METHOD AND DEVICE FOR ANALYSING A SAMPLE USING A RESONANT SUPPORT, ILLUMINATED BY INFRARED RADIATION
20220317031 · 2022-10-06
Assignee
- Centre National De La Recherche Scientifique (Paris, FR)
- Cpe Lyon Formation Continue Et Recherche (Villeurbanne, FR)
- ECOLE CENTRALE DE LYON (Ecully, FR)
- INSTITUT NATIONAL DES SCIENCES APPLIQUEES DE LYON (Villeurbanne, FR)
- Universite Claude Bernard Lyon 1 (Villeurbanne, FR)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (Paris, FR)
Inventors
- Mathieu DUPOY (Grenoble Cedex 9, FR)
- Maryse FOURNIER (Grenoble Cedex 9, FR)
- Taha BENYATTOU (Vourles, FR)
- Cécile JAMOIS (Caluire, FR)
- Lotfi BERGUIGA (Lyon, FR)
- Nicolas GAIGNEBET (La Tour d'Aigues, FR)
- Thomas GEHIN (Villeurbanne, FR)
Cpc classification
G01N2021/3196
PHYSICS
International classification
Abstract
A method for analysing a sample uses a resonant support. The sample extends on the support having a surface on which a plurality of separated photonic crystals extend. The sample extends between a light source and the crystals, wherein a resonance wavelength is associated with each crystal addressing the analyte and the wavelengths of the crystals define a resonance spectral band extending between 2 μm and 20 μm. The transmission or reflection of light by each crystal addressing the analyte is maximum at the associated resonance wavelength. The method includes illuminating the support by the light source, the light source emitting an illumination lightwave defining an illumination spectral band which at least partially covers the resonance spectral band, such that a plurality of crystals are simultaneously illuminated; acquiring an image of the support, and then determining the presence of the analyte in the sample from the image.
Claims
1-25. (canceled)
26. A method for analyzing whether a sample includes an analyte, the sample lying on a resonant holder, the resonant holder comprising a surface on which lie a plurality of photonic crystals, which are separate from one another, and the sample being disposed between a light source and the photonic crystals, wherein: one resonant wavelength is associated with each photonic crystal, the resonant wavelengths of the photonic crystals defining a resonance spectral band lying between 2 μm and 20 μm; and transmission or reflection of light by each photonic crystal is maximum at the an associated resonant wavelength; the method comprising: a) illuminating the resonant holder with the light source, the light source emitting an illuminating light wave that defines an illumination spectral band that at least partially covers the resonance spectral band, wherein a plurality of photonic crystals are simultaneously illuminated; b) acquiring, with an image sensor, a measurement image of the resonant holder, the measurement image comprising different regions of interest, each region of interest being optically coupled to one photonic crystal such that each region of interest represents an intensity transmitted or reflected by each photonic crystal, while the resonant holder is illuminated; and then determining whether the analyte is present in the sample on the basis of the measurement image; wherein each photonic crystal comprises: first holes formed in a thin layer and having a first dimension, the first dimension being a radius or a diagonal, the first holes defining a first periodic pattern; second holes formed in the thin layer and having a second dimension, the second dimension being strictly smaller than the first dimension, the second holes defining a second periodic pattern; the second pattern and the first pattern are offset with respect to each other, parallel to the surface of the resonant holder, by a spatial offset, the spatial offset being variable between at least two different photonic crystals; and the resonant wavelength associated with each photonic crystal depends on the first dimension, on the second dimension, and on the spatial offset.
27. The method of claim 26, further comprising: c) using a reference image, the reference image being representative of an image acquired by the image sensor when the resonant holder is illuminated in the illumination spectral band in a reference configuration, in an absence of analyte; d) comparing the measurement image, acquired in b), with the reference image, in c); and e) based on the comparison, determining whether the analyte is present in the sample.
28. The method of claim 27, comprising, prior to a), forming the reference image, comprising: illuminating the resonant holder with the light source, in the illumination spectral band, without analyte between the light source and the resonant holder; and acquiring an image of the resonant holder with the image sensor, the image thus acquired forming the reference image.
29. The method of claim 27, comprising obtaining the reference image by: illuminating, in the illumination spectral band, a reference holder that is considered to be representative of the resonant holder illuminated in a), an amount of analyte between the reference holder and the light source being considered to be zero; and forming an image of the reference holder, the image thus formed corresponding to the reference image.
30. The method of claim 27, wherein the resonant holder comprises reference photonic crystals the resonance spectral band of which is one of: not modified by the analyte, and corresponds to a wavelength not absorbed by the sample; and wherein the reference image is an image of the reference photonic crystals when they are illuminated in the illumination spectral band.
31. The method of claim 30, wherein the reference image and the measurement image form two parts of the same image acquired by the image sensor.
32. The method of claim 27, wherein: the sample is a liquid; the resonant holder makes contact with the sample; the resonant holder comprises photonic crystals addressing the analyte, which are configured to make contact with the analyte; the measurement image comprises regions of interest that are associated with the analyte and optically coupled to the photonic crystals addressing the analyte; under an effect of contact with the analyte, the resonant wavelength of the photonic crystals addressing the analyte undergoes a spectral variation; and the intensity of the illumination varies, in the resonance spectral band, according to a spectral illumination function.
33. The method of claim 32, wherein, for each photonic crystal addressing the analyte, the resonant wavelength depends on a refractive index of the sample at an interface between the sample and the photonic crystal, the refractive index varying as a function of an amount of analyte making contact with the photonic crystal.
34. The method of claim 32, wherein e) comprises: on a basis of the measurement image, determining a measurement profile of the intensity of the regions of interest associated with the analyte; and on a basis of the reference image, determining a reference profile representative of the intensity in absence of analyte or in a presence of a known amount of analyte in the sample, of the regions of interest associated with the analyte, wherein whether the analyte is present is determined depending on a shift of the measurement profile with respect to the reference profile.
35. The method of claim 34, wherein the amount of analyte is estimated depending on the shift of the measurement profile with respect to the reference profile.
36. The method of claim 32, wherein e) comprises: in the measurement image, determining a measurement position of a region of interest exhibiting a maximum intensity value among the regions of interest associated with the analyte; and in the reference image, determining a reference position of a region of interest exhibiting a maximum intensity value among the regions of interest associated with the analyte, wherein the presence of the analyte is determined depending on a shift of the measurement position with respect to the reference position.
37. The method of claim 36, wherein an amount of analyte is estimated depending on the shift of the measurement position with respect to the reference position.
38. The method of claim 32, wherein e) further comprises estimating an amount of analyte in the sample, depending on the comparison between the measurement image and the reference image.
39. The method of claim 26, wherein the photonic crystals lie in a holder plane, the illuminating light wave propagating to the resonant holder parallel to a propagation axis that is perpendicular or substantially perpendicular to the holder plane.
40. The method of claim 26, wherein the resonant holder is disposed between the light source and the image sensor, such that, in b), each region of interest formed in the measurement image is representative of an intensity transmitted by the photonic crystal to which said region of interest is optically coupled.
41. The method of claim 26, wherein: the resonant holder bounds a half-space comprising the light source; and the image sensor is disposed in the half-space, such that each region of interest formed in the measurement image is representative of an intensity reflected by the photonic crystal to which said region of interest is optically coupled.
42. The method of claim 26, wherein: the sample is disposed between the light source and the resonant holder; the analyte is able to absorb the illuminating light wave, emitted by the light source, in an absorption spectral band; and the absorption spectral band comprises at least one absorption wavelength corresponding to a resonant wavelength of a photonic crystal, the method comprising, on the basis of the measurement image: determining at least one photonic crystal, the resonant wavelength of which corresponds to the absorption wavelength; identifying the resonant wavelength of said photonic crystal; and determining whether the analyte is present on a basis of the resonant wavelength of the photonic crystal thus determined.
43. The method of claim 42, comprising: on a basis of the measurement image, estimating an absorption light intensity, transmitted or reflected by at least one photonic crystal the resonant wavelength of which is comprised in the absorption spectral band; estimating a reference light intensity, transmitted or reflected by said photonic crystal, in the absence of analyte; and estimating an amount of analyte on a basis of the absorption light intensity and of the reference light intensity.
44. The method of claim 26, wherein the sample is one of a gas, liquid and solid.
45. A device for detecting an analyte within a sample, comprising: a light source; an image sensor; and a resonant holder disposed between the light source and the image sensor, wherein the image sensor is configured to acquire an image of the resonant holder, the resonant holder is configured to be placed in contact with a sample, and the resonant holder comprises an area containing photonic crystals, which are separate from one another, wherein: the light source is configured to simultaneously illuminate a plurality of photonic crystals in an illumination spectral band lying between 2 μm and 20 μm; one resonant wavelength is associated with each photonic crystal, wherein transmission or reflection of the light by each photonic crystal is maximum at an associated resonant wavelength; at least two photonic crystals are configured to make contact with the analyte, said at least two photonic crystals being associated with two different resonant wavelengths, the resonant wavelength associated with each photonic crystal depending on an amount of analyte in contact with the photonic crystal; each photonic crystal comprises: first holes formed in a thin layer and have a first dimension, wherein the first dimension is a radius or a diagonal and the first holes define a first pattern; second holes formed in the thin layer and that have a second dimension, wherein the second dimension is strictly smaller than the first dimension and the second holes define a second periodic pattern; and the second pattern and the first pattern are offset with respect to each other, parallel to the surface of the holder, by a spatial offset that is variable between at least two different photonic crystals; and the resonant wavelength associated with each photonic crystal depends on the first dimension, on the second dimension, and on the spatial offset.
46. The device of claim 45, wherein: the photonic crystals are aligned parallel to a longitudinal axis; and the resonant wavelengths respectively associated with two adjacent photonic crystals are offset by a discretization pitch comprised between 1 nm and 50 nm.
47. The device of claim 45, wherein: a plurality of photonic crystals are aligned in a row parallel to a longitudinal axis; and the resonant wavelength respectively associated with a photonic crystal gradually increases, or decreases, along the longitudinal axis.
48. The device of claim 45, wherein: the resonant holder comprises a plurality of rows of photonic crystals parallel to one another, and the photonic crystals form columns, parallel to a lateral axis, such that the photonic crystals of a given column have a same resonant wavelength.
49. The device of claim 45, wherein the photonic crystals addressing the analyte are covered with a functionalization layer configured to selectively capture the analyte on the photonic crystals.
50. The device of claim 45, wherein the resonant holder comprises: photonic crystals of rank 1, which are configured to capture a first analyte; and photonic crystals of rank j, which are configured to capture a jth analyte different from analytes captured by the photonic crystals of rank lower than j.
Description
FIGURES
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DESCRIPTION OF PARTICULAR EMBODIMENTS
[0134]
[0135] By analyte, what is meant is a chemical or biological species the presence, and possibly an amount, of which in the sample it is desired to determine. The analyte 21 may, for example, be a chemical molecule, a protein, a peptide, an antibody, an antigen, a fragment of a nucleotide sequence, or a particle. By particle, what is meant for example is a biological cell, a droplet that is insoluble in a medium, or a nanobead. It may also be a question of a microorganism, a bacterium for example, a yeast or a microalgae. Preferably, a particle has a diameter, or is inscribed in a diameter, smaller than 20 μm, or even than 10 μm or than 5 μm.
[0136] When the sample is a gas, the analyte is a gas species with a spectral band of light absorption Δλ.sub.a comprised in the spectral band of illumination Δλ. The gas species may for example be CH.sub.4, CF.sub.4, CO.sub.2, NO, or NH.sub.3. Generally, the gas species exhibits absorption that is considered to be significant in the spectral band of illumination. The case of a gas sample is addressed in the second embodiment.
[0137] When the sample is a solid, it is sufficiently transparent, in at least one part of the spectral band of illumination Δλ, so as to allow light to propagate through the sample. It may for example be a question of a thin slide of biological tissue obtained from a sample of biopsy or smear type. It is for example a question of a pathology slide. It may also be a question of a dry extract obtained by drying a bodily liquid, of a thin slide of dried blood for example. The analyte may then be a molecule, or one portion of a molecule, or of a molecular bond, exhibiting a spectral band of absorption Δλ.sub.a in the spectral band of illumination Δλ. The case of a solid sample is addressed in the second embodiment.
[0138] By resonant holder, what is meant is a holder one portion of which is able to resonate, so as to transmit or to reflect a maximum light intensity at a resonant wavelength. The structure of the resonant holder is described below.
[0139] The sample 20 may take the form of a drop deposited on the resonant holder 15. It may also be a question of a liquid or gas confined in a fluidic chamber 18 associated with the resonant holder 15. The resonant holder may for example consist of a wall of the fluidic chamber 18 in which the sample lies. The fluidic chamber 18 is preferably transparent in the spectral band of illumination Δλ. It may for example comprise or be made of materials that are transparent in the infrared: it is for example a question of silicon, of germanium, of calcium fluoride (CaF.sub.2) or of barium fluoride (BaF.sub.2), or of zinc sulfide (ZnS) or of a chalcogenide.
[0140] The light source 10 may be monochromatic or polychromatic. The illuminating wave 11 lies in a spectral band of illumination Δλ. The spectral band of illumination lies between 2 μm and 20 μm. Preferably, the spectral band of illumination extends between a minimum wavelength and a maximum wavelength, the minimum wavelength being strictly longer than 1.5 μm, or even 2 μm. Thus, the spectral band of illumination may lie in the near infrared (typically between 2 μm and 3 μm), or in the mid-infrared (typically between 3 and 8 μm), or in the far infrared (typically up to 20 μm).
[0141] The light source 10 may be formed by a heated filament, forming a light source the spectral emission of which is close to that of a black body. The light source may be formed by a laser. It may be a question of a wavelength-tunable laser, and for example of a quantum cascade laser (QCL). The light source may comprise a plurality of elementary QCL sources, emitting in various spectral bands of illumination, respectively.
[0142] In a first embodiment, described with reference to
[0143] Preferably, the light source 10 is placed at a distance A from the resonant holder 15, such that the light wave 11 reaches the latter in the form of a plane wave. A collimating optical element, known to those skilled in the art, may be placed between the light source 10 and the sample 20, so as to form a plane light wave 11.
[0144] The device 1 also comprises an image sensor 30 that is sensitive to the infrared. The image sensor is preferably a pixelated sensor, comprising pixels 31 arranged in a matrix array. The pixels of the image sensor 30 define a detection plane P.sub.30. In this example, the image sensor is formed by a matrix array of bolometers, each bolometer of the matrix array having a spectral band of detection comprised between 2 μm and 20 μm. Each bolometer forms one pixel. In the examples described below, each pixel is formed by a vacuum-encapsulated bolometer.
[0145] Preferably, the detection plane P.sub.30 is placed perpendicular to the propagation axis Z, or substantially perpendicular to the latter. By substantially perpendicular, what is meant is perpendicular to within an angular tolerance of +20°, or preferably ±10°, or even ±5°. Thus, the illuminating wave 11 emitted by the light source 10 reaches the holder at a normal incidence, to within the angular tolerance.
[0146] Preferably, the resonant holder 15 defines a holder plane P.sub.15. The holder plane P.sub.15 lies perpendicular to the propagation axis Z, or substantially perpendicular to the latter. The resonant holder extends along a longitudinal axis X and a lateral axis Y. The axes X and Y are coplanar with the holder plane. They are secant, and preferably perpendicular.
[0147] Preferably, the detection plane P.sub.30 extends parallel to the holder plane P.sub.15.
[0148] An important aspect of the invention is that the resonant holder 15 comprises nanostructured elementary zones that are separate from one another, each elementary zone forming one photonic crystal 16.sub.k. Thus, the resonant holder comprises photonic crystals 16.sub.k, which are different from one another, and which are spaced from one another. The index k is an integer strictly comprised between 1 and K, K corresponding to the number of photonic crystals formed on the resonant holder 15.
[0149] Each photonic crystal 16.sub.k has a resonant wavelength λ.sub.k that is specific thereto. At the resonant wavelength λ.sub.k that is associated with it, each photonic crystal 16.sub.k exhibits a peak in light transmission or reflection. Each crystal thus has a resonant wavelength in reflection and a resonant wavelength in transmission.
[0150] The image sensor 30 is configured such that the pixels 31 are divided into groups of pixels 32.sub.k, the pixels 31 belonging to the same group of pixels 32.sub.k being optically coupled to the same photonic crystal 16.sub.k.
[0151] By group of pixels, what is meant is a pixel or a set of pixels that are adjacent to one another and optically coupled to the same photonic crystal.
[0152] By optically coupled, what is meant is that the pixels 31 of the same group of pixels 32.sub.k collect light the intensity of which is at least 80% or 90% due to light propagating through the photonic crystal 16.sub.k.
[0153] In the example shown in
[0154] Alternatively, as shown in
[0155] The light source is placed at a distance A from the resonant holder, such that a plurality of photonic crystals are simultaneously illuminated by the light source. Thus, at least 3, and preferably at least 5 or 10 photonic crystals are simultaneously illuminated by the light source. The illumination of the resonant holder is preferably extensive, in the sense that the illuminated area is preferably larger than 1 mm.sup.2, or even 1 cm.sup.2. The illuminated area may correspond to the field of observation of the image sensor 30. In the lens-less configuration described with reference to
[0156] The device 1 also comprises a processing unit 40, for example a microprocessor, configured to process and/or display images acquired by the image sensor 30. The processing unit may be connected to a memory 41, comprising instructions for implementing image-processing algorithms. The processing unit 40 is preferably connected to a screen 42.
[0157] The configurations schematically shown in
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[0159] Thus, the thin layer 22 comprises a plurality of resonant photonic crystals 16.sub.k that are separate from one another, and that each have a resonant wavelength λ.sub.k in transmission or in reflection. The thickness of the thin layer 22 is adjusted depending on the spectral band of illumination. It is preferably comprised between 50 nm to a few μm. When the spectral band of illumination Δλ extends beyond 5 μm, the thickness of the thin layer may for example reach 1.5 μm to 3 μm. The holes 23 extend through the thin layer 22, about an axis perpendicular to the plane of the holder. The holes 23 may in particular extend about an axis parallel to the propagation axis Z.
[0160] In the holder plane P.sub.15, the photonic crystals 16.sub.k have a diagonal or a diameter comprised between 10 μm and 500 μm, of 100 μm for example. Each photonic crystal is borne by a transparent or translucent membrane, formed by a layer 24, which may be made of silicon. Fabrication of the photonic crystals is described below, with reference to
[0161] The spectral properties of the photonic crystals with respect to transmission and reflection of light may be determined using simulations performed via computer codes. Specifically, the properties of propagation of light in the photonic crystals stem from their specific periodic arrangement. These propagation properties may easily be modeled, by a person skilled in the art, on the basis of Maxwell's equations. In the remainder of this description, the modelling was carried out using the software package Rsoft and a method of RCWA type.
[0162] By photonic crystal, what is meant is a structure the refractive index of which varies periodically, on the wavelength scale, in one or more directions. In the examples described in this description, the photonic crystals are two-dimensional, this being a preferred configuration. The invention therefore takes advantage of the development of techniques for micro-structuring dielectrics, semiconductors or metals, allowing control of the interaction of electromagnetic waves in three-dimensional structures based on the arrangement of materials of various indices.
[0163] Each photonic crystal 16.sub.k transmits (or reflects) light according to a spectral transmission (or reflection) function describing a variation in a light intensity transmitted (or reflected) by the photonic crystal, as a function of wavelength. The exploitation of Fano resonance makes it possible to design compact resonant photonic crystals, which may be illuminated collectively by one illuminating light wave, at a normal incidence. This makes it possible to simultaneously illuminate, in a simple manner, photonic crystals distributed in one or two dimensions. The spectral transmission function of each photonic crystal exhibits a maximum, at a resonant wavelength in transmission. Analogously, the spectral reflection function exhibits a maximum, at a resonant length in reflection.
[0164] The spectral transmission or reflection function, and in particular the resonant wavelengths in transmission or in reflection, depend first of all on the structure of each photonic crystal, i.e. on the size and on the spatial arrangement of the holes 23 produced in the thin layer 22, to form the photonic crystal 16.sub.k. In reflection or in transmission, the resonant wavelength λ.sub.k also depends on the refractive-index contrast between the thin layer 22 and the sample, in the holes 23. The resonant wavelength also depends on the thickness of the thin layer 22. It also depends on the refractive index and on the thickness of the transparent layer 24.
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[0166] Before it is brought into contact with the sample, the resonant holder 15 will have undergone a surface functionalization, in the holder plane P.sub.15, such that each photonic crystal is able to capture a predetermined analyte. Surface functionalization is a concept known to those skilled in the art. It consists in adding a specific function to the surface to be functionalized, by nanostructuring, or by depositing a coating, or by adsorbing or grafting molecules with specific properties. In the present case, the surface functionalization confers, on each photonic crystal, a property enabling selective capture of an analyte. The analyte may be captured via formation of a covalent, hydrogen or electrostatic bond with the analyte and/or via grafting of the analyte with a ligand placed on the functionalized capturing surface. Following the functionalization, the holder plane P.sub.15 is also a functionalized holder plane, i.e. one that has been functionalized to capture one or more analytes.
[0167] When the resonant holder is placed in contact with the sample, the sample fills the holes 23 of each photonic crystal 16.sub.k. The spectral properties of each photonic crystal 16.sub.k with respect to transmission (or reflection) of the light are then governed by the structure of each photonic crystal (in particular the size and distribution of the holes 23) and the respective refractive indices of the sample 20 and of the thin layer 22.
[0168] In
[0169] Because of the surface functionalization, when the desired analyte is present in the sample, the concentration of analyte captured by the holder increases after the latter has been brought into contact with the sample. This results in a local variation in the index of the sample, at the interface between the sample and the functionalized holder. Under the effect of such a variation in index, the spectral transmission (or reflection) properties of each functionalized photonic crystal change—in particular the resonant wavelength of the photonic crystals changes.
[0170] A noteworthy aspect of the invention is that the photonic crystals are dimensioned such that: [0171] the respective resonant wavelengths of the photonic crystals, and notably of photonic crystals addressing the same analyte, are different from one another; [0172] the spectral offset between two resonant wavelengths of two adjacent photonic crystals is small and controlled.
[0173] All the resonant wavelengths of photonic crystals addressing the same analyte lie between a minimum resonant wavelength λ.sub.r,min and a maximum resonant wavelength λ.sub.r,max. The latter bound a spectral band of resonance Δλ.sub.r. It is necessary for the spectral band of illumination Δλ of the light source to be, at least partially, included in the spectral band of resonance Δλ.sub.r. The spectral band of illumination Δλ may be the same as the spectral band of resonance Δλ.sub.r. It may be wider or narrower than the spectral band of resonance Δλ.sub.r.
[0174] In the spectral band of resonance Δλ.sub.r, which is defined by the photonic crystals of the holder, the intensity of the light wave 11 emitted by the light source 10 is not constant. In the spectral band of resonance Δλ.sub.r, the intensity of the light wave 11 is variable, and follows a spectral illumination function ƒ. The spectral illumination function ƒ defines the intensity of the incident light wave 11 at various wavelengths of the spectral band of resonance, such that:
I(λ)=ƒ(λ)
where: [0175] λ is a wavelength of the spectral band of resonance Δλ.sub.r; [0176] I(λ) is the intensity of the illuminating wave 11, or illumination intensity, at the wavelength λ.
[0177] It is important that the illumination function ƒ not be constant, i.e. that the intensity of the illuminating wave not be uniform in the spectral band of resonance Δλ.sub.r. The illumination function ƒ may thus be monotonic in the spectral band of illumination, and for example an increasing or decreasing function. It may also increase (decrease, respectively) to an extremum and then decrease (increase, respectively) from the extremum. When the light source is monochromatic, the illumination function forms a peak in the spectral band of resonance Δλ.sub.r.
[0178] In the preferred embodiment, two adjacent photonic crystals 16.sub.k, 16.sub.k+1 have resonant wavelengths that are offset by a known spectral offset dλ.sub.k. The spectral resolution of the method is dependent on the spectral offset dλ.sub.k. In absolute value, the spectral offset dλ.sub.k is preferably smaller than 10 nm, and more preferably smaller than nm, or even than 2 nm. It corresponds to a spectral discretization pitch, with which the illuminating light wave 11 is discretized, as described with reference to
[0179] On capture of the analyte, the resonant wavelength of each photonic crystal changes, passing from a value λ.sub.ref,k, in the absence of analyte, to a value λ.sub.k in the presence of analyte, with λ.sub.k=λ.sub.ref,k+δA (1). It will be noted that the variation in resonant wavelength δλ is the same for all the photonic crystals addressing a given analyte. In other words, the sensitivity of the resonant wavelength to variations in index is the same for all the various photonic crystals.
[0180] The change in resonant wavelength δλ is consecutive to the capture of the analyte, which generally results in an increase in the index at the interface between the resonant holder and the sample. The increase in index causes a variation in the resonant wavelength of each photonic crystal.
[0181] Thus, the invention is based on a measurement of a variation in resonant wavelength δA under the effect of capture of the analyte by photonic crystals addressing the same analyte.
[0182] The variation δλ in resonant wavelength is observed by taking into account a reference configuration, in which the captured amount of analyte is known. Preferably, in the reference configuration, the amount of analyte captured by each photonic crystal is zero. This is the case that will be considered in the remainder of the description. Alternatively, provision may be made for an embodiment in which in the reference configuration corresponds to a known captured amount of analyte.
[0183] The precision with which the variation δλ in resonant wavelength is estimated depends on the spectral offset dλ.sub.k between two adjacent photonic crystals. The smaller the spectral offset dλ.sub.k, the better the resolution with which the variation δλ is estimated.
[0184] The variation in resonant wavelength δλ is determined by comparing a reference image I.sub.ref of the resonant holder, in the reference configuration, with an image I of the resonant holder taken after capture of the analyte. The image I of the holder, after capture, is called the measurement image.
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[0187] The regions of interest ROI.sub.k are distinct from one another, and aligned parallel to the photonic crystals 16.sub.k. In
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[0189] Comparison of the measurement image I shown in
[0190] On the basis of the measurement image and of the reference image, it is possible to form an intensity profile representative of a spatial distribution of the intensity of each region of interest ROI.sub.k along the longitudinal axis X.
[0191] On account of the alignment of the photonic crystals along the same axis X, and of the small spectral offset dλ.sub.k between two adjacent photonic crystals, each profile has approximately the same shape as the spectral illumination function. Thus, the resonant holder 15 allows spectral information to be converted into spatial information. It acts in the same way as a spectrometer. The spatial information corresponds to a position, along the axis X, of each region of interest ROI.sub.k. A spectral variation δλ, in the present case the variation in resonant wavelength affecting each photonic crystal, results in a spatial shift Δx of the profile, between the reference image and the measurement image. As the spectral offset dλ.sub.k between two adjacent photonic crystals is set, the variation in resonant wavelength δλ may be estimated, using the expression:
where: [0192] Δk corresponds to the spatial variation, which is obtained by comparing the measurement image and the reference image, and which represents the number of regions of interest 32.sub.k forming the observed spatial variation Δx; [0193] Σ.sub.k.sup.k+Δkdλ.sub.k corresponds to the sum of the spectral offsets between the photonic crystals respectively associated with the regions of interest forming the observed spatial variation Δx.
[0194] Preferably, the spectral offset dλ.sub.k between two adjacent photonic crystals may be considered to be constant and equal to dλ. In this case, the preceding expression becomes:
δλ=Δk×dλ (3)
[0195] The advantage of using a non-uniform spectral illumination function ƒ will now be understood. It facilitates a comparison of the intensity profiles in the reference configuration and in the measurement configuration, respectively, so as to allow the number Δk of regions of interest ROI.sub.k by which the intensity profile shifts between the reference configuration and the measurement configuration to be estimated. In the example shown in
[0196] In contrast to certain prior-art devices that require a spectrometer to be used, the invention transfers the spectral splitting function to the resonant holder 15. This allows the use of a simple image sensor, which is clearly less expensive and complex to implement.
[0197] According to one embodiment, the light source is monochromatic, and may be tunable. The spectral band of illumination may then be narrower than or equal to the spectral resonant-wavelength offset dλ.sub.k between two adjacent photonic crystals. In order to be able to estimate the variation δλ in resonant wavelength under the effect of the capture of the analyte, the spectral band of illumination then corresponds to a resonant wavelength of a single photonic crystal in the reference configuration, i.e. in the absence of analyte capture, and in the measurement configuration.
[0198] The light source may be monochromatic and tunable, so that the spectral band of illumination may be modified.
[0199] The reference image may be an image, of the sample, acquired before the capture. In this case, the reference image is taken at an initial time, at which the capture of the analyte by the holder is considered to be negligible. It may for example be a time close to the time at which the holder and the sample are brought into contact. The measurement image is then acquired after the reference image.
[0200] Alternatively, the reference image is an image obtained with a reference holder that is considered to be representative of the holder brought into contact with the sample. The reference holder is then brought into contact with a reference sample that is considered to be representative of the analyzed sample. The reference image is stored in memory. Such an embodiment assumes a good reproducibility in the fabrication of the holders.
[0201] Thus, the reference image, taken into account to estimate the variation in resonant wavelength, may be: [0202] acquired by the image sensor, prior to the acquisition of the measurement image; [0203] or obtained using another holder, then stored in memory; [0204] or acquired by the image sensor, simultaneously with the measurement image, using the same resonant holder, the reference image corresponding to the image of photonic crystals configured not to capture the analyte. It is a question, for example, of non-functionalized photonic crystals. In this case, one portion of the image acquired by the image sensor forms the measurement image: this corresponds to the regions of interest of the acquired image representing the photonic crystals having captured the analyte, or, more generally, making contact with the analyte. Another portion of the image acquired by the image sensor forms the reference image: it corresponds to the regions of interest of the acquired image representing the photonic crystals not having captured the analyte, or, more generally, not making contact with the analyte. For example, the photonic crystals functionalized to capture the analyte are aligned in a first row, whereas the photonic crystals not configured to capture the analyte are aligned in a second row, the two rows being parallel to each other. The image of the first row forms the measurement image, whereas the image of the second row forms the reference image. Thus, from the same image acquired by the image sensor, the measurement image and the reference image may be obtained.
[0205] Regardless of the embodiment, the reference image is formed by illuminating the holder or the reference holder, in a spectral band of illumination Δλ and according to a spectral illumination function ƒ equal to those employed to acquire the measurement image. In other words, the reference image and measurement image of the employed holder(s) are obtained under the same illumination conditions.
[0206]
[0207] The method makes it possible to detect a presence of the analyte in the sample, this corresponding to the observation of a non-zero variation δλ in resonant wavelength. It also makes it possible to estimate an amount of analyte captured by the resonant holder, with an precision depending on the number of photonic crystals and on the spectral offset dλ.sub.k between the resonant wavelength of two adjacent photonic crystals.
[0208] On the basis of the amount of analyte captured by the holder, it is then possible to estimate an amount of analyte in the sample, said amount possibly for example being expressed in the form of a concentration. A calibration, using samples the analyte concentration of which is known, allows a calibration function, relating the amount of analyte captured by the holder to the analyte concentration initially present in the sample, to be obtained.
[0209]
[0210] Step 100: bringing the sample into contact with the resonant holder.
[0211] Step 110: illuminating the sample in the spectral band of illumination Δλ, according to the spectral illumination function ƒ.
[0212] Step 120: acquiring a measurement image I of the resonant holder.
[0213] Step 130: taking into account of a reference image I.sub.ref, the reference image possibly being an image of the holder acquired just after step 110, prior to the acquisition of the measurement image, this forming the optional step 115.
[0214] Step 140: comparing the measurement image I with the reference image I.sub.ref, so as to estimate a variation in the resonant wavelength δλ of the photonic crystals. It is notably a question of determining the number of photonic crystals (or the number of regions of interest) corresponding to the variation in resonant wavelength δλ. Knowing the spectral offset dλ.sub.k between the resonant wavelengths of the photonic crystals, it is thus possible to determine the variation in resonant wavelength δλ.
[0215] Step 150: on the basis of the comparison, determining whether the analyte addressed by the photonic crystals is present in the sample, and possibly estimating an amount of analyte in the sample.
[0216] In the example shown in
[0217] As described with reference to the first embodiment, the photonic crystals are distributed in columns Y.sub.1, Y.sub.2, Y.sub.k, Y.sub.K, such that the photonic crystals belonging to two adjacent columns Y.sub.k have, in the reference configuration, the same resonant wavelength λ.sub.k,ref.
[0218] This configuration, which is referred to as the two-dimensional configuration, corresponds to the addition of one dimension to the embodiment described with reference to
[0219] It will be noted that the device allows a plurality of photonic crystals to be illuminated simultaneously, this making it possible to obtain exploitable information relating to various analytes from a single image acquired using a simple image sensor.
[0220] According to this embodiment, steps 100 to 150, which were described with reference to
[0221] In the reference image (
[0222] According to one such embodiment, the device allows the presence of various analytes in the sample to be detected simultaneously, and the concentration thereof to be estimated.
[0223]
[0224] The modeling the results of which are shown in
[0227] In this example the first periodic pattern P.sub.1 and the second periodic pattern P.sub.2 define a rectangular mesh, of period a along X and 2a along Y, the rectangular mesh being reproduced in two orthogonal directions of translation. In this example, the two directions of translation correspond to the longitudinal axis X and to the lateral axis Y, respectively. The first periodic pattern P.sub.1 and the second periodic pattern P.sub.2 are interlaced in the sense that, in one direction of translation, in the present case the Y-direction, an alternation is observed between rows of first holes 23.sub.1, parallel to the longitudinal axis X, and rows of second holes 23.sub.2, also parallel to the longitudinal axis X. In this example, the period a.sub.1 (along X) of the first periodic pattern P.sub.1 is equal to the period a.sub.2 (along X) of the second periodic pattern P.sub.2, the common period of the two patterns being denoted a. The value of the period is typically between 100 nm and 5 μm. In this example, the period a is larger than 1 μm. Generally, if A is a wavelength belonging to the spectral band of illumination Δλ, the period a is comprised between λ/4 and λ.
[0228] One advantage of a double-period resonant structure is the ability to adjust a spatial offset δ between the first pattern and the second pattern. The spatial offset δ corresponds to a distance between the respective rows of first and second holes.
[0229] It is believed that such a double-period structure allows high quality factors, typically of the order of 1000, i.e. a full width at half maximum of the peak of 1 nm for a resonant wavelength of 1000 nm, to be obtained.
[0230] The modeled curves in
[0235] The quality factor depends on the ratio R.sub.2/R.sub.1. When R.sub.2/R.sub.1 is of the order of 0.6, the quality factor is equal to a few tens. When R.sub.2/R.sub.1 tends toward 1, the quality factor rapidly increases. The quality factor may then exceed a several thousand.
[0236] Generally speaking, the first radius R.sub.1 is smaller than a/2. The ratio R.sub.2/R.sub.1 is preferably higher than or equal to 0.8. The period a may vary between 100 nm and 5 μm, as indicated above.
[0237]
[0238] Thus, another advantage of the double-period structure such as described above is to allow the resonant wavelength to be adjusted: [0239] by modifying the period a of the patterns, this leading to a relatively large variation in the resonant wavelength, typically 10 nm when the period a is varied by 4 nm; [0240] by changing the spatial offset δ of both patterns, this leading to a smaller variation in the resonant wavelength, typically 2 nm for a spatial offset δ of 16 nm, in the particular example of geometry described with reference to
[0241] The quantitative values given in the previous paragraph were obtained by modeling.
[0242] The inventors have fabricated a resonant holder so as to carry out experimental tests, in a spectral band comprised between 920 nm and 980 nm. The characteristics of the resonant holder were as follows: [0243] thin layer 22: silicon—thickness 58 nm; [0244] diameter D of each photonic crystal: 100 μm; [0245] radius R.sub.1 of the first holes 23.sub.1: a/3; [0246] radius R.sub.2 of the second holes 23.sub.2: 0.9 R.sub.1: [0247] period a: variable between 288 nm and 304 nm, with period increments Aa of 4 nm; [0248] spatial offset of the first pattern with respect to the second pattern: variable between 0 nm, 16 nm, 23 nm, 28 nm, 33 nm and 37 nm. Thus, for a given period a, 6 photonic crystals were able to be obtained, each array respectively having a spatial offset δ chosen from 16 nm, 23 nm, 28 nm, 33 nm and 37 nm.
[0249]
[0253] A resonant wavelength, in reflection, at about 7.8 μm with a quality factor of 1000 may clearly be seen.
[0254]
[0255] The main steps of fabrication of the photonic crystals are described below, with reference to
[0256]
[0257] In order to obtain an upper layer 22 of Si having the desired thickness, said layer undergoes a thermal oxidation, then a chemical etch in hydrofluoric acid to remove the resulting SiO.sub.2 so as to obtain a layer 22 of Si of 58 nm thickness and a layer 22′ of SiO.sub.2 of 80 nm thickness on the upper layer 22. This step is schematically shown in
[0258]
[0259] The steps shown in
[0260] The steps shown in
[0261] In the fabricating process described above, the holes 23 are formed by photolithography then etching, this allowing wafer-level fabrication of photonic crystals on the holder. Such a process makes it possible to simultaneously obtain a high number of photonic crystals, this being advantageous from an economic point of view. Alternatively, the holes may be formed by scanning with an electronic beam (e-beam), this making it possible to obtain more precise geometries, to the detriment of the cost and rate of fabrication.
[0262] Trials have been carried out to characterize the resonant holder 15 thus formed, and more precisely the photonic crystals 16.sub.k. To this end, a drop of water was placed on a resonant holder 15, and a glass slide of 17 μm thickness applied to confine the drop of water. A light-emitting diode centered on 940 nm was used as light source, the spectral band of illumination extending to ±40 nm with respect to the central wavelength of 940 nm. The transmission of the photonic crystals was then characterized via transmission spectrometry tests.
[0263]
[0264] For various fabricated photonic crystals 16.sub.k, other tests, in fact reflectometry tests, made it possible to establish a resonant wavelength.
[0265] Subject to adjustment of the dimensions of the photonic crystals, and in particular of the period a, of the radii R.sub.1 and R.sub.2, and of the thickness of the thin layer 22 in which the photonic crystals are formed, the results described with reference to
[0266] A resonant holder comprising such photonic crystals allows a resonant wavelength to be identified in a relatively wide spectral band, of spectral width typically larger than 50 nm, and in this example of spectral width equal to 60 nm. When the photonic crystals are aligned, as shown in
[0267] In one advantageous configuration (shown in
[0268] Provision may be made to use a cross polarization, combining an upstream polarizer, upstream of the sample, and a downstream polarizer, downstream of the sample, in the embodiments shown in
[0269]
[0270] Because of the variation in index between water and ethanol, the wavelength of each photonic crystal undergoes a spectral variation δλ, in the present case an increase, this resulting in a slight shift, to the left in the image, of the discretized spectral illumination function.
[0271] According to a second embodiment, the ability of the invention to determine, on the basis of a measurement image, an absorption wavelength λ.sub.a of an analyte, is exploited.
[0272] In this embodiment, the sample comprises an analyte with a spectral band of absorption Δλ.sub.a lying in the spectral band of resonance Δλ.sub.r and in the spectral band of illumination Δλ.
[0273] As shown in
[0274] Just as in the first embodiment, the resonant holder 15 comprises various photonic crystals 16.sub.k, each photonic crystal having a resonant wavelength λ.sub.k in transmission. Preferably, as in the first embodiment, the photonic crystals 16.sub.k are aligned in the same row, for example parallel to the longitudinal axis X, such that the resonant wavelength λ.sub.k gradually increases (or decreases) from one photonic crystal 16.sub.k, 16.sub.k+1 to the next, along the longitudinal axis X.
[0275] As shown in
[0276] In this example, the structure of the photonic crystals 16.sub.k is such as described in the first embodiment, and more precisely with reference to
[0277] As previously mentioned, the various resonant wavelengths λ.sub.k of the various photonic crystals define a spectral band of resonance Δλ.sub.r.
[0278] As described above, the sample may be a gas, in which case the analyte is a gas species. The gas may be for example air, or a gas exhaled by a living being, or a gas employed in an industrial process.
[0279] The sample may be a solid or a liquid, in which case the analyte is a molecule. Specifically, it is known that the spectral transmission function of light by a sample varies depending on the composition of the latter, because of the presence of absorption peaks corresponding to the vibrational modes of the molecules from which the sample is made up. The presence of absorption peaks is the basis of vibrational spectrometry methods such as infrared spectroscopy or Raman spectrometry.
[0280] By transmission function tr(λ), what is meant is a ratio between: [0281] an intensity i(λ), of a light wave having propagated through the sample 20, and detected by the image sensor 30, at the wavelength λ; [0282] an intensity, referred to as the reference intensity i.sub.ref(λ), of a light wave detected by the image sensor, at the same wavelength λ, in the absence of sample.
[0283] According to the Beer-Lambert law:
[0284] The absorbance abs(λ), at the wavelength λ, is obtained according to the expression:
[0285] According to this embodiment, the spectral illumination function ƒ may be variable in the spectral band of resonance, or constant in the spectral band of resonance Δλ.sub.r.
[0286] In the examples schematically shown in
[0287]
[0288]
[0289]
[0290]
[0291]
[0292] In the measurement images, the intensity of the regions of interest corresponding to the photonic crystals the resonant wavelength of which is in the spectral band of absorption of an analyte depends on the absorption of the illuminating light wave by the analyte, as expressed by expression (2). The absorption in particular depends on an absorption coefficient of the analyte, said coefficient varying as a function of wavelength, and of the amount of analyte present in the sample. Since the value of the absorption coefficient, in the various spectral bands of resonance, is known, an estimate of the attenuation allows the amount of analyte present in the sample to be estimated.
[0293] The measurement image allows the intensity i (λ.sub.a) of the light wave having propagated through the sample, in the presence of analyte, at an absorption wavelength λ.sub.a, to be estimated. The absorption wavelength is a resonant wavelength λ.sub.r of a photonic crystal belonging to the spectral band of absorption Δλ.sub.a. The reference intensity i.sub.ref(λ.sub.a) of the light wave having propagated through the sample, in the absence of analyte, at the absorption wavelength, may be estimated on the basis of a reference image acquired in the absence of a sample containing the analyte.
[0294] Alternatively, the reference intensity i.sub.ref(λ.sub.a) may be estimated from the image acquired by the image sensor, in the presence of the sample, on the basis of a region of interest ROI.sub.k optically coupled to a photonic crystal 16.sub.k the resonant wavelength λ.sub.0 of which is considered to not be absorbed by the analyte. Knowledge of the illumination function ƒ makes it possible to pass from an intensity measured at the unabsorbed wavelength λ.sub.0, to the reference intensity i.sub.ref(λ.sub.a) at the absorption wavelength λ.sub.a.
[0295] The examples described with reference to
[0296] When the sample is a gas, a pre-concentrating loop may be placed upstream of the device. As known, such a loop comprises an absorbent medium able to reversibly trap one or more gas species flowing through the loop. This allows reversible storage of an analyte. Prior to the measurement, the absorbent media is configured to release the trapped gas species. The use of such a pre-concentrating loop allows the amount of analyte between the light source and the resonant holder of the device to be increased.
[0297] Regardless of the embodiment, the sample is preferably spatially homogeneous. By spatially homogeneous, what is meant is that, at least in one row of photonic crystals, the amount of analyte in the sample is considered to be constant.
[0298] The invention will possibly be used to detect and quantify an analyte in samples, for example in the field of biological analysis or to assist with medical diagnosis. The invention may also be employed to monitor industrial processes, for example in the food industry, or even to monitor the environment.