CN1963469A - Nonlinear micro imaging method of multiphoton ionization induced by ultrashort pulse laser - Google Patents
Nonlinear micro imaging method of multiphoton ionization induced by ultrashort pulse laser Download PDFInfo
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- CN1963469A CN1963469A CN 200610129630 CN200610129630A CN1963469A CN 1963469 A CN1963469 A CN 1963469A CN 200610129630 CN200610129630 CN 200610129630 CN 200610129630 A CN200610129630 A CN 200610129630A CN 1963469 A CN1963469 A CN 1963469A
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Abstract
This invention relates to one super short impulse laser guided multiple photon non linear microscopes imaging method and its application, which uses focus super short impulse laser guidance object local surface gas or micro ionize to form micro area plasma, wherein, it adopts photoelectricity detector for plasma electromagnetic irradiation intensity within invisible light wave section; it gets object materials space change microscope images through recording object different position plasma irradiation intensity.
Description
[technical field]
The present invention relates to material analysis and micro-imaging technique field, particularly a kind of non-linear micro-imaging new method of the multiphoton ionization process based on atom, molecule and new system and application thereof.
[background technology]
In the development in science and technology in the human world, be a very important scientific research field all the time to the micro-imaging and the microcosmic material analysis of micro-object, the breakthrough in this field tends to the development of other subject and technology is produced significant impetus.Supermicroscope from the earliest optical loupes to the modern times, as scanning electron microscope, atomic force microscope and tunnel flying-spot microscope etc., appearance that we can say each new imaging technique all is the important milestone of scientific technological advance.Along with laser and appearance ultrafast, the ultra-intense laser pulse, make the further investigation of nonlinear optics and practical application become possibility.The non-linear process of light and matter interaction is applied to micro-imaging and material analysis, not only can improves the spatial resolution of optical microscopy widely and, utilize the demixing scan technology, also can carry out three-dimensional imaging the resolution of measured object component and structure.
The nonlinear optics imaging technique of having reported has multiphoton fluorescence imaging, second harmonic and third harmonic imaging, anti-Stokes microscopy, excited fluorescence launch loss microscopy etc., and wherein multiphoton fluorescence is micro-good application example all occurred with the third harmonic imaging.But, multiphoton fluorescence has only obtained using widely in biological specimen, need inject fluorescent marker under wherein a lot of situations in measured object, and the third harmonic imaging is also because its specific (special) requirements to the measured object non-linear nature has very big application limitation.
Laser-induced Breakdown Spectroscopy (LIBS) is considered to be in a kind of important new material analytical technology that a lot of aspects are better than traditional chemical material analysis method.This technology have to measured object preliminary work require low, applied widely, to characteristics such as measured object damage are little.But LIBS need use the spectroscopic analysis system of relative complex, therefore often is accompanied by problems such as whole light signal utilization factor is not high enough, amount of test data is big, the time is long.In addition, under the situation that the sensitivity of spectroscopic analysis system can not meet the demands, need to rely on the light intensity that increases irradiating laser to improve the intensity of plasma signal, this has just increased the weight of the degree of injury of measured object and scope, and serious damage can cause the reduction of spatial resolution simultaneously.So, aspect such as material analysis in micro-imaging and the micron or the microcell of sub-micrometer scale, the application of LIBS has significant limitation.
In fact, if LIBS material analysis technology and scanning confocal laser capture microdissection technology are combined, replace the spectroscopic analysis system among the LIBS and measured object carried out point by point scanning just forming a kind of new non-linear imaging technique with photodetector.
[summary of the invention]
Purpose of the present invention aims to provide a kind of non-linear micro imaging method of multiphoton ionization and dedicated system and application thereof of induced by ultrashort pulse laser.It utilizes the gas ionization of induced by ultrashort pulse laser measured object material self or near surface to produce plasma, and survey the spatial variations of measured object internal material or surface state by the radiation intensity of measuring this plasma, for a kind of brand-new technology means that provide are provided for material analysis and microscopic species.
The present invention discloses a kind of non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser for achieving the above object, it is characterized in that it may further comprise the steps:
(1) utilizes the induced by ultrashort pulse laser measured object local surfaces gas of convergence or the tiny area ionization of measured object inside, thereby form the microcell plasma;
(2) adopt the electromagnetic wave radiant intensity of plasma in visible light wave range in the photodetector ionization detection district;
(3) amplify the photosignal of exporting by photodetector with signal amplifier, and this intensity level of input Computer Storage;
(4) by controlling the relative position of measured object and ultra-short pulse laser bundle, repeat above-mentioned steps the measured object diverse location is carried out two dimensional surface scanning, or the 3-D scanning of Different Plane is surveyed, obtain the microcell plasma resonance intensity of measured object diverse location, with its input computing machine, construct the micro-image of reflection measured object material character and spatial variations thereof according to the intensity of diverse location;
(5) the grey scale signal figure to gained carries out pseudo-color coding.
The invention also discloses the system of the non-linear micro imaging method of multiphoton ionization that is exclusively used in induced by ultrashort pulse laser.Comprise ultrashort pulse laser, signal amplifier, photodetector, optical filter, dichroic mirror, catoptron, lighting source, real-time monitoring device CCD, imaging len, semi-transparent semi-reflecting lens, aperture, microcobjective, three-dimensional transfer table and computing machine is characterized in that it constitutes many light paths:
Said the 1st light path is: enter microcobjective (13) after the reflection of ultra-short pulse laser pulse through dichroic mirror (5) of ultrashort pulse laser (1) output, shine measured object (15) behind this object lens focusing;
Said the 2nd light path is: measured object (15) is propagated along the opposite direction of irradiating laser by the plasma resonance light that laser excitation produces, collect through microcobjective (13), see through dichroic mirror (5) then, behind optical filter (4), receive by highly sensitive photodetector (3), enter computing machine (17) by signal amplifier (2) again;
Said the 3rd light path is: by the illumination light that lighting source (7) sends, enter microcobjective (13) focusing and be radiated on the measured object (15) after catoptron (6) reflection; This illumination light was passed through first aperture (11) and second aperture (12) successively before importing microcobjective;
Said the 4th light path is: the illumination light of measured object (15) scattering is propagated along the opposite direction of illumination light, and is collected by microcobjective (13), passes through semi-transparent semi-reflecting lens (10) then successively and imaging len (9) enters real-time monitoring device CCD (8);
Said three-dimensional transfer table (14) is placed measured object.
By electro-motor or the three-dimensional transfer table (14) of Piezoelectric Ceramic, measured object is realized two dimension or 3-D scanning; Or the optical beam scanner (16) that is made of optical scan vibration lens is realized two dimension or the 3-D scanning of ultra-short pulse laser bundle to measured object.
The purposes of the non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser of the present invention is characterized in that the surface and the inside that are used for measured object carries out lossless detection and imaging.
The purposes of the non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser of the present invention, its feature also are to be used for composition, the structure to the measured object material, the change of properties of density is carried out nonlinear resolution.
The purposes of the non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser of the present invention, its feature are that also the detection and the pathological tissues that can be used for biological sample diagnose.
The purposes of the non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser of the present invention, its feature also are to be used for the detection of inner microdefect of optically transparent material and variations in refractive index.
The purposes of the non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser of the present invention, its feature also are and can provide important real-time detection means for the little manufacture process of novel ultra-short pulse laser.
The invention has the beneficial effects as follows: compare with LIBS, this technology has not only possessed the imaging function that LIBS does not possess, and the sensitivity that has improved detection system greatly, thereby can adopt lower excitation light intensity, reduces or even avoids damage to measured object.Simultaneously, with the nanosecond excitation source that the ultrashort laser pulse of femtosecond or subpicosecond replaces LIBS to adopt usually, the energy of pulse has reduced widely when this means same excitation light intensity, thereby has further reduced the damage to measured object.Comparing with scanning confocal laser capture microdissection technology, is the non-linear detection imaging of a kind of height on this technological essence, therefore be spatial resolution, or the resolution of material behavior on all can have clearly and improve.For the situation of the space microscopic appearance of not only being concerned about measured object but also care measured object changes in material properties, the ultra-short pulse laser nonlinear plasma microtechnic that we propose will have very big advantage.This technology can be used for the surface of various materials such as metal, semiconductor, stupalith and detects, also can be used for optical material surface and the character of inside and the detection of structure, more importantly, this technology also will be significant for the detection and the microscopic diagnosis of biosome pathology.
Method and system of the present invention is applicable to the detection of various material shapes such as solid, liquid, gas and composition.The present invention has higher sensitivity to measured object material composition and structural change, and spatial resolution can reach micron or sub-micrometer scale.For the uniform measured object of material character surface, the present invention can provide the surface profile image of sub-micrometer scale.In the context of detection of the inside of optically transparent material microdefect, the present invention will have important use.In addition, use the sex change of induced with laser optics material at present, produce variations in refractive index, (light) manufacturing technology of optoelectronic devices such as preparation three-dimensional optical waveguide, fiber grating is fast-developing.The present invention can provide important real-time detection means for the little manufacture process of this novel laser, produces critical impetus thereby laser is prepared various micro photo electric devices.
Because the present invention possesses nonlinear material analysis and resolution characteristic, can reduce to greatest extent simultaneously in the detection process the degree of injury of measured object, its context of detection at biological specimen microexamination and pathological tissues has special advantages.Biological tissue and biological cell are transparent mostly, and the formation of various institutional frameworks is more approaching again, so traditional optical imaging method is difficult to distinguish, common dyeing or the fluorescent method that adopts, make imaging process become complicated on the one hand, also can cause artificial bio-toxicity in some cases on the other hand.Because the present invention has high-sensitive changes in material resolution characteristic, can attempt under achromophil situation biological specimen being carried out microexamination and analysis, therefore, this will be a kind of very promising biology and medical research instrument.
Because ionization process belongs to the high-order nonlinear process, thereby can carry out nonlinear resolution to change of properties such as the composition of measured object material, structure, density.The measured object surface is being carried out in the imaging process, because the plasma signal is near the ionisation of gas of inducing that comes from the measured object surface, and ionization does not take place in measured object itself, so can realize lossless detection and imaging.
[description of drawings]
Fig. 1 is a system schematic of the present invention;
Fig. 2 is the nitrogen gas plasma radiation spectrum of induced by ultrashort pulse laser;
Fig. 3 is the variation relation of plasma signal with excitation laser intensity;
Fig. 4 is the cross-sectional view of standard single-mode fiber;
Fig. 5 is that ultra-short pulse laser is at the inner microstructure image of inscribing of optical glass;
Fig. 6 is the microstructure image that ultra-short pulse laser is inscribed in glass sheet surface;
Fig. 7 is the micro-image of human body oral mucosa epithelial cell.
Wherein: 1. ultrashort pulse laser; 2. signal amplifier; 3. photodetector; 4. optical filter; 5. dichroic mirror; 6. catoptron; 7. lighting source; 8. real-time monitoring device (CCD); 9. imaging len; 10. semi-transparent semi-reflecting lens; 11. first aperture; 12. second aperture; 13. microcobjective; 14. three-dimensional transfer table; 15. measured object; 16. optical beam scanner; 17. computing machine.
Be described in detail with reference to accompanying drawing below in conjunction with embodiments of the invention.
[embodiment]
The non-linear micro imaging method of the multiphoton ionization of induced by ultrashort pulse laser of the present invention may further comprise the steps:
(1) utilizes the induced by ultrashort pulse laser measured object local surfaces gas of convergence or the tiny area ionization of measured object inside, thereby form the microcell plasma.Wherein: ultra-short pulse laser be pulsewidth be between 10fs and the 1ps, single pulse energy between 0.1nJ and the 10 μ J, wavelength coverage is in the near infrared ultrashort pulse between the 700nm to 2500nm: after ultra-short pulse laser is focused on by the microcobjective of a numerical aperture (N.A.) 0.1-1.5, excite the measured object self-ionization or induce air ionization, measured object or the air ionization radiation plasma light of measured object near surface.
(2) adopt the electromagnetic wave radiant intensity of plasma in visible light wave range in the photodetector ionization detection district.Be that measured object or air ionization radiation plasma light are collected through the microcobjective of numerical aperture 0.1-1.5, after the process optical filter filters residual excitation laser again, survey by photodetector.
(3) amplify the photosignal of exporting by photodetector with signal amplifier, and this intensity level of input Computer Storage.
(4) by controlling the relative position of measured object and ultra-short pulse laser bundle, repeat above-mentioned steps the measured object diverse location is carried out scanning probe, obtain the microcell plasma resonance intensity of measured object diverse location, with its input computing machine, construct the micro-image of reflection measured object material character and spatial variations thereof according to the intensity of diverse location.
In addition, before or after the measured object scanning, can introduce illumination light irradiation measured object by inserting spectroscope or catoptron, the illumination light of measured object scattering by the semi-transparent semi-reflecting mirror reflection of inserting after through imaging len imaging on real-time monitoring device CCD, in monitor, monitor.
Imaging process can be carried out in atmosphere, also measured object can be placed in the inert gas filled closed container and carry out.
System's (as shown in Figure 1) of the non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser, by ultrashort pulse laser, signal amplifier, photodetector, optical filter, dichroic mirror, catoptron, lighting source, real-time monitoring device CCD, imaging len, semi-transparent semi-reflecting lens, aperture, microcobjective, compositions such as three-dimensional transfer table and computing machine, and constitute four light paths:
Said the 1st light path is: enter microcobjective (13) after the reflection of ultra-short pulse laser pulse through dichroic mirror (5) of ultrashort pulse laser (1) output, shine measured object (15) behind this object lens focusing;
Said the 2nd light path is: measured object (15) is propagated along the opposite direction of irradiating laser by the plasma resonance light that laser excitation produces, collect through microcobjective (13), see through dichroic mirror (5) then, behind optical filter (4), receive by highly sensitive photodetector (3), enter computing machine (17) by signal amplifier (2) again.Wherein said photodetector photomultiplier, photodiode, avalanche-type photodiode, CCD and enhancement mode CCD etc.; Said signal amplifier can be lock-in amplifier or door integrator, current amplifier etc.
Said the 3rd light path is: by the illumination light that lighting source (7) sends, enter microcobjective (13) focusing and be radiated on the measured object (15) after catoptron (6) reflection; This illumination light was passed through first aperture (11) and second aperture (12) successively before importing microcobjective, with femtosecond light furnishing conllinear.Said illumination light is the visible light near collimation, especially can be that the wavelength that process suitably collimates is the He-Ne Lasers of 632.8nm; But but both also uneven Jiao of furnishing or close on parfocalization of furnishing and ultra-short pulse laser parfocalization of illumination light.
Said the 4th light path is: the illumination light of measured object (15) scattering is propagated along the opposite direction of illumination light, and is collected by microcobjective (13), passes through semi-transparent semi-reflecting lens (10) then successively and imaging len (9) enters real-time monitoring device CCD (8);
Said three-dimensional transfer table (14) is used for placing measured object.Three-dimensional transfer table (14) is the three-dimensional transfer table by electro-motor or Piezoelectric Ceramic, and measured object is realized two dimension or 3-D scanning; Or the optical beam scanner (16) that is made of optical scan vibration lens is realized two dimension or the 3-D scanning of ultra-short pulse laser bundle to measured object.
The present invention utilizes the gas of induced by ultrashort pulse laser measured object self or near surface to produce the plasma of local, and the radiation intensity that adopts photodetector to survey this plasma detects near the measured object material character the focus.By writing down the plasma resonance intensity level of different measured objects position, provide the analysis of digital microscopy images of differentiating the material character spatial variations.
Ultrashort pulse laser is as excitation source, and its exportable centre wavelength is the laser pulse of 2mJ, minimum pulse width 50fs at 800nm, maximum single pulse energy.This laser is attenuated back (be attenuated to usually single pulse energy for receive burnt magnitude) through entering the microcobjective of high-NA behind dichroic mirror, through this object lens focusing in the air of transparent measured object inside or measured object near surface.In the focus place and the adjacent domain thereof of laser, measured object (or air) can and produce plasma, the compound radiation that causes visible light wave range of electronics and ion in this plasma by laser excitation ionization.Wherein, a part of radiant light is propagated along the opposite direction of irradiating laser, and is collected by microcobjective, sees through semi-transparent semi-reflecting lens then, and the process optical filter is received by highly sensitive photodetector after filtering remaining ultra-short pulse laser.
Photodetector is sent into signal amplifier after changing the light signal that receives into the weak current signal, sends into the data collecting card in the computing machine after signal is exaggerated, and signal intensity of computer recording also stores it.The computer control scanister moved measured object or controls relatively moving of ultra-short pulse laser bundle by optical beam scanner after this process was finished, and repeated aforesaid operations in new measured object position, obtained the plasma signal intensity of this position.By measured object diverse location place is carried out scanning probe, obtain the plasma intensity at corresponding diverse location place like this, finally constitute the micro-gray level image or the pseudo color image of a width of cloth measured object.In order to monitor imaging process in real time, before or after the plasma imaging, can introduce illumination light by inserting catoptron, and insert semi-transparent semi-reflecting lens, measured object scattered light process imaging len is imaged on the CCD, monitor by monitor.
Move forward and backward measured object along the laser propagation direction, can regulate the relative distance between laser spot and the measured object surface.If laser spot is adjusted to transparent measured object inside, plasma is from the ionization of measured object material self at this moment, by the transversal scanning measured object and write down the pairing plasma signal intensity of diverse location, just can obtain differentiating the analysis of digital microscopy images of measured object internal material spatial variations, can differentiate the variation of material composition like this, also can differentiate the variation of character such as material structure and density.If laser spot is adjusted to beyond the measured object surface, allow the focus and the measured object surface of laser that certain distance is arranged, this moment induced with laser photoelectron at the effect meeting of laser air ionization with near surface, by the transversal scanning measured object and write down the pairing air plasma signal intensity of diverse location and obtain analysis of digital microscopy images, can differentiate the spatial variations of measured object surfacing character or pattern.
To the measured object surface imaging time, the influence between laser spot and the surface apart from Stimulated Light parameter (comprising intensity, focused condition etc.) and measured object character (mainly being damage threshold), its span generally at 2 μ m between the 30 μ m.If distance is too big, measured object can be too little to the air ionization influence, if distance is too little, laser can break measured object again.
Adopt among the present invention tens to the ultrashort laser pulse of hundreds of femtosecond as exciting light, the plasma resonance that it excited is the nitrogen plasma spectrum of induced by ultrashort pulse laser based on continuous spectrum as shown in Figure 2.
Because the main mechanism of induced by ultrashort pulse laser measured object ionization is multiphoton ionization, it is a kind of high-order nonlinear process, the present invention is actually a kind of optical material nonlinear analysis and micro imaging method like this, and plasma signal as shown in Figure 3 is with the variation relation of excitation laser intensity.Pure SiO
2The standard fiber covering that constitutes and by the SiO that mixes germanium (Ge)
2The fibre core that constitutes produces the plasma signal and excitation light intensity is high order power relation, and, corresponding same excitation light intensity, isoionic signal difference is very big, and this explanation plasma signal is very responsive to the variation of material.Simultaneously, this high-order nonlinear process makes the resolution that obtains inferior diffraction limit become possibility, add that the focal spot itself after the high power microcobjective focuses on is very little, this technology can reach the resolution of micron even sub-micrometer scale, and can carry out the computed tomography scanning of different depth, constitute three-dimension space image.
Accompanying drawing 4 (a) is differentiated the example that measured object material composition changes, pure SiO for the present invention
2The standard fiber covering that constitutes and by the SiO that mixes germanium (Ge)
2The fibre core that constitutes has been distinguished significantly.Used systematic parameter is: laser single-pulse energy 16nJ, and 10 * microcobjective of N.A.=0.25 focuses on, and sweep spacing is 3 μ m.Accompanying drawing 4 (b) amplifies the displaing micro picture that 500 * back is taken by digital camera for adopting the OlympusBX51 microscope.The optics presentation that adopts conventional microscope can only distinguish this changes in material is a change of refractive, because the refractive index difference of fibre cladding and fibre core is little, so the contrast of displaing micro picture is very poor.This illustrates the more directly composition variation of detecting material inside of this non-linear detection means of the present invention, and its contrast ability will be much better than conventional optical microscope.
When laser during in transparent measured object internal focus, the atom in the measured object material (or molecule) will be ionized and form in the part plasma.By mobile measured object, and the pairing plasma signal intensity of record different spatial, will obtain differentiating the micro-image of material character spatial variations.As accompanying drawing 5 (a), the microstructure of being inscribed in glass by ultra-short pulse laser can clearly be shown.As a same reason, traditional optical microscope is far short of what is expected to the separating capacity of the material sex change of this small variations in refractive index, shown in accompanying drawing 5 (b).The experiment parameter that obtains Fig. 5 (a) is: laser single-pulse energy is 16nJ, and focus lamp is 10 * microcobjective of N.A.=0.25, and sweep spacing is 3 μ m.Fig. 5 (b) is that Olympus BX51 microscope amplifies 250 * back by the digital camera picture shot.
Accompanying drawing 6 is at the micro-image of glass surface by a microstructure of ultra-short pulse laser ablation technology preparation.Wherein Fig. 6 (a) is the image that present technique obtains, and Fig. 6 (b) is the image that the conventional microscope picture obtains.Relatively two figure as can be seen, figure (a) has tangible depth resolution and contrast ability, this point is more obvious the left-half reflection of image.The parameter that obtains the image of Fig. 6 (a) is: laser single-pulse energy is 300nJ, and focus lamp is 40 * microcobjective of N.A.=0.65, and laser spot is 16 μ m apart from glass surface, and sweep spacing is 3 μ m.Fig. 6 (b) be Olympus BX51 microscope amplify 250 * by the digital camera picture shot.The physical size of Fig. 6 (a) and (b) correspondence is 150 * 150 μ m
2
Accompanying drawing 7 is depicted as the epithelial micro-image of human oral cavity, wherein the cell interior structure that obtains when cell interior for laser spot of Fig. 7 (a); Fig. 7 (b) is near the cell surface condition diagram picture that focus obtains by air plasma cell surface, because the variation of cell surface material character is very little, Fig. 7 (b) reality has reflected the surface topography of cell; The parameter that obtains the image of Fig. 7 (a) and 7 (b) is: laser single-pulse energy is 12nJ, and focus lamp is 40 * microcobjective of N.A.=0.65, and sweep spacing is 1 μ m.(c) and (d) be respectively (a) and pseudo color image (b), the physical size of (a) and (b), (c), (d) correspondence is 100 * 100 μ m
2
Claims (17)
1, the non-linear micro imaging method of a kind of multiphoton ionization of induced by ultrashort pulse laser is characterized in that it may further comprise the steps:
(1) utilizes the induced by ultrashort pulse laser measured object local surfaces gas of convergence or the tiny area ionization of measured object inside, thereby form the microcell plasma;
(2) adopt the electromagnetic wave radiant intensity of plasma in visible light wave range in the photodetector ionization detection district;
(3) amplify the photosignal of exporting by photodetector with signal amplifier, and this intensity level of input Computer Storage;
(4) by controlling the relative position of measured object and ultra-short pulse laser bundle, repeat above-mentioned steps and measured object is carried out two dimensional surface scanning, or the 3-D scanning of Different Plane is surveyed, obtain the microcell plasma resonance intensity of measured object diverse location, with its input computing machine, construct the micro-image of reflection measured object material character and spatial variations thereof according to the intensity of diverse location.
(5) the grey scale signal figure to gained carries out pseudo-color coding.
2, the non-linear micro imaging method of the multiphoton ionization of induced by ultrashort pulse laser according to claim 1, it is characterized in that in the said step (1), ultra-short pulse laser be pulsewidth between 10fs and the 1ps, single pulse energy between 0.1nJ and 10 μ J, wavelength coverage is in the near infrared ultrashort pulse between the 700nm to 2500nm.
3, the non-linear micro imaging method of the multiphoton ionization of induced by ultrashort pulse laser according to claim 1 and 2, it is characterized in that in the said step (1), ultra-short pulse laser is after the interior microcobjective of 0.1 and 1.5 scopes focuses on by a numerical aperture (N.A.), excite the measured object self-ionization or induce the air ionization of measured object near surface, measured object or air ionization produce plasma resonance.
4, the non-linear micro imaging method of the multiphoton ionization of induced by ultrashort pulse laser according to claim 3 is characterized in that to the measured object surface imaging time, the distance between said laser spot and the measured object surface at 2 μ m between the 30 μ m.
5, the non-linear micro imaging method of the multiphoton ionization of induced by ultrashort pulse laser according to claim 1 and 2, it is characterized in that in the said step (2), the plasma light of measured object or the radiation of air ionization institute is that the interior microcobjective of 0.1 and 1.5 scopes is collected through numerical aperture (N.A.), after filtering residual excitation laser through optical filter again, survey by photodetector.
6, the non-linear micro imaging method of the multiphoton ionization of induced by ultrashort pulse laser according to claim 1 and 2, it is characterized in that before said measured object is scanned, introduce illumination light irradiation measured object by inserting spectroscope or catoptron, the illumination light of measured object scattering by the semi-transparent semi-reflecting mirror reflection of inserting after through imaging len imaging on real-time monitoring device CCD, in monitor, monitor.
7, the non-linear micro imaging method of the multiphoton ionization of induced by ultrashort pulse laser according to claim 1 is characterized in that: imaging process can be carried out in atmosphere, also measured object can be placed in the inert gas filled closed container and carry out.
8, a kind of system of the non-linear micro imaging method of multiphoton ionization of the induced by ultrashort pulse laser that is exclusively used in claim 1 comprises ultrashort pulse laser, signal amplifier, photodetector, optical filter, dichroic mirror, catoptron, lighting source, real-time monitoring device CCD, imaging len, semi-transparent semi-reflecting lens, aperture, microcobjective, three-dimensional transfer table and computing machine is characterized in that:
Said the 1st light path is: enter microcobjective (13) after the reflection of ultra-short pulse laser pulse through dichroic mirror (5) of ultrashort pulse laser (1) output, shine measured object (15) behind this object lens focusing;
Said the 2nd light path is: measured object (15) is propagated along the opposite direction of irradiating laser by the plasma resonance light that laser excitation produces, collect through microcobjective (13), see through dichroic mirror (5) then, behind optical filter (4), receive by highly sensitive photodetector (3), enter computing machine (17) by signal amplifier (2) again;
Said the 3rd light path is: by the illumination light that lighting source (7) sends, enter microcobjective (13) focusing and be radiated on the measured object (15) after catoptron (6) reflection; This illumination light was passed through first aperture (11) and second aperture (12) successively before importing microcobjective;
Said the 4th light path is: the illumination light of measured object (15) scattering is propagated along the opposite direction of illumination light, and is collected by microcobjective (13), passes through semi-transparent semi-reflecting lens (10) then successively and imaging len (9) enters real-time monitoring device CCD (8);
Said three-dimensional transfer table (14) is placed measured object.
9, the system of the non-linear micro imaging method of multiphoton ionization of induced by ultrashort pulse laser according to claim 8 is characterized in that by electro-motor or the three-dimensional transfer table (14) of Piezoelectric Ceramic measured object being realized two dimension or 3-D scanning; Or the optical beam scanner (16) that is made of optical scan vibration lens is realized two dimension or the 3-D scanning of ultra-short pulse laser bundle to measured object.
10, according to Claim 8 or the system of the non-linear micro imaging method of multiphoton ionization of 9 described induced by ultrashort pulse laser, it is characterized in that said photodetector can be photomultiplier, photodiode, avalanche-type photodiode, CCD and enhancement mode CCD.
11, according to Claim 8 or the system of the non-linear micro imaging method of multiphoton ionization of 9 described induced by ultrashort pulse laser, it is characterized in that said signal amplifier can be lock-in amplifier or door integrator, current amplifier.
12, according to Claim 8 or the system of the non-linear micro imaging method of multiphoton ionization of 9 described induced by ultrashort pulse laser, it is characterized in that said illumination light is the visible light near collimation, especially can be that the wavelength of the suitable collimation of process is the He-Ne Lasers of 632.8nm; But but both also uneven Jiao of furnishing or close on parfocalization of furnishing and ultra-short pulse laser parfocalization of illumination light.
13, the purposes of the non-linear micro imaging method of multiphoton ionization of the induced by ultrashort pulse laser of claim 1 is characterized in that the surface and the inside that can be used for measured object carries out lossless detection and imaging.
14, the purposes of the non-linear micro imaging method of multiphoton ionization of the induced by ultrashort pulse laser of claim 1 is characterized in that being used for composition, the structure to the measured object material, the change of properties of density is carried out nonlinear resolution.
15, the purposes of the non-linear micro imaging method of multiphoton ionization of the induced by ultrashort pulse laser of claim 1 is characterized in that the detection and the pathological tissues that can be used for biological sample diagnose.
16, the purposes of the non-linear micro imaging method of multiphoton ionization of the induced by ultrashort pulse laser of claim 1 is characterized in that being used for the detection of inner microdefect of optically transparent material and variations in refractive index.
17, the purposes of the non-linear micro imaging method of multiphoton ionization of the induced by ultrashort pulse laser of claim 1 is characterized in that and can provide important real-time detection means for the little manufacture process of novel ultra-short pulse laser.
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