WO2014167322A1 - Détecteur plasmonique - Google Patents

Détecteur plasmonique Download PDF

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Publication number
WO2014167322A1
WO2014167322A1 PCT/GB2014/051104 GB2014051104W WO2014167322A1 WO 2014167322 A1 WO2014167322 A1 WO 2014167322A1 GB 2014051104 W GB2014051104 W GB 2014051104W WO 2014167322 A1 WO2014167322 A1 WO 2014167322A1
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WO
WIPO (PCT)
Prior art keywords
sample
substrate
detection window
nanostructure
target molecule
Prior art date
Application number
PCT/GB2014/051104
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English (en)
Inventor
Cameron Alexander FRAYLING
Bruno Flavio Nogueira de Sousa SOARES
Original Assignee
Base4 Innovation Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Base4 Innovation Ltd filed Critical Base4 Innovation Ltd
Publication of WO2014167322A1 publication Critical patent/WO2014167322A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • 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
    • G01N21/62—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
    • 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
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G01N21/05—Flow-through cuvettes

Definitions

  • This invention relates to an improved plasmonic device suitable for detecting and identifying small quantities of a target molecule in a gaseous or liquid sample.
  • the device is especially suitable for detecting trace amounts of noxious substances such as biohazards, toxic chemicals, poisonous gases, narcotics or traces of explosive material or residues.
  • Detectors having many different designs have been developed over the years and some are readily available on the market. Those available include devices in which the method of detecting the noxious target is based on chromatography or some form of spectroscopic methods for example fluorescence spectroscopy, infrared spectroscopy, Raman spectroscopy and mass spectrometry. However, in many instance these devices have relatively high detection-limits meaning that, in order to reliably detect trace contaminants, a considerable volume of sample has to be analysed over a significant period of time. And in doing so, the risk of a false reading becomes more likely.
  • US 20110279817 discloses an optical device and associated analysing apparatus for detecting the presence of a target molecule in a gaseous sample.
  • a gas containing the target is caused to flow over a detector comprising a striated dielectric substrate having metal film elements deposited on its ridges.
  • the substrate is illuminated with incident light and Raman-scattered light, emitted by the target absorbed onto the substrate, detected.
  • the benefit of using a striated substrate is that surface plasmon polaritons can be induced in the metal film elements causing the Raman emissions of the target to be enhanced.
  • no provisions are made for driving the target onto the detector from the sample.
  • a similar passive device is taught in US 20120162640 where the substrate comprises an array of nanoparticles arranged on a substrate whose surface is provided with two sets of ridges perpendicular to each other.
  • localised surface plasmons are induced in the various metal nanoparticles to enhance Raman-scattering of the target.
  • US 2009273779 also describes an optical device for detecting a foreign object in a sample using conventional Raman or surface-enhanced Raman spectroscopy.
  • the device is characterised by the presence of a Raman-active substrate consisting of a plasmonic band structure region which can be coupled to optical radiation, the plasmonic band structure region comprising a layer of a first material patterned with an array of sub-regions of a second material, the first material having a first refractive index and the second material having a second refractive index, a side- wall of each sub-region being coated with a metallic layer, wherein the array of sub-regions give rise to a plasmonic band structure, and each sub-region is configured to confine at least one plasmon resonance excited by optical radiation coupled into the plasmonic band structure region.
  • the substrate comprises a regular tiled array in which the sub-regions are located at the vertices of nano-voids.
  • Klarite ® we are aware that such substrates are available commercially under the trade name Klarite ® . Again no provision is made to drive the foreign object to the platform.
  • WO 2007/011876 teaches an apparatus comprising a metal film and at least one resonance configuration formed therein.
  • the configuration itself comprises a pore extending through the film and a single non-annular feature that causes a variation in a dielectric function along a first surface proximate to the aperture.
  • This feature may be a second aperture, protrusion or a depression.
  • the apparatus however is not designed to detect hazardous materials; rather it is orientated towards solving a different technical problem in a completely different technical field; achieving sub-Rayleigh criterion resolution in optical microscopy.
  • EP 1650550 describes a surface plasmon detector in which the analyte (here present in solution) is driven by electrophoresis between a pair of electrodes to a detection zone.
  • the detection zone comprises one of the electrodes attached to the reverse side of which is a prism through which a beam of irradiating light is caused to pass.
  • the device further comprises a photodetector for detecting the intensity of light which has been reflected back out of the prism as a function of various angles of incidence around resonance conditions.
  • the electrode itself is neither nanoporous nor provided with nanostructures. A similar device is taught in JP 1078393.
  • US 2007/0252982 describes a SERS analyser including a tunable resonant cavity in a substrate comprising reflective members and an electro-optic material disposed therebetween. Coupled to the cavity is a Raman signal-enhancing structure. However this device is also not provided with a gradient means for driving the analyte to the cavity/structure arrangement.
  • WO 2010/066727, WO 03/027619, WO 2013/060989, US 2009/045351, JP 2005/337771, GB 2419940, US 2011/166405 and WO 2006/118337 all of which are further illustrative of detectors which exhibit improved sensitivity by virtue of the presence of nanostructures capable of undergoing enhanced localised surface plasmon resonance.
  • US 8129676 describes a device for detecting ions in an analyte in which a stream of ions generated in a separate ionisation stage is directed towards a detector by an electric field.
  • the detector comprises a surface-enhanced Raman spectroscopy system comprised of an array of detector elements each comprising one or more metallic segments separated by an insulator.
  • the patent gives no information about the dimensions of these segments, in particular whether they are nanostructures, or indeed whether they are intentionally stimulated to undergo plasmon resonance at an optimum frequency.
  • a device for detecting the presence of a target molecule in a sample characterised in that it comprises:
  • each nanostructure being capable of generating an associated detection window comprising an electromagnetic field by having induced therein localised surface plasmons
  • the detection device of the present invention can be used to detect the presence of charged and polarisable target molecules present in both gaseous and liquid samples; especially target molecules having a characteristic spectroscopic fingerprint which is significantly different from the other major constituents of the sample.
  • the device Whilst the device can in principle be used for any detection duty, the device is especially useful for the detection of biological pathogens or poisons in drinking water, cooling fluids or air; the detection of residual pesticides in aqueous extracts taken from soils and foodstuffs; the identification of bacterial pathogens in liquids swabbed or taken from human beings, hospital equipment or the like and the detection of narcotics, explosives or their residua on hard surfaces, clothing or the like.
  • the essentially non-perforated substrate employed in the device of the invention is suitably made of a dielectric material such as glass, silicon or silicon nitride and in one embodiment is fabricated from a sheet of such material.
  • a dielectric material such as glass, silicon or silicon nitride
  • the term 'essentially non-perforated' is used to mean that, whilst the substrate may be provided with structures such as wells, channels, cavities and the like, themselves of nano-dimensions, these structures either do not extend completely therethrough or, to the extent they do, they have, at at least one critical point, a narrowing which renders translocation of the target molecule through the substrate impossible.
  • the substrate further comprises a metal or semiconductor layer attached to the surface of the substrate opposite to that bearing the nanostructures (hereinafter referred to as the 'operative surface').
  • the structures referred to above may completely penetrate the dielectric material to the layer providing that they do not completely perforate it.
  • each nanostructure should have a maximum dimension of greater than 1 micron, preferably from 1 to 500 microns, most preferably 1 to 150 microns.
  • these nanostructures may be organised as a plurality of pairs, preferably a regular disposition of such pairs on the operative surface, in which at least some of the nanostructures are spaced apart from their pair by greater than 10 nanometres preferably in the range 10 to lOOnm most preferably in the range 10 to 30nm.
  • the space between the two nanostructures can be made subject to a strong induced electromagnetic field which causes enhancement of any spectroscopic emissions from target molecules present therein.
  • the 'detection window' the space which this electromagnetic field occupies.
  • local 'hotspots' on the operative surface comprising a relatively high density of the nanostructure pairs and hence detection windows, are employed to facilitate the driving of the target molecule generally towards the detection window.
  • the nanostructures themselves are typically fabricated from metals or dielectric materials coated with metals.
  • Metals which can be employed are those capable of undergoing plasmon resonance to a significant extent, for example, gold, silver, copper, aluminium, platinum, palladium, molybdenum and chromium and alloys thereof.
  • the metal used will be gold, silver, copper or an alloy thereof.
  • the nanostructure may have attached to its surface binding sites which are specifically adapted to capture the particular target molecule and enhance the characteristic emissions being sought.
  • Such reactive groups can work by any chemical or physical means; for example when detecting say genetic material characteristic of a bacterial pathogen, e.g. the Legionella bacterium, the reactive group can comprise a polynucleotide probe adapted to bind to certain unique base pair sequences in the bacterium by hybridisation.
  • the detection window can be generated by any suitable arrangement of nanostructures on the operative surface.
  • One simple embodiment comprises a regular disposition of the nanostructures on an otherwise substantially smooth dielectric surface.
  • Such an arrangement has the advantage that it is easy to make; for example by first coating the substrate with a metallic film, then masking up the product and finally etching away the remaining exposed metal with a chemical or ion beam to leave the discrete nanostructures. Methods for carrying out such a method are well known in the art. However more complex structures can be fabricated, for example the nanoparticle array shown in US 20120162640.
  • pairs of triangular nanostructures are employed in a 'bow tie' configuration to reduce the size of the detection window and increase the density of the electromagnetic field generated from a given degree of plasmon resonance.
  • the two nanostructures can be two half annuli juxtaposed to create a 'doughnut' configuration around a substantially circular orifice giving access to the substrate. This configuration is especially useful where the orifice is coincident with a well in the substrate immediately below.
  • the device comprises one or more substrates and their associated nanostructures arranged in a chamber for containing it and the sample.
  • the chamber is provide with a means, such as a pump, fan or the like for passing the sample over the substrate and nanostructures; preferably in a parallel or substantially parallel direction.
  • the chamber may contain different substrates arranged at different locations. This arrangement can advantageously be used to detect different target molecules in a sample having a complex composition. It is especially useful when the means for establishing the gradient is used to selectively direct different target molecules of differing masses to different substrates or regions of the same substrate.
  • the device is further provided with a means to establish a gradient between the sample and the substrate.
  • a means to establish a gradient between the sample and the substrate An effect of this is that the target is thereby induced to flow along the gradient from the sample bulk to the detection window(s) on the substrate.
  • This active, as opposed to passive, transfer of the target molecule to the detection window(s) significantly improves the capture efficiency and therefore the sensitivity of the device. This is very important where the levels of target molecule in the sample are very small and it has the further advantages that (a) the time to a meaningful result is considerably shortened and (b) the signal to noise ratio of the output signal from the device is improved.
  • the gradient established in the device can include for example gradients caused by variations in osmotic pressure, magnetic field or by using chemical reactions to generate a thermodynamic or entropic gradient.
  • the gradient is created by the electrical field produced by applying a potential difference between the sample bulk and a position on the substrate in close proximity to the detection window.
  • the target molecule is then caused to move towards the detection window by electrophoresis, in the case of charged target molecules, or dielectrophoresis, in the case of uncharged, polarisable species.
  • the direction of this electrical field can be reversed; for example to remove the target molecule from the operative surface after detection so that in effect the substrate can be cleaned.
  • the means for establishing this potential difference and the detector are connected via a feedback loop so that, once the detector detects a threshold level of the target molecule, the direction of the field can be varied with time to hold the target molecule in the detection window.
  • the potential difference referred to above can be achieved by locating a first electrode in the chamber, locating a second electrode, of opposite polarity, on the substrate between the nanostructures and then applying a voltage therebetween.
  • this configuration can be modified so that at least part of the substrate region in close proximity or between the nanostructure(s) comprises the types of well, channel or cavity mentioned above.
  • the second electrode sits in or at the bottom of the well, cavity or channel in order to cause the target molecule to pass through the detection window towards its ultimate destination.
  • the side of the substrate opposite to the operative surface is coated with a layer of metal or semiconductor which is able to act as the second electrode.
  • the layer comprises the bottom of or is substantially adjacent to the bottom of the well, channel or cavity referred to above and is made by perforating or etching the substrate towards the internal surface of the layer.
  • the metal or semiconductor layer can be bonded to a previously nano-perforated dielectric substrate precursor.
  • the metal or semiconductor layer can be brought into close proximity with the operative surface by means of wells, cavities or channels of nano-dimensions arranged on a different, suitably opposite, surface of the substrate.
  • the different surface is first patterned with the required wells, cavities and channels and then coated or filled with the metal or semiconductor using known deposition methods.
  • the remainder of the device comprises a high-intensity source of incident electromagnetic radiation, for example a laser, a detector for detecting the characteristic electromagnetic radiation emitted by those target molecules in the detection window and any necessary ancillary optics and electrical circuitry.
  • a high-intensity source of incident electromagnetic radiation for example a laser
  • a detector for detecting the characteristic electromagnetic radiation emitted by those target molecules in the detection window and any necessary ancillary optics and electrical circuitry.
  • Either or both of the source of incident electromagnetic radiation and the detector can be continuously associated with some or all of the nanostructures and the detection windows.
  • the nanostructures and the detection windows can be scanned using for example a movable microscope or raster arrangement.
  • the electromagnetic radiation detected by the detector can in principle be any spectroscopic emission which is characteristic of the target molecule when it is present in the detection window as opposed to a spectral emission or shifts characteristic of the nanostructures themselves.
  • this spectroscopic emission is either fluorescence radiation or Raman-scattered radiation.
  • fluorescence emissions it will often be useful to chemically or biologically label (as the case may be) the target molecule with a dye.
  • labelling will not normally be required and rather wavelengths characteristic of one or more vibrational modes of the target will be monitored by the detector in order to generate a signal characteristic of the wavelength being observed or of a ratio of such wavelengths.
  • Raman spectroscopy the scattering can be derived from single- or multi-photon events.
  • the nature of the detector employed in the device is not critical and can suitably comprise foe example a photodetector, a single photon avalanche diode, an electron-multiplying charge- coupled device or a complementary metal oxide semiconductor device.
  • the detector can additionally be attached to a microprocessor, PC and/or an alarm to process the signal derived therefrom.
  • the whole device can be of unitary design it can also advantageously consist of two components for example a permanent housing in which the source of incident electromagnetic radiation, the detector and optics are located and a disposable cartridge comprising a shell containing the chamber, substrate and nanostructures linked together by microfluidic pathways.
  • the shell then can be made of for example glass, silicon or plastic.
  • the device is of unitary design and is used for guard duties it is preferred that it is provided with a wireless transmitter to send the signal from the detector back to a central location.
  • the device can thus be positioned remotely in which case it may employ a stand-alone power source such as battery if so desired.
  • a method for using the device of the present invention to detect a sample molecule in a sample is characterised by the steps of (a) providing an essentially non- perforated substrate comprising a surface having at least one nanostructure; said nanostructure being adapted to generate an associated detection window comprising an electromagnetic field by having induced therein localised surface plasmons; (b) bringing the sample into contact with the surface whilst maintaining a gradient between the sample and the surface so as to drive the target molecule from the sample into the detection window and (c) detecting electromagnetic radiation characteristic of and emitted by those target molecules in the detection window.
  • the field gradient is preferably an electrophoretic or dielectrophoretic field gradient as described above and the electromagnetic radiation detected is either fluorescence or Raman- scattered radiation.
  • Figure 1 shows a cartridge containing a detection device according to the invention
  • Figure 2 shows a sectional view of the device
  • FIG. 3 illustrates various arrangements of substrate, nanostructures and electrodes suitable for use in the device.
  • a cartridge 1 embodying the detection device is fabricated from polydimethylsiloxane and comprises sample reservoir 2 (to be filled by the user) and an associated valve 4; a reservoir 3 for containing a flushing fluid such as water (suitably prefilled at the source of manufacture) and associated valve 5; a detection device according to the invention 7 and a reservoir for waste 9. 1 further comprises various ancillary valves and pneumatic inlets (labelled v and p respectively) to enable the sample, flushing fluid and effluent to be pumped around the system by peristalsis.
  • ancillary valves and pneumatic inlets labelled v and p respectively
  • 7 and its associated microfluidic pipework is flushed out by opening 5 and pumping the fluid around the system. 5 is then closed and the sample contained in 2 is pumped through 7 and microfluidic lines 6 and 8 into 9 after opening 4.
  • 9 can be isolated by means of valve 10 and emptied via valve 11 and by pumping if so desired. Thereafter 9 can be flushed out by 3 via valve 12.
  • FIG 7 is provided with a transparent glass window 10 in its upper face through which the assembly 13 can be observed.
  • 13 comprises a plinth 14 on which a dielectric sheet 15 is mounted.
  • the exposed surface of 15 is covered with a regular array of gold nanostructures 16 in a bow-tie configuration (see Figures 3A and 3B).
  • an electrode 17 connected to a power source 18.
  • 18 is also connected to an annular electrode 19 of opposite polarity located within chamber 20.
  • the target molecules 21 in the sample are driven by the potential difference between 17 and 19 into the detection window between the jaws of the bow-tie. 16 are continuously illuminated through 10 by laser 22 and its associated optics.
  • Raman-scattered radiation emitted by 20 and passing back out through 10 and via dichroic mirror 23 is detected by photodetector 24 tuned to at least one characteristic Stokes frequency of the target molecule being sought.
  • a signal generated by 24 is then passed to a microprocessor (not shown) for analysis.
  • Figures 3A and 3B illustrate in plan and section an array of bow-tie nanostructures that can be used in the device shown in Figure 2.
  • Figures 3C to 3E illustrate alternative designs in which respectively the electrode 17 sits in a well of nano-dimensions 25; the dielectric sheet 15 is perforated and wherein 17 comprises a layer of gold and wherein 17 extends into wells of nano-dimensions 26 fabricated in the surface of 15 opposite that bearing the nanostructures 16.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
  • Nanotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Molecular Biology (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne un dispositif (7) permettant de détecter la présence d'une molécule cible dans un échantillon, caractérisé en ce qu'il comporte : • une chambre pour contenir l'échantillon ; • un substrat sensiblement non perforé (15) situé dans la chambre ; • au moins une nanostructure (16), juxtaposée sur une surface du substrat (15), chaque nanostructure (16) étant en mesure de générer une fenêtre de détection associée comportant un champ électromagnétique en ayant, induits dans celui-ci, des plasmons de surface localisés ; • une source de rayonnement électromagnétique incident (22) à des fins d'induction desdits plasmons de surface localisés dans ladite (lesdites) nanostructure(s) (16) ; • au moins un moyen (17, 18, 19) permettant d'établir un gradient entre l'échantillon et le substrat (15) pour entraîner la molécule cible de l'échantillon jusque dans la fenêtre de détection et • un détecteur (24) permettant de détecter une caractéristique de rayonnement électromagnétique des, et qu'émettent ces, molécules cibles dans la fenêtre de détection. De préférence, le gradient est un gradient de champ électrophorétique ou diélectrophorétique. Le dispositif (7) convient particulièrement pour la détection de quantités en traces de substances nocives, tels des produits chimiques toxiques, des gaz toxiques, des stupéfiants ou des traces de matières explosives ou de résidus explosifs.
PCT/GB2014/051104 2013-04-09 2014-04-09 Détecteur plasmonique WO2014167322A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB1306447.2 2013-04-09
GB201306447A GB201306447D0 (en) 2013-04-09 2013-04-09 Plasmonic detector

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009030953A1 (fr) * 2007-09-04 2009-03-12 Base4 Innovation Limited Appareil et méthode
US20090279085A1 (en) * 2005-06-14 2009-11-12 Ebstein Steven M Laser-processed substrate for molecular diagnostics
EP2196796A1 (fr) * 2008-12-09 2010-06-16 Imec Spectroscopie optique à molécule unique dans des nanopores semi-conducteurs dans une approche basée sur la transmission
JP2010243223A (ja) * 2009-04-02 2010-10-28 Hitachi High-Technologies Corp 核酸分析デバイス、及び核酸分析装置
US20110053794A1 (en) * 2009-08-26 2011-03-03 Guigen Zhang Nanostructured substrates for surface enhanced raman spectroscopy (sers) and detection of biological and chemical analytes by electrical double layer (edl) capacitance

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090279085A1 (en) * 2005-06-14 2009-11-12 Ebstein Steven M Laser-processed substrate for molecular diagnostics
WO2009030953A1 (fr) * 2007-09-04 2009-03-12 Base4 Innovation Limited Appareil et méthode
EP2196796A1 (fr) * 2008-12-09 2010-06-16 Imec Spectroscopie optique à molécule unique dans des nanopores semi-conducteurs dans une approche basée sur la transmission
JP2010243223A (ja) * 2009-04-02 2010-10-28 Hitachi High-Technologies Corp 核酸分析デバイス、及び核酸分析装置
US20110053794A1 (en) * 2009-08-26 2011-03-03 Guigen Zhang Nanostructured substrates for surface enhanced raman spectroscopy (sers) and detection of biological and chemical analytes by electrical double layer (edl) capacitance

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