CN102156115B - Coherent anti-Stokes Raman scattering microscopic method and system of super-diffraction limit - Google Patents
Coherent anti-Stokes Raman scattering microscopic method and system of super-diffraction limit Download PDFInfo
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Abstract
The invention is applicable to the field of laser detection and provides a coherent anti-Stokes Raman scattering microscopic method and system of a super-diffraction limit. In the method, a spatial resolution of the super-diffraction limit can be obtained through increasing an additional detecting light which is collinearly focused on a sample together with a pump light and a Stokes light at the same time, enabling the additional detecting light to exhaust phonons generated by the pump light and the Stokes light on a periphery of a focal spot so as to form useless CARS (Coherent Anti-Stokes Raman Scattering) signals, enabling photons in accordance with a phase matching condition in the delayed detecting light to hit phonons at a central area of the focal spot so as to form useful CARS signals, and separating the useless CARS signals. Because supercontinuum laser with a broad band is adopted as the pump light and the Stokes light, a complete CARS spectrum signal of a molecule is obtained to image the whole molecule.
Description
Technical field
The invention belongs to the laser detection field, relate in particular to a kind of ultra diffraction limit coherent anti-stokes raman scattering microscopic method and system.
Background technology
The spatial resolution of optical microscope receives the restriction of diffraction limit, and finally wavelength and the microscopical numerical aperture by light wave determines.If represent its limiting resolution with δ, λ representes optical wavelength, and NA representes the numerical aperture of optical system, then δ=0.61 λ/NA.In general, δ >=200nm.
In recent years, the diffraction limit resolution of fluorescent microscope is owing to taked special measure to obtain important breakthrough.Two kinds of different measures have mainly been taked; A kind of formation method that is based on simple scan; Adopt stimulated emission to exhaust (Stimulated Emission Depletion; STED) method, the peripheral excited molecule of the point spread function that utilizes ring light that exciting light is formed produces stimulated radiation and is exhausted, and has only the centre of exciting light point spread function to produce fluorescence; And it is just more little to produce the centre of fluorescence with the raising of ring light intensity, thereby can obtain not receive the arbitrarily small nanometer of diffraction limit restriction and even the spatial resolution of Ya Nami.Another kind is based on unimolecule location, wide field (the single molecule localization of fluorescence molecule switching effect; SML) formation method; This method has different mutation; As photosensitive location micro-(photoactivatedlocalization microscopy, PALM), micro-(fluorescence photoactivationlocalization microscopy is FPALM) with micro-(the stochastic opticalreconstruction microscopy of optics reconstruct at random for the photosensitive location of fluorescence; STORM); But its basic functional principle is the same, all is the switching effect that utilizes fluorescence molecule, through regulating sparse the exciting that exciting light and deactivation light intensity ratio realize the mark fluorescent molecule; Through steps such as timesharing imaging, unimolecule location and image synthesize, obtain the spatial resolution image of ultra diffraction limit at last again.Because general molecule does not fluoresce or sends out very weak fluorescence, therefore, above-mentioned fluorescence microscopy all will be carried out fluorescence labeling to sample with the fluorescence molecule of special preparation.
So far the method for utilizing cold method to obtain breakthrough diffraction limit resolution also has two kinds; A kind of is the coherent anti-stokes raman scattering microscope that utilizes an enhancement effect; But it will use most advanced and sophisticated enhancing, can only form images to sample surfaces, uses and receives very big restriction.Another kind method is the excited Raman micro-imaging, and it uses two bundle stokes lights, a branch of Gaussian that is, and another is restrainted ringwise, similar above-mentioned fluorescently-labeled STED method.But this method can only obtain the Raman spectrum of a certain chemical bond of molecule, and depending merely on this, to compose the molecular species of differentiating in the sample be difficult.Obtain other Raman spectrum of molecule, just require laser instrument to have wide tuning range.More serious problem be need be very long test duration could obtain the complete Raman spectrum of molecule.For obtaining the ultra diffraction resolution image of three-dimensional of sample, required imaging time is oversize.
CARS (Coherent Anti-Stokes Raman Scattering) utilizes the third-order non-linear effect of four-wave mixing and the signal that produces; Therefore micro-for traditional CARS, only what in this excitation volume, studied still is the macroscopic appearance of a large amount of molecules to resolution for
of traditional optical diffraction limit resolution.Up to 2009, several kinds of possible technological approaches have just been inquired in the world first about realizing that the ultra diffraction limit of CARS is differentiated.Mainly contain three kinds of schemes; A kind of is to utilize the vibrational energy level realization population inversion of a branch of ring light with point spread function scope inside circumference molecule; After pump light and stokes light incident, then can not strengthen producing desired phonon again through resonance, thereby the point spread function scope is dwindled.Another kind method is the local vibration that the contoured in the point spread function scope circularizes, and makes it identical with the anti-Stokes signal frequency that produces, through interference technique raising spatial resolution.The third method adopts common structural light to improve spatial resolution, but this has not been proper nano-resolution micro-imaging, because surpassed 100nm on the spatial resolution principle of utilizing this method to obtain.At present, still be in the theoretical demonstration stage aspect the research of CARS nano-resolution micro-imaging technique in the world,, also only limiting to single chemical bond imaging molecule even the scheme of introducing above can also seeing simultaneously can realize nano-resolution.
Summary of the invention
The purpose of the embodiment of the invention is to provide a kind of ultra diffraction limit coherent anti-stokes raman scattering microscopic method, is intended to further improve the resolution of existing CARS microscopic method.
The embodiment of the invention is achieved in that a kind of ultra diffraction limit coherent anti-stokes raman scattering microscopic method may further comprise the steps:
Generate ultrashort laser pulse;
Make said ultrashort laser pulse produce first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse, the wavelength of said second ultrashort laser pulse is different with the wavelength of said the 3rd ultrashort laser pulse;
Said first ultrashort laser pulse is converted into super continuous spectrums laser, second ultrashort laser pulse after postponing as surveying light, is shaped as the additional probe light that central light strength is little, ambient light is powerful with said the 3rd ultrashort laser pulse;
Make said additional probe light and said super continuous spectrums laser while conllinear focus on sample; Form useless CARS signal behind the phonon that said super continuous spectrums laser produces and the photon collision of said additional probe light; And the phonon of focal spot periphery exhausted, after colliding, the phonon of photon and focal spot central area that more said additional probe light delay arrives the detection light of sample is formed with the CARS signal of usefulness;
Obtain said useful CARS signal, carry out micro-imaging.
Another purpose of the embodiment of the invention is to provide a kind of ultra diffraction limit coherent anti-stokes raman scattering microscopic system, and said system comprises:
Femto-second laser is used to generate ultrashort laser pulse;
Beam split and non-linear device are used to make said ultrashort laser pulse to produce first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse, and the wavelength of said second ultrashort laser pulse is different with the wavelength of said the 3rd ultrashort laser pulse;
Reforming unit is used for said first ultrashort laser pulse is converted into super continuous spectrums laser;
Apparatus for shaping is used for said the 3rd ultrashort laser pulse is shaped as the additional probe light that central light strength is little, ambient light is powerful;
Deferred mount is used to regulate the time that said second ultrashort laser pulse and additional probe light arrive sample, with second ultrashort laser pulse after postponing as surveying light;
The conllinear focalizer; Be used to make said additional probe light and said super continuous spectrums laser while conllinear to focus on sample; Form useless CARS signal behind the phonon that said super continuous spectrums laser produces and the photon collision of said additional probe light; And the phonon of focal spot periphery exhausted, after colliding, the phonon of photon and focal spot central area that more said additional probe light delay arrives the detection light of sample is formed with the CARS signal of usefulness;
Imaging device is used to obtain said useful CARS signal, carries out micro-imaging.
Embodiment of the invention increase by one and pump light and stokes light while conllinear focus on the additional probe light of sample; This additional probe light exhausts the phonon that pump light and stokes light produce at its focal spot periphery; Form useless CARS signal; The photon that meets phase-matching condition in the detection light that postpones then with after the phonon collision of focal spot central area is formed with the CARS signal of usefulness, with useless CARS Signal Separation, can obtain the spatial resolution of ultra diffraction limit.
Description of drawings
Fig. 1 is the realization flow figure of the ultra diffraction limit coherent anti-stokes raman scattering microscopic method that provides of the embodiment of the invention;
Fig. 2 is the structural drawing of the ultra diffraction limit coherent anti-stokes raman scattering microscopic system that provides of the embodiment of the invention;
Fig. 3 is another structural drawing of the ultra diffraction limit coherent anti-stokes raman scattering microscopic system that provides of the embodiment of the invention;
Fig. 4 is super continuous spectrums laser, detection light and the intensity distributions synoptic diagram of additional probe light on sample;
Fig. 5 is CARS nano-resolution microscope distinguishing analogue result, and wherein solid line is represented
(peak light intensity of additional probe light) is I
DepObtain point spread function when (pump light and stokes light exhaust the required additional probe light light intensity of phonon of its generation when being in peak light intensity) 50 times, full width at half maximum is 41 nanometers; Dotted line is represented the point spread function under the conventional optical microscope diffraction-limited.
Fig. 6 is the relativeness figure that surveys light frequency, additional probe light frequency, useless CARS signal spectral coverage and useful CARS signal spectral coverage.
Embodiment
In order to make the object of the invention, technical scheme and advantage clearer,, the present invention is further elaborated below in conjunction with accompanying drawing and embodiment.Should be appreciated that specific embodiment described herein only in order to explanation the present invention, and be not used in qualification the present invention.
Embodiment of the invention increase by one and pump light and stokes light while conllinear focus on the additional probe light of sample; This additional probe light exhausts the phonon that pump light and stokes light produce at its focal spot periphery; Form useless CARS signal; The photon that meets phase-matching condition in the detection light that postpones then with after the phonon collision of focal spot central area is formed with the CARS signal of usefulness; CARS Signal Separation with useless can obtain the spatial resolution of ultra diffraction limit, thereby obtains the complete CARS spectrum signal of molecule.
The ultra diffraction limit coherent anti-stokes raman scattering microscopic method that the embodiment of the invention provides may further comprise the steps:
Generate ultrashort laser pulse;
Make said ultrashort laser pulse produce first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse, the wavelength of said second ultrashort laser pulse is different with the wavelength of said the 3rd ultrashort laser pulse;
Said first ultrashort laser pulse is converted into super continuous spectrums laser, second ultrashort laser pulse after postponing as surveying light, is shaped as the additional probe light that central light strength is little, ambient light is powerful with said the 3rd ultrashort laser pulse;
Make said additional probe light and said super continuous spectrums laser while conllinear focus on sample; Form useless CARS signal behind the phonon that said super continuous spectrums laser produces and the photon collision of said additional probe light; And the phonon of focal spot periphery exhausted, after colliding, the phonon of photon and focal spot central area that more said additional probe light delay arrives the detection light of sample is formed with the CARS signal of usefulness;
Obtain said useful CARS signal, carry out micro-imaging.
The ultra diffraction limit coherent anti-stokes raman scattering microscopic system that the embodiment of the invention provides comprises:
Femto-second laser is used to generate ultrashort laser pulse;
Beam split and non-linear device are used to make said ultrashort laser pulse to produce first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse, and the wavelength of said second ultrashort laser pulse is different with the wavelength of said the 3rd ultrashort laser pulse;
Reforming unit is used for said first ultrashort laser pulse is converted into super continuous spectrums laser;
Apparatus for shaping is used for said the 3rd ultrashort laser pulse is shaped as the additional probe light that central light strength is little, ambient light is powerful;
Deferred mount is used to regulate the time that said second ultrashort laser pulse and additional probe light arrive sample, with second ultrashort laser pulse after postponing as surveying light;
The conllinear focalizer; Be used to make said additional probe light and said super continuous spectrums laser while conllinear to focus on sample; Form useless CARS signal behind the phonon that said super continuous spectrums laser produces and the photon collision of said additional probe light; And the phonon of focal spot periphery exhausted, after colliding, the phonon of photon and focal spot central area that more said additional probe light delay arrives the detection light of sample is formed with the CARS signal of usefulness;
Imaging device is used to obtain said useful CARS signal, carries out micro-imaging.
Below in conjunction with specific embodiment realization of the present invention is described in detail.
Fig. 1 shows the realization flow of the ultra diffraction limit coherent anti-stokes raman scattering microscopic method that the embodiment of the invention provides, and details are as follows.
In step S101, generate ultrashort laser pulse;
The embodiment of the invention adopts femto-second laser 1 to generate ultrashort laser pulse 10.
In step S102, make above-mentioned ultrashort laser pulse produce first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse, the wavelength of this second ultrashort laser pulse is different with the wavelength of the 3rd ultrashort laser pulse;
The embodiment of the invention has number of ways makes ultrashort laser pulse 10 produce first ultrashort laser pulse 11, second ultrashort laser pulse 12 and the 3rd ultrashort laser pulse 13.
For example, second ultrashort laser pulse 12 and the 3rd ultrashort laser pulse 13 produce through nonlinear effect by ultrashort laser pulse 10.Further, first ultrashort laser pulse 11, second ultrashort laser pulse 12 and the 3rd ultrashort laser pulse 13 all can be produced by ultrashort laser pulse 10 pump light parametric oscillators 2.Particularly, (OpticalParameter Oscillator, OPO), thereby acquisition has the ultrashort laser pulse output of the different wave length of wide tunable range to the ultrashort laser pulse 10 pump light parametric oscillators 2 that femto-second laser 1 generates, and is as shown in Figure 2.Through the constant ultrashort laser pulse of the wavelength of optical parametric oscillator 2 output as first ultrashort laser pulse 11; Through the ideler frequency light of optical parametric oscillator 2 output as second ultrashort laser pulse 12; As the 3rd ultrashort laser pulse 13, wherein the centre wavelength of ideler frequency light is grown to few 300nm than the centre wavelength of flashlight through the flashlight of optical parametric oscillator 2 output.
For another example, only the 3rd ultrashort laser pulse 13 by ultrashort laser pulse 10 beam split after nonlinear effect produce.Particularly; The ultrashort laser pulse 10 that femto-second laser 1 generates is divided into three beams through the first optical splitter BS1 and the second optical splitter BS2; Wherein a branch of ultrashort laser pulse after frequency-doubling crystal 3 frequencys multiplication as the 3rd ultrashort laser pulse 13; Other two bundle ultrashort laser pulses are respectively as first ultrashort laser pulse 11 and second ultrashort laser pulse 12, and are as shown in Figure 3.Present embodiment makes the centre wavelength of second ultrashort laser pulse 12 grow to few 300nm than the centre wavelength of the 3rd ultrashort laser pulse 13 through frequency multiplication.
In step S103, first ultrashort laser pulse is converted into super continuous spectrums laser, second ultrashort laser pulse after postponing as surveying light, is shaped as the additional probe light that central light strength is little, ambient light is powerful with the 3rd ultrashort laser pulse;
The embodiment of the invention utilizes first ultrashort laser pulse, 11 pump photon crystal optical fibres (PCF) 4 to obtain super continuous spectrums laser 14; This super continuous spectrums laser 14 comprises pump light and the stokes light that is used to produce phonon, and wherein pump light and Stokes light intensity all are Gaussian distribution.Simultaneously, the 3rd ultrashort laser pulse 13 is shaped as the additional probe light 15 that central light strength is little, ambient light is powerful through phase place zone plate 5, this additional probe light 15 (ω
P ' 2) light intensity distribute ringwise.In addition, as surveying light 16, this surveys light 16 (ω with second ultrashort laser pulse 12 after postponing
P ' 1) intensity be Gaussian distribution.
As preferably, each spectral power fluctuation is not more than 50% in the super continuous spectrums laser 14, and each spectral component is continuous and spectral bandwidth is 10～500 nanometers, and the time range at each spectral component place is not more than 1 psec.Through control lag device 61,62, make and survey light 16 than additional probe light 15 delays 0.5～1 psec arrival sample 7; It is short that the centre wavelength of additional probe light 15 is surveyed the centre wavelength of light 16, therebetween at a distance from the bandwidth that is not less than useful CARS signal; The bandwidth of surveying light 16 is not more than 5 nanometers.
In step S104; Make additional probe light and super continuous spectrums laser while conllinear focus on sample; This additional probe light combines to form useless CARS signal with the phonon that super continuous spectrums laser produces; And the phonon of focal spot periphery exhausted, arrive the detection light of sample and the CARS signal that the phonon at focal spot center combines to be formed with usefulness than the additional probe light delay;
As long as the spectral difference between pump light and the stokes light is consistent with the molecular vibration at focal spot place spectrum, and satisfies energy and phase-matching condition, coherence resonance will take place, produce corresponding phonon.Because annular additional probe light has depletion action to pump light and stokes light at the phonon that the focal spot periphery produces; Finally make pump light, stokes light and the detection light useful CARS signal that acting in conjunction produces in the focal spot center much smaller than diffraction limit; And along with the enhancing of additional probe light, spatial resolution also can further be improved.
In the embodiment of the invention, super continuous spectrums laser, additional probe light and detection light three's focal spot center is overlapping.Fig. 4 shows super continuous spectrums laser, detection light and additional probe light along the radially intensity of (r) (I) variation of focal spot.The spatial resolution (point spread function full width at half maximum) of utilizing the quantum optical theory to be derived does
Wherein
And I
DepPeak light intensity and pump light and the stokes light of representing additional probe light respectively exhausts the required additional probe light light intensity of phonon of its generation when being in peak light intensity.Therefrom can find out; When not adding the additional probe light time; Top expression formula then provides the limit space resolution of diffraction-limited under the nonlinear optical effect; It when not considering nonlinear optical effect diffraction limit determined value will be improved
doubly, this is consistent with the result that common nonlinear optics microscope obtains.When additional probe light
During increasing, then spatial resolution can obtain further improvement, for example
Be I
Dep50 times the time, then spatial resolution can reach 41 nanometers, and is as shown in Figure 5, far surpasses the spatial resolution that diffraction limit can reach.Say on the principle; As long as
constantly increases, spatial resolution just can constantly be improved.
The embodiment of the invention makes additional probe light 15 and super continuous spectrums laser 14 focus on sample 7 via 9 while of microcobjective conllinear through regulating first deferred mount 61 with light combination mirror 8; This moment, 14 line focuses of super continuous spectrums laser formed a focal spot, and this focal spot acts on sample 7 and produces phonon.Because the light intensity of additional probe light 15 distributes ringwise, therefore the conllinear photon that focuses on the additional probe light 15 of sample 7 forms useless CARS signal and exhausts the peripheral phonon of focal spot with the phonon collision simultaneously.Then, regulate second deferred mount 62 and make detection light 16 postpone the arrival of 0.5～1 psec, but conllinear focuses on sample 7 than additional probe light 15.The phonon collision of the photon of this detection light 16 and focal spot central area is formed with the CARS signal of usefulness, and is as shown in Figure 6.
The four bundle laser (pump light, stokes light, detection light and additional probe light) that adopt because of the embodiment of the invention are ultrashort laser pulse; Their duration is much smaller than the dephasing time of molecular vibration; Can use time-resolved method fully (is that intensity is that the additional probe light and the super continuous spectrums laser of annular spread arrives sample simultaneously; And intensity is that the detection light of Gaussian distribution is with respect to the additional probe light delay), eliminate the off-resonance ground unrest.Therefore, as long as the time spectrum structure of super continuous spectrums laser is more suitable for the microscopical requirement of CARS, not only can obtain far to surpass the spatial resolution of diffraction limit, and can obtain the complete CARS spectrum signal of broadband, ultra broadband even molecule of high s/n ratio through scanning.Therefore, the embodiment of the invention can be measured the molecular species that the nano-resolution scope contains, and can estimate the ratio that various molecules are shared.
In step S105, obtain useful CARS signal, carry out micro-imaging.
With off-resonance ground unrest and useless CARS target signal filter, the spectrometer 22 via band EMCCD (Electron-Multiplying CCD) detector forms images the embodiment of the invention again through band pass filter 21.
Because the useless CARS signal that the focal spot periphery produces has the spectral band that separates fully with the useful CARS signal that the focal spot central area produces, and can the two be separated through band pass filter.Contracted through above-mentioned additional probe optical pressure and to have produced the volume of useful CARS signal; Not only can obtain nano level spatial discrimination ability; And because detection light has the time delay less than 1 psec with respect to super continuous spectrums laser; Thereby also suppressed the off-resonance ground unrest simultaneously, obtained the complete CARS spectrum signal of molecule with high spectral resolution.On this basis, through nanometer micrometric displacement platform mobile example or high precision vibration mirror scanning, then can obtain the CARS three-dimensional nano-resolution micro-image of biological sample different molecular to be measured.
For example, the nanometer displacement that focuses on scanning or the sample of focal spots through above-mentioned four bundle laser (pump light, stokes light, detection light and additional probe light) obtains the spatial resolution image of the ultra diffraction limit of three-dimensional of any degree of depth of sample.
Fig. 2 and Fig. 3 show the structural principle of the ultra diffraction limit coherent anti-stokes raman scattering microscopic system that the embodiment of the invention provides, and for the ease of describing, only show the part relevant with the embodiment of the invention.
Femto-second laser 1 is used to generate ultrashort laser pulse 10.
Beam split and non-linear device are used to make ultrashort laser pulse 10 to produce first ultrashort laser pulse 11, second ultrashort laser pulse 12 and the 3rd ultrashort laser pulse 13, and wherein the wavelength of second ultrashort laser pulse 12 is different with the wavelength of the 3rd ultrashort laser pulse 13.
Reforming unit is used for first ultrashort laser pulse 11 is converted into super continuous spectrums laser 14.This reforming unit has one and supplies the photonic crystal fiber (PCF) 4 of first ultrashort laser pulse, 11 pumpings.
Apparatus for shaping 5 is used for the 3rd ultrashort laser pulse 13 is shaped as the additional probe light 15 that central light strength is little, ambient light is powerful.
The conllinear focalizer is used to make additional probe light 15 and 14 while of super continuous spectrums laser conllinear to focus on sample 7; Form useless CARS signal behind the phonon that super continuous spectrums laser 14 produces and the photon collision of additional probe light 15; And the phonon of focal spot periphery exhausted, after colliding, the phonon of photon and focal spot central area that postpones to arrive the detection light 16 of samples 7 than additional probe light 15 is formed with the CARS signal of usefulness.
Imaging device is used to obtain said useful CARS signal, carries out micro-imaging.
In the embodiment of the invention, beam split and non-linear device can be optical parametric oscillator 2 or are made up of optical splitter BS and frequency-doubling crystal 3.The conllinear focalizer mainly is made up of light combination mirror 8 and microcobjective 9.Imaging device comprises the spectrometer 22 of band pass filter 21 and band EMCCD (Electron-Multiplying CCD) detector.
Embodiment of the invention increase by one and pump light and stokes light while conllinear focus on the additional probe light of sample; This additional probe light exhausts the phonon that pump light and stokes light produce at its focal spot periphery; Form useless CARS signal; The photon that meets phase-matching condition in the detection light that postpones then with after the phonon collision at focal spot center is formed with the CARS signal of usefulness, with useless CARS Signal Separation, can obtain the spatial resolution of ultra diffraction limit.The super continuous spectrums laser that has the broadband because of employing is as pump light and stokes light, thereby obtains the complete CARS spectrum signal of molecule, realizes molecular imaging.
The above is merely preferred embodiment of the present invention, not in order to restriction the present invention, all any modifications of within spirit of the present invention and principle, being done, is equal to and replaces and improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a ultra diffraction limit coherent anti-stokes raman scattering microscopic method is characterized in that, said method comprising the steps of:
Generate ultrashort laser pulse;
Make said ultrashort laser pulse produce first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse, the wavelength of said second ultrashort laser pulse is different with the wavelength of said the 3rd ultrashort laser pulse;
Said first ultrashort laser pulse is converted into super continuous spectrums laser, second ultrashort laser pulse after postponing as surveying light, is shaped as the additional probe light that central light strength is little, ambient light is powerful with said the 3rd ultrashort laser pulse;
Make said additional probe light and said super continuous spectrums laser while conllinear focus on sample; Form useless CARS signal behind the phonon that said super continuous spectrums laser produces and the photon collision of said additional probe light; And the phonon of focal spot periphery exhausted, after colliding, the phonon of photon and focal spot central area that more said additional probe light delay arrives the detection light of sample is formed with the CARS signal of usefulness;
Obtain said useful CARS signal, carry out micro-imaging.
2. ultra diffraction limit coherent anti-stokes raman scattering microscopic method as claimed in claim 1; It is characterized in that; Said the 3rd ultrashort laser pulse is produced through nonlinear effect by said ultrashort laser pulse, and perhaps said second ultrashort laser pulse and the 3rd ultrashort laser pulse produce through nonlinear effect by said ultrashort laser pulse.
3. ultra diffraction limit coherent anti-stokes raman scattering microscopic method as claimed in claim 2; It is characterized in that said first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse produce by said ultrashort laser pulse pump light parametric oscillator;
Through the constant ultrashort laser pulse of the wavelength of said optical parametric oscillator output as said first ultrashort laser pulse; Through the ideler frequency light of said optical parametric oscillator output as said second ultrashort laser pulse, through the flashlight of said optical parametric oscillator output as said the 3rd ultrashort laser pulse.
4. ultra diffraction limit coherent anti-stokes raman scattering microscopic method as claimed in claim 1 is characterized in that, said the 3rd ultrashort laser pulse by said ultrashort laser pulse beam split after nonlinear effect produce.
5. ultra diffraction limit coherent anti-stokes raman scattering microscopic method as claimed in claim 4; It is characterized in that; Said ultrashort laser pulse is divided into three beams; As said the 3rd ultrashort laser pulse, other two bundle ultrashort laser pulses are respectively as said first ultrashort laser pulse and second ultrashort laser pulse through frequency multiplication in wherein a branch of ultrashort laser pulse.
6. like each described ultra diffraction limit coherent anti-stokes raman scattering microscopic method in the claim 1～5; It is characterized in that; Said super continuous spectrums laser comprises pump light and the stokes light that is used to produce phonon; Said pump light, stokes light and detection light intensity all are Gaussian distribution, and said additional probe light intensity distributes ringwise.
7. ultra diffraction limit coherent anti-stokes raman scattering microscopic method as claimed in claim 6 is characterized in that said super continuous spectrums laser is produced by said ultrashort laser pulse pump photon crystal optical fibre.
8. ultra diffraction limit coherent anti-stokes raman scattering microscopic method as claimed in claim 6; It is characterized in that; Each spectral power fluctuation is not more than 50% in the said super continuous spectrums laser; Each spectral component is continuous and spectral bandwidth is 10～500 nanometers, and the time range at each spectral component place is not more than 1 psec.
9. like each described ultra diffraction limit coherent anti-stokes raman scattering microscopic method in the claim 1～5, it is characterized in that more said additional probe light delay 0.5～1 psec of said detection light; The centre wavelength of the more said detection light of the centre wavelength of said additional probe light is short, and the interval of the centre wavelength of the centre wavelength of said additional probe light and said detection light is not less than the bandwidth of said useful CARS signal; The bandwidth of said detection light is not more than 5 nanometers.
10. ultra diffraction limit coherent anti-stokes raman scattering microscopic system is characterized in that said system comprises:
Femto-second laser is used to generate ultrashort laser pulse;
Beam split and non-linear device are used to make said ultrashort laser pulse to produce first ultrashort laser pulse, second ultrashort laser pulse and the 3rd ultrashort laser pulse, and the wavelength of said second ultrashort laser pulse is different with the wavelength of said the 3rd ultrashort laser pulse;
Reforming unit is used for said first ultrashort laser pulse is converted into super continuous spectrums laser;
Apparatus for shaping is used for said the 3rd ultrashort laser pulse is shaped as the additional probe light that central light strength is little, ambient light is powerful;
Deferred mount is used to regulate the time that said second ultrashort laser pulse and additional probe light arrive sample, with second ultrashort laser pulse after postponing as surveying light;
The conllinear focalizer; Be used to make said additional probe light and said super continuous spectrums laser while conllinear to focus on sample; Form useless CARS signal behind the phonon that said super continuous spectrums laser produces and the photon collision of said additional probe light; And the phonon of focal spot periphery exhausted, after colliding, the phonon of photon and focal spot central area that more said additional probe light delay arrives the detection light of sample is formed with the CARS signal of usefulness;
Imaging device is used to obtain said useful CARS signal, carries out micro-imaging.
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| CN103959045B (en) * | 2011-12-05 | 2016-06-22 | 索尼公司 | Measurement apparatus and measuring method |
| CN102540620B (en) * | 2012-02-14 | 2013-12-25 | 南昌航空大学 | Polarization interference multi-element CARS (Coherent Anti-stokes Raman Scattering) microscopic imaging method |
| CN102608100B (en) * | 2012-03-02 | 2014-04-30 | 中国科学院光电技术研究所 | System and method for CARS imaging by using four-wave mixing signals generated by optical fiber |
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