CN102628985A - Optical micro-imaging device and method for imaging nanometer surface layer by using super-diffraction off-axis illuminating technology - Google Patents
Optical micro-imaging device and method for imaging nanometer surface layer by using super-diffraction off-axis illuminating technology Download PDFInfo
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Abstract
The invention discloses an optical micro-imaging device and method for imaging a nanometer surface layer by using a super-diffraction off-axis illuminating technology. The device comprises a transparent substrate layer, a nanometer structure layer, a PMMA (Polymethyl Methacrylate) filling layer and a metal/medium multilayer film layer, wherein the nanometer structure layer can be used for modulating the space frequency and polarization direction of illuminating light entering from the back face of a transparent substrate; the metal/medium multilayer film layer can be used for performing high-pass filtering on the space frequency of the modulated illuminating light; and a passing light field of which the local region is 5-200 nanometers can be formed on the upper surface of the metal/medium multilayer film layer. The passing light field can be used for illuminating a sample to be detected by using an ordinary optical microscope, so that surface layer imaging in the depth range of 5-200 nanometers of the sample to be detected is realized. Due to the adoption of an illuminating way provided by the device, the interference of scattered light of the internal structure of the sample on surface layer imaging can be reduced effectively, the resolution of surface layer imaging is increased, and an effective means is provided for the observation and component analysis of the surface layer structure of a substance.
Description
Technical field
The invention belongs to the micro-imaging field, relate to a kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology.
Background technology
The optical microphotograph imaging technique is a kind of effective means of observation microworld, and its efficient, harmless advantage makes it in each field, especially has a wide range of applications in biology, medical science, investigation of materials field.Yet; What ordinary optical microscope used is a light source; When the observation sample surface topography, the sample interior structure also can be illuminated and be produced diffraction or scattering, and this will have a strong impact on the image quality of microscope to sample surface structure (like cell membrane, protein molecule wall etc.).
In order to overcome the shortcoming of ordinary optical microscope to sample top layer image blur; Laser Scanning Confocal Microscope utilizes spot light; The method that point probe is collected has successfully been eliminated the multiple scattered light that when surveying sample, produces, and has realized the blur-free imaging of sample optics transversal section.Yet the improvement of this image quality utilization reduces microscopic fields of view and obtains, though can compensate through scan mechanism, this will cause its complex structure, shortcomings such as inefficiency.
If light source only throws light on to the top layer of sample, and does not get into sample interior, then can avoid of the influence of sample interior scattered light to the top layer structure imaging, improve the top layer image quality.According to this thought; The researchist proposes traditional microslide is replaced with a prism; And make illumination light that total reflection take place in the prism bottom surface, and utilize the disappearance ripple that produces to be thrown light in the sample top layer, can observe top layer imaging more clearly through ordinary optical microscope.Yet the disappearance ripple in this method determines in the penetration depth on sample top layer refractive index and the illumination light incident angle by prism jointly.Because the restriction of prism material, the I of disappearance ripple penetration depth reaches about 200nm, and this still can exist scattering to a certain degree to disturb for observation degree of depth 200nm with interior surface structure.Therefore, in order to observe degree of depth 200nm even thinner surface structure information clearly, need to make up the littler lighting source of a kind of top layer penetration depth.
Summary of the invention
The technical matters that the present invention will solve is: to the problem of ordinary optical microscope to sample top layer image blur; A kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology is proposed; Utilize this method that the top layer illumination of the 200nm and the following degree of depth can be provided, be used to improve the quality of the top layer optical microphotograph imaging of this yardstick.
The technical solution adopted for the present invention to solve the technical problems is: a kind of nanometer top layer optical microphotograph image device that utilizes ultra diffraction off-axis illumination technology, and this device comprises from top to bottom successively:
One transparent substrates layer;
One nanostructured layers is used to contrast the nanostructured that Mingguang City carries out spatial frequency and polarization direction modulation;
One PMMA (polymethylmethacrylate, Polymethylmethacrylate is called for short PMMA) packed layer is used for the PMMA packed layer of leveling nanostructured surface;
One medium/metal stratified film is used for the different space frequency harmonic wave is carried out the medium/metal multilayer film of high-pass filtering.
Preferably, the size of said nanostructured layers and spacing are not more than the illumination light wavelength; The material of said nanostructured layers is a light-proof material, can be metal, also can be medium; The shape of said nanostructured layers can be the solid or the irregular arbitrary face type of rule; The distribution of said nanostructured layers can be uniformly or heterogeneous, can be one dimension or two dimension.
Preferably, the thickness of the relative nanostructured layers upper surface of the upper surface of said PMMA packed layer is 10～30nm.
Preferably, the metal in the medium/metal stratified film can be good conductor materials such as gold, silver, aluminium, and medium can be SiO
2, Al
2O
3Deng the low-consumption optical film material; Said medium/metal stratified film is that medium/metal replaces rete, and each thicknesses of layers scope is 10nm～30nm, and each tunic is thick can be equated, also can be unequal.According to illumination light intensity needs, 2 layers of total numbers of plies of medium/metal multilayer film or multilayer more.
In addition; The present invention provides a kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology; Illumination light is from the transparent substrates layer back surface incident of above-mentioned device; Through nanostructured layers to the modulation of its spatial frequency and polarization direction after entering medium/metal stratified film carry out High frequency filter; Finally the upper surface at the medium/metal stratified film forms the disappearance light field of local in 5～200nm depth range, can be carried out analyzing and testing by optical microscope or spectrometer by the zone, testing sample top layer of disappearance light field illumination.
Preferably, said illumination light can be broad spectrum light source or the LASER Light Source of ultraviolet to visible-range.
Preferably, change the size and the spacing of said nanostructured layers, can regulate the spatial frequency and the intensity distributions of multi-layer film surface disappearance light field.
Preferably, when the material of said nanostructured layers was metal, the distribution that changes nanostructured layers can be regulated the polarization direction and the space distribution of multi-layer film surface disappearance light field.
Preferably, change metal material and the dielectric material in the said medium/metal stratified film or change the ratio of metallic diaphragm and the thickness of media coating, the cutoff frequency in the time of can adjusting high-pass filtering.
Preferably; Change metal material and dielectric material in the said medium/metal stratified film; Perhaps change ratio, the number of plies of metallic diaphragm and the thickness of media coating; Cutoff frequency in the time of can adjusting high-pass filtering, thus the depth of interaction of multi-layer film surface disappearance light field on the sample top layer, range of adjustment 5nm～200nm regulated.
Preferably; Said testing sample by the illumination of disappearance light field can directly utilize optical microscope to observe; Or through excited sample fluorescence; After color filter filters, observe by optical microscope again, or, after color filter filters, carry out analyzing and testing by Raman spectrometer again through excited sample stokes scattering light.
Principle of the present invention is:
When a branch of light is incident to nanostructured from backside of substrate, its amplitude will receive the modulation of nanostructured, and be inspired the harmonic components of various different space frequency.If the material of said nanostructured is a metal; And be the arranged evenly of certain rule; Then will in metal grating, drive the electron production electric current along the electric field component on the grating orientation in the illumination light; Collision makes the lead heating to electronics with the lead lattice atoms again, finally makes electric field energy change into heat energy and loss.Therefore the electric field component along grating orientation can not pass through grating basically, and only the electric field component perpendicular to grating orientation could pass through grating, thereby can realize the modulation of polarized light.Incident light after ovennodulation has entering the medium/metal multilayer film of especial dispersion relation; Under visible light wave range; Said metal has real part and is negative dielectric coefficient, and it will constitute a kind of anisotropy EFFECTIVE MEDIUM jointly with the medium with positive dielectric coefficient.Can know according to EFFECTIVE MEDIUM THEORY; Select suitable metal, dielectric material can make EFFECTIVE MEDIUM have negative and positive dielectric coefficient respectively in tangential and normal direction; Again by dispersion equation
can the equivalence dielectric material dispersion relation be double curve, as shown in Figure 2.Obviously, this equivalence dielectric material is to spatial frequency k
xHave the effect of high-pass filtering, that is to say, have only spatial frequency k
xHigher harmonic components could get into testing sample through the medium/metal multilayer film.It should be noted that the harmonic components that gets into testing sample all has
According to dispersion relation
Can know the normal direction wave vector k of harmonic components in the testing sample
zBe an imaginary number, this shows that the harmonic components that gets into testing sample can only be with the form local of the disappearance ripple upper surface at the medium/metal multilayer film, only therefore can realize the illumination to the testing sample lower surface.In addition; The penetration depth of disappearance ripple is determined by formula
; Material and thickness through changing in the medium/metal multilayer film can make disappearance ripple penetration depth adjusted in 5nm～200nm scope, to satisfy the illumination to testing sample different table layer depth.Illuminated sample can carry out different observation and measurement according to the purpose of illumination light wavelength and analyzing and testing.If illumination light is a visible light, then illuminated sample can directly carry out the top layer micro-imaging by optical microscope; If illumination light is a ultraviolet light, then testing sample should be fluorescent samples or scribbles the sample of fluorescent dye, through to after the fluorescence excitation of sample, the filter plate filtering, carries the fluorescence of sample surface structure information and is observed by optical microscope; If not only need carry out the top layer imaging, but also hope the material in the sample top layer is carried out evaluation and validation, then can also utilize Raman spectrometer that the stokes scattering light that the sample top layer excites is analyzed and monitored to sample.
The present invention compared with prior art has the following advantages:
1, the device that is used for the optical microphotograph imaging of nanoscale top layer among the present invention can provide and thrown light in the top layer in sample 5nm～200nm depth range; Reduced of the influence of sample interior structural scattering light to a greater extent, further improved the quality of sample top layer imaging the top layer image quality.The illumination degree of depth that this device provides, polarization direction can be regulated through size, distribution and the arrangement of nanostructured, and for design proposal provides the more freedom degree, the scope of application is wider.
2, in addition, that this device has is simple in structure, efficient is high, low cost and other advantages, for high resolving power top layer optical microphotograph imaging technique provides novel a, effective method.
Description of drawings
Fig. 1 is the device architecture synoptic diagram that is used for the optical microphotograph imaging of nanoscale top layer according to the invention; Among the figure: 1 is transparent substrates; 2 is nanostructured; 3 is the PMMA packed layer; 4 is metallic diaphragm; 5 is media coating; 6 is testing sample; 7 is cover glass; 8 is filter plate; 9 is optical microscope;
Fig. 2 is the dispersion relation synoptic diagram of medium/metal multilayer film EFFECTIVE MEDIUM among the present invention;
Fig. 3 is for realizing incident light is carried out the one dimension periodic nano-structure synoptic diagram of linear polarization modulation, the direction of shaking thoroughly of the said nanostructured of right side double arrowed line represents among the embodiment 1;
Fig. 4 carries out the alternately two-dimensional and periodic nanostructured synoptic diagram of modulation of polarization direction, the direction of shaking thoroughly of the said nanostructured of right side double arrowed line represents for realizing among the embodiment 2 to incident light;
Fig. 5 be for can realizing incident light is carried out the two-dimensional and periodic nanostructured synoptic diagram of tangential polarization modulation among the embodiment 3, and the annulus that the right side has an arrow is represented the direction of shaking thoroughly of said nanostructured;
Fig. 6 is Ag/Al among the embodiment 1
2O
3Multilayer film is the X-Y scheme of optical transfer function under the 436nm illumination light at wavelength;
Fig. 7 is Ag/Al among the embodiment 1
2O
3The disappearance ripple normalization light intensity of multi-layer film surface is with the change curve that gets into the testing sample degree of depth;
Fig. 8 is Au/SiO among the embodiment 2
2Multilayer film is the X-Y scheme of optical transfer function under the 589nm illumination light at wavelength;
Fig. 9 is Au/SiO among the embodiment 2
2The disappearance ripple normalization light intensity of multi-layer film surface is with the change curve that gets into the testing sample degree of depth.
Embodiment
Introduce the present invention in detail below in conjunction with accompanying drawing and embodiment.But following embodiment only limits to explain the present invention, and protection scope of the present invention should comprise the full content of claim, and promptly can realize the full content of claim of the present invention to the technician in field through following examples.
Embodiments of the invention 1 are to produce the line polarisation, and are used for the device of 15nm yardstick top layer optical microphotograph imaging.Fig. 1 is the device architecture synoptic diagram, and said device is followed successively by from down to up: the suprasil substrate; Chromium sodium rice grating is used to contrast Mingguang City and carries out spatial frequency and polarization direction modulation; The PMMA packed layer is used for the leveling nanostructured surface; Ag/Al
2O
3Multilayer film is used for the different space frequency harmonic wave is carried out high-pass filtering.Chromium sodium rice grating cycle 150nm live width 75nm, the thickness of the relative nanostructured upper surface of the upper surface of PMMA packed layer is 10nm, the dielectric coefficient of Ag is-5.4939+0.2156i Al
2O
3Dielectric coefficient be 3.1, Ag/Al
2O
3The thick 10nm that is of every tunic of multilayer film, totally 64 layers.Illumination light is a partially coherent light, and wavelength is 436nm.After the modulation of chromium nanostructured; Have only the higher harmonic components of horizontal wave vector could be in the incident field, thereby realization be only promptly thrown light on apart from the sample top layer in the multi-layer film surface 15nm scope to the penetration depth of disappearance glistening light of waves field through multilayer film and can only be in the about 15nm scope in the surface of this device multilayer film with the form local of disappearance ripple.This can reduce the interference of the scattered light of sample interior structure to the imaging of sample top layer.
Fig. 3 is the structural representation of chromium nanometer grating in the present embodiment, and the chromium nanometer grating can be used as a wire-grid polarizers, realizes the modulation of illumination light polarization direction, and the four-headed arrow line segment is represented the direction of shaking thoroughly of wire-grid polarizers among the figure.Be to obtain the harmonic components of higher-spatial frequencies, get the chromium grating cycle and be 150nm, live width and be 75nm, highly be 50nm.
Fig. 6 is Ag/Al in this enforcement
2O
3The X-Y scheme of optical transfer function when multilayer film is 436nm in illumination wavelengths.As can be seen from the figure, has only horizontal wave vector k in the incident field after the modulation of chromium nanostructured
x＞2.8k
0Harmonic components could pass through Ag/Al
2O
3Multilayer film can only be with the form local of the disappearance ripple surface at this device multilayer film and satisfy the harmonic components of this requirement.Suppose that the biological sample dielectric coefficient is 2.2, then can obtain disappearing ripple normalization light intensity and the change curve that gets into the sample degree of depth, as shown in Figure 7.Usually cancelling the glistening light of waves overdamp of dying is that initial light intensity 1/e place is its penetration depth, promptly
When getting k
x=2.8k
0The time, the penetration depth of disappearance glistening light of waves field is about 15nm.Testing sample is placed the upper surface of multilayer film, can realize the line polarizing illumination is carried out in the dark zone of sample lower surface 15nm.Owing to adopt the purple light illumination, sample is necessary for fluorescent material or handles through fluorescent dye, after fluorescence excitation, filter plate filtering, carries the fluorescence of the surface structure information in the sample 15nm degree of depth and can be observed by optical microscope.
Embodiments of the invention 2 are to produce tangential polarization light, and are used for the device of 50nm yardstick top layer optical microphotograph imaging.Illumination light is a partially coherent light, and wavelength is 589nm, chromium sodium rice structure, PMMA packed layer and Au/SiO that this device is followed successively by substrate of glass from down to up, radially distributes
2Multilayer film.The thickness of the relative nanostructured upper surface of the upper surface of PMMA packed layer is 10nm, and the dielectric coefficient of Au is-0.9969+0.2319i SiO
2Dielectric coefficient be 2.25, Au/SiO
2The thick 20nm that is of every tunic of multilayer film, totally 32 layers.Fig. 4 and Fig. 5 are the distribution schematic diagram of chromium nanostructured in the present embodiment; Obviously, chromium sodium rice structure radially distributes, according to the principle of work of wire-grid polarizers; The distribution of this metal Nano structure will produce polarized light tangentially, and the direction of shaking thoroughly is shown in the circle that has arrow among the figure.
Fig. 8 is Au/SiO in this enforcement
2The X-Y scheme of the optical transfer function of multilayer film when illumination wavelengths is 589nm as can be seen from the figure, has only horizontal wave vector k in the incident field after the modulation of chromium nanostructured
x＞1.75k
0Harmonic components could pass through Au/SiO
2Multilayer film.Suppose that still the biological sample dielectric coefficient is 2.2, the glistening light of waves that can obtain disappearing is strong and get into the change curve of the sample degree of depth, and is as shown in Figure 9, and the penetration depth of the glistening light of waves field that disappears this moment is about 50nm.Owing to adopt the gold-tinted illumination, therefore can directly utilize optical microscope that the surface structure information of carrying in the sample 50nm degree of depth is observed.
The part that the present invention does not set forth in detail belongs to techniques well known.
Claims (10)
1. one kind is utilized the technological nanometer top layer optical microphotograph image device of ultra diffraction off-axis illumination, and it is characterized in that: this device comprises from top to bottom successively:
One transparent substrates layer;
One nanostructured layers is used to contrast the nanostructured that Mingguang City carries out spatial frequency and polarization direction modulation;
One PMMA packed layer is used for the leveling nanostructured surface;
One medium/metal stratified film is used for the different space frequency harmonic wave is carried out high-pass filtering.
2. the nanometer top layer optical microphotograph image device of the ultra diffraction off-axis illumination technology of utilization according to claim 1, it is characterized in that: the size of said nanostructured layers and spacing are not more than the illumination light wavelength; The material of said nanostructured layers is the metal or the medium of light-proof material; The shape of said nanostructured layers can be the solid or the irregular arbitrary face type of rule; The distribution of said nanostructured can be uniform or heterogeneous, also can be one dimension or the two dimension.
3. the nanometer top layer optical microphotograph image device of the ultra diffraction off-axis illumination technology of utilization according to claim 1, it is characterized in that: the thickness of the relative nanostructured layers upper surface of the upper surface of said PMMA packed layer is 10～30nm.
4. the nanometer top layer optical microphotograph image device of the ultra diffraction off-axis illumination technology of utilization according to claim 1; It is characterized in that: the metal in the said medium/metal multilayer film can be the gold, silver or the aluminium of good conductor material, and medium can be the SiO of low-consumption optical film material
2Or Al
2O
3Said medium/metal multilayer film is that medium/metal replaces rete, and each thicknesses of layers scope is 10nm～30nm, and each tunic is thick can be equated, also can be unequal; According to illumination light intensity needs, 2 layers of total numbers of plies of medium/metal multilayer film or multilayer more.
5. one kind is utilized the technological nanometer top layer optical microphotograph formation method of ultra diffraction off-axis illumination; It is characterized in that: the illumination light accessory rights requires the transparent substrates layer back surface incident of each said device in 1 to 4; Through nanostructured layers to the modulation of its spatial frequency and polarization direction after entering medium/metal stratified film carry out High frequency filter; Finally the upper surface at the medium/metal stratified film forms the disappearance light field of local in 5nm～200nm depth range, can be carried out analyzing and testing by optical microscope or spectrometer by the zone, testing sample top layer of disappearance light field illumination.
6. a kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology according to claim 5 is characterized in that: said illumination light can be broad spectrum light source or the LASER Light Source of ultraviolet to visible-range.
7. a kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology according to claim 5; It is characterized in that: change the size and the spacing of said nanostructured layers, can regulate the spatial frequency and the intensity distributions of multi-layer film surface disappearance light field.
8. a kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology according to claim 5; It is characterized in that: when the material of said nanostructured layers was metal, the distribution that changes nanostructured layers can be regulated the polarization direction and the space distribution of multi-layer film surface disappearance light field.
9. a kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology according to claim 5; It is characterized in that: change metal material and dielectric material in the said medium/metal stratified film; Perhaps change ratio, the number of plies of metallic diaphragm and the thickness of media coating; Cutoff frequency in the time of can adjusting high-pass filtering, thus the depth of interaction of multi-layer film surface disappearance light field on the sample top layer, range of adjustment 5nm～200nm regulated.
10. a kind of nanometer top layer optical microphotograph formation method that utilizes ultra diffraction off-axis illumination technology according to claim 5; It is characterized in that: said by the testing sample of disappearance light field illumination; Can directly utilize optical microscope to carry out the sample observation of nanoscale top layer, or, after color filter filters, observe by optical microscope again through excited sample top layer fluorescence; Or,, color filter carries out analyzing and testing by Raman spectrometer again after filtering through excited sample stokes scattering light.
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| CN102628985B (en) | 2014-06-18 |
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