is carried out after adsorption of the species. The spectral shift between the two plasmon resonances makes it possible to detect the presence and diversity of chemical or biological species adsorbed on each plot, as well as to assess their concentration.
- the study of the complete sample can be done either by scanning the light beam above the fixed sample, or by scanning the sample under the fixed light beam.
- Exalted Raman spectra obtained by a Raman spectroscopic analysis performed above each plot allow the identification of the chemical species adsorbed on the pads.
- Metallic particles 4 such as, for example, without limitation, gold or silver spheres of a few nanometers in diameter, integral with some of the biological or chemical species to be tested can be used as markers. These particles 4 increase the sensitivity of the detection by reinforcing the wavelength shift of the plasmon resonances by coupling the localized plasmons of these particles with those of the corresponding pads and by improving the signal to noise ratio of the detection.
- a network of studs according to the invention is lithographed on a substrate of approximately 1 ⁇ 1 mm 2 of surface comprising 10,000 cylindrical studs of sub-micromic diameter, 200 nm in height, spaced center to center. of 10 micrometers.
- Figure 3 is a view of other alternative embodiments of a nano-sensor according to the present invention. It is a localized surface plasmon sensor structure particularly suitable for the sub-micron characterization of chemical or biological objects.
- the nanosensor shown diagrammatically in FIG. 3 consists of a network of very small metal nanopots 12 formed on a substrate 11 and on which the species 13 to be detected are adsorbed.
- the network of studs according to the invention is lithographed on a substrate of approximately 10 x 10 ⁇ m 2 of surface comprising 400 cylindrical pads 50 nm in diameter, 20 nm in height and spaced center to center by 500 nm.
- These pads are preferably produced by the electronic lithography technique (making PMMA pads by exposure to electrons followed by metallization, and finally by a "lift-off").
- FIG. 5 show respectively a network of cylindrical studs (cylinders of revolution) in gold 22 of diameter 100 nm and height 70 nm, spaced center to center by 300 nm, and a network of studs with elliptical section, in gold 32 50 nm high, 65 nm major axis and 40 nm minor axis, spaced between 150 nm minor axes and spaced between 200 nm major axes. It is possible to adjust the wavelengths of the plasmon resonances by modifying the size and / or the shape of the pads. Adjusting this resonance wavelength to the excitation wavelength of a laser makes it possible to increase the detection sensitivity in the case of identification of the biochemical species by surface-enhanced Raman spectroscopy.
- the sensor can also be made up of several networks of nanoplot particles produced on the same substrate, each network having its own geometrical characteristics. For example, without implied limitation, the networks of FIGS. 4 and 5 can be produced on the same substrate. Thus each network will have its own plasmon signature at a defined wavelength.
- the resonance wavelength of each grating can be adjusted to the wavelength of several lasers to identify the species by Raman spectroscopy or to the wavelength of absorption or fluorescence emission of several markers.
- Nanoplots cylindrical in shape (with a circular or elliptical cross section), without limitation, may have a multilayer structure in order to allow the grafting of molecules which could not be directly grafted onto a metal surface or in order to '' increase the sensitivity and / or tunability in wavelength of the sensor.
- (grafting of molecules) for example, without limitation, a cylindrical stud of
- 100 nm in diameter can consist of two layers, a lower layer of 50 nm of gold and an upper layer of 3 nm of silicon.
- a cylindrical pad 100 nm in diameter may consist of two layers metallic, a lower layer of 20 nm of silver and an upper layer of 10 nm of gold.
- Metal particles 14, for example without limitation gold spheres whose diameter is typically a few nm, can be attached to the chemical or biological species themselves to increase the sensitivity of detection by coupling between the plasmon resonances studs 12 and those of the metallic particles. Specific supports on which the pads are deposited can also increase the sensitivity of the sensor by coupling between the pads and a guided electromagnetic mode.
- metal studs can be deposited on the surface of a planar or confined waveguide 17 or on a thin metal film having resonances associated with the excitation of surface plasmons.
- the pattern of the network can be made up of several metallic substructures 18 (see FIG. 3) electromagnetically coupled together. This coupling strengthens the local electromagnetic field associated with plasmon resonance and therefore the sensitivity of the detection. This coupling will be all the stronger the closer the substructures are. It will also be stronger for substructures whose studs have elliptical sections aligned along their major axis, thanks to the very intense fields created by peak effect in the vicinity of the small terminal radius of curvature of the major axis.
- the pattern of the network may be composed of three coupled elliptical section nanoplots 18, aligned along their major axis, major axis 65 nm and minor axis 40 nm, spaced a few nm apart, without limitation.
- Other objects, spherical fluorescent particles (quantum dots or latex spheres doped with organic dyes for example) or fluorescent molecules 19, playing the role of markers, can also be fixed on the species and thus make it possible to increase the sensitivity of the detection of the modification of the plasmon resonance of the plots when this is close to the wavelength for which the absorption of the particles or molecules 19 is maximum.
- the disturbance of the plasmon resonance of the stud is stronger in the presence of an absorbent species than in the presence of a non-absorbent species.
- the detection of the optical signal on the scale of nanometric plots, that is to say on a sub-wavelength scale is preferably carried out using a confocal microscope if the distance between plots is greater than the micrometer (below, the confocal undergoes the diffraction limit) and using a near field optical microscope of the SNOM type (for “Scanning Near Field Optical Microscope”) if the distance between pads is less than a micrometer and as not limiting in a probe configuration without opening.
- a metallic tip of the SNOM with probe without opening 21 under special lighting conditions can generate a tip effect enhancement of the electromagnetic field in the vicinity thereof, thereby increasing the light intensity near the nano-objects to be detected.
- This point also allows, by coupling its plasmon resonance, if the material constituting this point allows it, with that of the metal nano-stud 12 and possibly that of a metal marker 14, resonance shifts in wavelength still more marked of the system constituted by the point, the pad and the marker, therefore a better sensitivity of the optical detection on the sub-wavelength scale.
- the signal-to-noise ratio of the detection of the near field signal can be improved by vibrating the probe vertically above the sample.
- a SNOM probe is used without opening (as shown in FIG. 3).
- a very small metallic or fluorescent particle 20 typically a few nm
- This particle when it is fluorescent may be, without limitation a molecule or a fluorescent quantum dot, and when it is metallic a sphere of gold or silver of a few nm in diameter.
- This metallic particle 20 has, under optical excitation 15, optical resonances linked to the excitation of localized surface plasmons. This results in the vicinity of the particle 20 under the influence of the species 13 to be detected and to characterize a modification of the plasmon resonance of the particle 20 highlighted by the detection system of the SNOM.
- the enhancement of the electromagnetic field in the vicinity of the particle 20 can be reinforced by a coupling between the plasmon resonances of the particle 20, those of the cylindrical studs with a section in the shape of a circle or ellipse 12 or 18, possibly the resonances of the markers 14 and guided electromagnetic mode 17.
- the presence of biochemical species modifies the intensity and the lifetime of fluorescence of this particle.
- the characteristics of the fluorescence radiation of the particle 20 are modified.
- the sensitivity of the fluorescence detection can be enhanced by the presence of the pads 16 if the plasmon resonance wavelength of these pads is adjusted to the wavelength of absorption or fluorescence emission of the particle 20.
- the fluorescent particle 20 can also be used to reinforce the modification of the plasmon resonance of the pads 16 induced by the species 13. It should be noted that l he invention can be used in a liquid medium, that is to say if the chemical or biological species are in a solution.
Abstract
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Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
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JP2006530462A JP5000300B2 (ja) | 2003-10-09 | 2004-10-07 | 化学種又は生物種用の表面プラズモンマイクロセンサ及びナノセンサ |
EP04805737A EP1670951A2 (fr) | 2003-10-09 | 2004-10-07 | MICRO-CAPTEURS ET NANO-CAPTEURS D’ESPECES CHIMIQUES ET BIOLOGIQUES A PLASMONS DE SURFACE |
US10/574,177 US7705989B2 (en) | 2003-10-09 | 2004-10-07 | Microsensors and nanosensors for chemical and biological species with surface plasmons |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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FR0350663A FR2860872A1 (fr) | 2003-10-09 | 2003-10-09 | Micro-capteurs et nano-capteurs d'especes chimiques et biologiques a plasmons de surface |
FR0350663 | 2003-10-09 |
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WO2005033335A2 true WO2005033335A2 (fr) | 2005-04-14 |
WO2005033335A3 WO2005033335A3 (fr) | 2005-06-02 |
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PCT/FR2004/050494 WO2005033335A2 (fr) | 2003-10-09 | 2004-10-07 | Micro-capteurs et nano-capteurs d’especes chimiques et biologiques a plasmons de surface |
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US (1) | US7705989B2 (fr) |
EP (1) | EP1670951A2 (fr) |
JP (1) | JP5000300B2 (fr) |
FR (1) | FR2860872A1 (fr) |
WO (1) | WO2005033335A2 (fr) |
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Also Published As
Publication number | Publication date |
---|---|
US20070115474A1 (en) | 2007-05-24 |
US7705989B2 (en) | 2010-04-27 |
FR2860872A1 (fr) | 2005-04-15 |
JP2007508536A (ja) | 2007-04-05 |
WO2005033335A3 (fr) | 2005-06-02 |
EP1670951A2 (fr) | 2006-06-21 |
JP5000300B2 (ja) | 2012-08-15 |
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