CN104698068A - High-spatial resolution laser biaxial differential confocal spectrum-mass spectrometry microimaging method and device - Google Patents
High-spatial resolution laser biaxial differential confocal spectrum-mass spectrometry microimaging method and device Download PDFInfo
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Abstract
The invention relates to a high-spatial resolution laser biaxial differential confocal spectrum-mass spectrometry microimaging method and device, and belongs to the technical fields of confocal microimaging, spectrum imaging and mass spectrometry imaging. According to the method, the biaxial differential confocal imaging technology, the mass spectrometry imaging technology and the spectral detection technology are combined, high-spatial resolution morphological imaging is performed on a sample by use of the focusing micro-spot, treated by use of a super-resolution technology, of a biaxial differential confocal system, a mass spectrometry detection system is used for performing micro-area mass spectrometry imaging on charged molecules, atoms and the like generated by performing desorption ionization on the sample with the focusing spot of a biaxial differential confocal system, a spectrum detection system is used for performing spectral imaging on plasma emission spectrum information generated by performing desorption ionization on the sample with the focusing spot of the biaxial differential confocal system, and then micro-area high-spatial resolution and high-sensitivity imaging and detection on the morphology and the components of the sample to be tested are realized by virtue of the fusion and the comparison of the detection data information. The method and the device are capable of providing a brand new technical way for high resolution imaging of the biomass spectrometry.
Description
Technical field
The invention belongs to confocal microscopic imaging technology, spectral imaging technology and mass spectrum imaging technical field, dual-axis differential confocal micro-imaging technique, mass spectrum imaging technology and spectrographic detection technology are combined, relate to a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method and device, can be used for the high-resolution imaging of biological mass spectrometry.
Technical background
Mass spectrometer (Mass Spectrometry) is ionized the component in sample, the charge atom of the different specific charges of generation, molecule or molecular fragment focused on respectively under the effect of Electric and magnetic fields and obtains the collection of illustrative plates instrument by the arrangement of mass-to-charge ratio size order.Mass spectrum imaging carries out mass spectrophotometry respectively to detect the distribution of specific mass-to-charge ratio (m/z) material to tiny area multiple in sample 2 dimensional region.
From the appearance of the substance assistant laser desorpted ionized this high sensitivity of the mid-80 in last century and high quality detection scope biological mass spectrometry imaging technique, open up field-biological mass spectrometry that mass-spectrometry one is brand-new, mass-spectrometric technique range of application is impelled to expand to the various fields of life science, the particularly application of mass spectrum in protein, nucleic acid, glycoprotein assay etc., not only for life science provides new tool, and also promote the development of mass-spectrometric technique self.
But there is following outstanding problem in existing substance assistant laser desorpted ionized mass spectrometer:
1) owing to utilizing simple Laser Focusing to carry out desorption ionization sample, thus still there is the problems such as Laser Focusing hot spot is large, mass spectrometry detection spatial resolution is not high in it;
2) cannot centering atom, molecule, intermediate ion and group etc. detect, its result constrains the accurate complete acquisition of sample mass spectrum composition;
3) mass spectrum imaging required time is long, and the relative sample of laser mass spectrometry instrument focal beam spot axial location often drifting problem occurs.
And the Obtaining Accurate of biological sample " microcell " complete component information is extremely important for life science.In fact, how detecting microcell Information in Mass Spectra is with sensitivity the important technological problems that current biological mass spectrometry field is urgently studied.
In practical work, intense pulse laser focuses on sample surfaces can make sample ionization, can inspire the atom of charged atom, molecule, molecular fragment and neutrality, molecule, intermediate ion etc.How intactly can obtain the information of the atom of charged atom, molecule, molecular fragment and neutrality, molecule, intermediate ion, the component for high accuracy analysis sample is significant.
Utilize Laser-induced Breakdown Spectroscopy (LIBS) technology can measure plasma emission spectroscopy information, obtain elemental composition in sample.Laser-induced Breakdown Spectroscopy (LIBS) technology is combined with mass spectrometry detection technology, can be used to make up in laser mass spectrometry imaging technique the deficiency that cannot obtain neutral atom, molecule and intermediate ion information.
The imaging detection mechanism of laser scanning confocal microscopy " some illumination " and " some detection ", its transverse resolution is not only made to improve 1.4 times compared with the optical microscope of equivalent parameters, but also make confocal microscope pole be convenient to combine to compress focal beam spot with super-resolution pupil filtering technique, the tight focusing technology of radial polarisation light etc., realize high-space resolution micro-imaging further.
Based on this, the present invention proposes a kind of laser dual-axis differential confocal spectrum-mass spectrum micro imaging method and device of high spatial resolution, its innovation is: will have high-space resolution ability dual-axis differential confocal microtechnic first and merge mutually with Laser-induced Breakdown Spectroscopy (LIBS) technology and mass spectrometry detection technology, and can realize imaging and the detection of sample microcell high-space resolution and highly sensitive form and component.
A kind of high-space resolution of the present invention laser dual-axis differential confocal spectrum-mass spectrum micro imaging method and device can be biological mass spectrometry high-resolution imaging and provide a brand-new effective technical way.
Summary of the invention
The object of the invention is to improve the spatial resolving power of mass spectrum imaging, suppress the drift of focal beam spot relative sample in imaging process, a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method and device are proposed, to obtaining sample component space information and function information simultaneously.The detecting function of laser dual-axis differential confocal microscope focal beam spot merges with Laser Focusing desorption ionization function by the present invention mutually, the microscopical small focal beam spot of dual-axis differential confocal through super resolution technology process is utilized to carry out the imaging of high-space resolution form to sample, utilize the charged molecule that mass spectrometry detection system produces dual-axis differential confocal microscopic system focal beam spot desorption ionization sample, atoms etc. carry out microcell mass spectrum imaging, the plasma emission spectroscopy information utilizing spectrum investigating system to produce dual-axis differential confocal microscopic system focal beam spot desorption ionization sample carries out light spectrum image-forming, and then by sample composition information that fusion and the comparison of detection data information have obtained, then imaging and the detection of sample microcell high-space resolution and highly sensitive form and component is realized.
The object of the invention is to be achieved through the following technical solutions.
A kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method of the present invention, it utilizes the focal beam spot of high-space resolution dual-axis differential confocal microscopic system to focus and imaging sample, utilize the charged molecule that mass spectrometry detection system produces dual-axis differential confocal microscopic system focal beam spot desorption ionization sample, atoms etc. carry out microcell mass spectrum imaging, the plasma emission spectroscopy utilizing spectrum investigating system to produce dual-axis differential confocal microscopic system focal beam spot desorption ionization sample detects, and then imaging and detection while then realizing sample microcell high-space resolution and highly sensitive form and component by the fusion of detection data information and compare of analysis, comprise the following steps:
Step one, make parallel beam by being shaped as annular beam after ring light generation systems, this annular beam focuses on desorption ionization on sample produce plasma plume through measuring object lens again;
Step 2, make computing machine control three-dimensional working platform drive sample along measuring surface normal direction measurement object focal point near move up and down, utilize and gather object lens, condenser lens, relaying amplifying lens and be positioned at relaying amplifying lens focal plane and carry out segmentation detection about the first light intensity point probe and the second light intensity point probe that gather the placement of optical axis symmetry to amplification Airy disk, the strength characteristics obtaining Airy disk first microcell and Airy disk second microcell is respectively first from axle confocal axial strength curve and second from the confocal axial strength curve of axle;
Step 3, subtract each other process from axle confocal axial strength curve and second from the confocal axial strength curve of axle by first and obtain dual-axis differential confocal axial strength curve;
Step 4, computing machine are according to the null position z of dual-axis differential confocal axial strength curve
avalue controls three-dimensional working platform and drives sample to move along measuring surface normal direction, makes the focal beam spot of measurement object lens focus on sample;
Step 5, utilize ionized sample suction pipe to be produced by focal beam spot desorption ionization sample plasma plume in molecule, atom and ion suck in mass spectrometry detection system and carry out mass spectrum imaging, record the Information in Mass Spectra in corresponding focal beam spot region;
Step 6, utilize by three-dimensional working platform, gather object lens, condenser lens, relaying amplifying lens and be positioned at relaying amplifying lens focal plane and about gathering the first light intensity point probe that optical axis symmetry places and the laser dual-axis differential confocal detection system that forms of the second light intensity point probe, imaging carried out to the microcell measuring object lens and focus on sample, record the shape information in corresponding focal beam spot region;
Step 7, utilize spectrum investigating system to detect the Laser-induced Breakdown Spectroscopy of collecting through splitter reflects and spectral collection lens, record the spectral information in corresponding focal beam spot region;
The Information in Mass Spectra of the Laser Focusing microcell that the spectral information of the Laser Focusing microcell that the Laser Focusing microcell shape information that laser dual-axis differential confocal detection system records by step 8, computing machine, spectrum investigating system detect simultaneously, mass spectrometry detection system record simultaneously carries out fusion treatment, then obtains the form of focal beam spot microcell, spectrum and Information in Mass Spectra;
Step 9, computing machine control three-dimensional working platform makes measurement object focal point aim at the next one region to be measured of measurand, then operates by step 2 ~ step 8, obtains the form of next focal zone to be measured, spectrum and Information in Mass Spectra;
Step 10, repetition step 9, until all tested points on sample are all measured, then utilize computing machine to carry out process and can obtain sample form, spectrum and Information in Mass Spectra.
In high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method of the present invention, comprise step one to can be and make parallel beam by being shaped as annular beam after vector beam generation systems, iris filter, this annular beam focuses on desorption ionization on sample produce plasma plume through measuring object lens again.
A kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device of the present invention, comprise pointolite, along the collimation lens that optical axis direction is placed, produce the ring light generation systems of annular beam and the focal beam spot measurement object lens to sample, comprise the collection object lens for detecting measurement object lens focal beam spot intensity of reflected light signal along gathering optical axis direction placement, condenser lens, relaying amplifying lens and be positioned at relaying amplifying lens focal plane and about gather optical axis symmetry place the first light intensity point probe and the second light intensity point probe, also comprise the dichronic mirror of exploring laser light induced breakdown spectroscopy, be positioned at the spectral collection lens of dichroic mirror light direction and be positioned at the spectrum investigating system at spectral collection lens focus place, and along measuring surface normal direction, for detecting ionized sample suction pipe and the mass spectrometry detection system of the ion body feathers component measuring object lens focal beam spot desorption ionization, incident light axis and the angle gathered between optical axis are 2 θ, and it is symmetrical about measuring surface normal.
In a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device of the present invention, comprising ring light generation systems can substitute with the vector beam generation systems of the generation vector beam placed along incident light axis direction and iris filter.
Beneficial effect
The present invention contrasts prior art, has the following advantages:
1) the dual-axis differential confocal microtechnic with high-space resolution ability is merged mutually with mass spectrometry detection technology, make the hot spot of dual-axis differential confocal micro imaging system realize focusing-detection and sample desorption ionization dual-use function, sample microcell mass spectrographic high spatial mass spectrum micro-imaging can be realized;
2) detect while Laser-induced Breakdown Spectroscopy, overcoming existing laser mass spectrometry instrument centering atom, molecule, intermediate ion and group etc. cannot carry out the deficiency that detects, can obtain microcell component information more comprehensively;
3) utilize the zero crossing of dual-axis differential confocal curve to carry out sample to focus in advance, minimum focal beam spot is made to focus on sample surfaces, sample microcell high-space resolution mass spectrometry detection and microcell micro-imaging can be realized, effectively play the potential differentiated between dual-axis differential confocal system altitude;
4) utilize dual-axis differential confocal curve zero crossing to carry out sample and focus process in advance, existing mass spectrometer can be suppressed because of the drifting problem of the relative sample of focal beam spot in long-time mass spectrum imaging;
5) utilize annular beam imaging both to have compressed the size of focal beam spot, again for mass spectrometry detection provides the best fusion of configuration aspects, the spatial resolving power of laser mass spectrometry instrument can be improved;
6) utilize the oblique incidence sounding of cross-compound arrangement light beam, overcome the defect that existing confocal microscopic imaging technology cannot suppress focal plane interference of stray light, anti-parasitic light ability is strong.
Accompanying drawing explanation
Fig. 1 is high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method of the present invention;
Fig. 2 is high-space resolution laser dual-axis differential confocal spectrum of the present invention-mass spectrum micro imaging method conversion schematic diagram;
Fig. 3 is high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method and the device schematic diagram of embodiments of the invention 1.
Wherein: 1-pointolite, 2-collimation lens, 3-parallel beam, 4-ring light generation systems, 5-ring light, 6-measures object lens, 7-incident light axis, 8-sample, 9-plasma plume, 10-gathers optical axis, 11-gathers lens, 12-condenser lens, 13-relaying amplifying lens, 14-amplifies Airy disk, 15-first light intensity point probe, 16-second light intensity point probe, 17-Airy disk first microcell, 18-Airy disk second district, 19-first is from the confocal axial strength curve of axle, 20-second is from the confocal axial strength curve of axle, 21-differential confocal axial strength curve, 22-computing machine, 23-ionized sample suction pipe, 24-mass spectrometry detection system, 25-dichronic mirror, 26-spectral collection lens, 27-spectrum investigating system, 28-three-dimensional working platform, 29-measuring surface normal, 30-CCD detector, 31-vector beam generation systems, 32-iris filter, 33-pulsed laser, 34-collector lens, 35-pin hole, 36-Optic transmission fiber, 37-outgoing beam attenuator, 38-detecting light beam attenuator.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described.
As shown in Figure 1, following examples all realize core methed of the present invention on Fig. 1 basis.
Embodiment 1
The embodiment of the present invention is based on the high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device shown in Fig. 3, this device adopts the pointolite 1 in Optic transmission fiber 36 alternate figures 1 at pulsed laser 33, collector lens 34 and collector lens 34 focus place, by the first light intensity point probe 15 and the second light intensity point probe 16 being positioned at relaying amplifying lens 13 focal plane in ccd detector 30 alternate figures 1.In the laser focusing system of Fig. 3, introduce outgoing beam attenuator 37, in laser dual-axis differential confocal detection system, introduce detecting light beam attenuator 38.
High-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device as shown in Figure 3 comprises pointolite 1, the collimation lens 2 placed along incident light axis 7 direction, outgoing beam attenuator 37, ring light generation systems 4, focal beam spot is to the measurement object lens 6 of sample 8, also comprise the collection lens 11 for detecting measurement object lens 6 focal beam spot intensity of reflected light signal along gathering the placement of optical axis 10 direction, detecting light beam attenuator 38, condenser lens 12, relaying amplifying lens 13 and the ccd detector 30 being positioned at relaying amplifying lens 13 focus, also comprise the dichronic mirror 25 for exploring laser light induced breakdown spectroscopy 39, the spectrum investigating system 27 at spectral collection lens 26 and spectral collection lens 26 focus place, and be positioned at ionized sample suction pipe 23 and the mass spectrometry detection system 24 for detecting ion body feathers 9 component measuring object lens 6 focal beam spot desorption ionization in measuring surface normal 29 direction, incident light axis 7 and the angle gathered between optical axis 10 are 2 θ, and it is symmetrical about measuring surface normal 29.
The function of main composition is as follows:
The laser focusing system be made up of to the measurement object lens 6 of sample 8 pointolite 1, the collimation lens 2 placed along incident light axis 7 direction, ring light generation systems 4, focal beam spot is for generation of the small focal beam spot exceeding diffraction limit, and this super diffraction microsize hot spot has the dual-use function measuring sample surfaces and generation surface plasma.
Precision is carried out to sample 8 focus by along gathering laser dual-axis differential confocal detection system that the collection object lens 11 in optical axis 10 direction, condenser lens 12, relaying amplifying lens 13 and the ccd detector 30 that is positioned at relaying amplifying lens 13 focus place form, and carry out imaging to measuring the microcell that object lens 6 focus on sample 8, record the shape information in corresponding focal beam spot region;
The mass spectrometry detection system be made up of ionized sample suction pipe 23 and mass spectrometry detection system 24 detects charge atom, molecule etc. in plasma plume 9 based on time-of-flight method (TOF), carry out flight time mass spectrum detection.
By the spectrum investigating system gathering object lens 11, dichronic mirror 25, the spectral collection lens 26 being positioned at dichronic mirror 25 reflected light direction and the spectrum investigating system 27 that is positioned at spectral collection lens 26 focus place and form, for detecting the Laser-induced Breakdown Spectroscopy 39 of sample 8, record the component information in corresponding focal beam spot region;
Light intensity regulating system is formed, for the spot intensity of decay focal beam spot and ccd detector 30 detection, with light intensity demand during location, accommodate sample surface by outgoing beam attenuator 37 and detecting light beam attenuator 38
The ring light transverse super-resolution system be made up of ring light generation systems 4 and measurement object lens 6, for compressing focal beam spot lateral dimension.
The tight focusing system of radial polarisation light longitudinal field be made up of vector beam generation systems 31, iris filter 32 and measurement object lens 6 is for compressing focal beam spot lateral dimension.
The three-dimensional motion system be made up of computing machine 22, three-dimensional working platform 28 can be carried out axis to sample 8 and be focused location and 3-D scanning.
The wavelength of pulsed laser 33, pulsewidth and repetition frequency can be selected as required.
The process of sample being carried out to high resolution mass spectrum imaging mainly comprises the following steps:
The light beam of step one, pulsed laser 33 outgoing collimates as parallel beam 3 after collector lens 34, Optic transmission fiber 36 and collimation lens 2, this parallel beam 3 generates annular beam 5 through outgoing beam attenuator 37, ring light generation systems 4, and annular beam 5 is focused to through measurement object lens 6 small spot exceeding diffraction limit again and is radiated on sample 8;
Step 2, utilize computing machine 22 to control three-dimensional working platform 28 to make to carry out axial scan by along gathering the laser dual-axis differential confocal detection system that the collection object lens 11 in optical axis 10 direction, condenser lens 12, relaying amplifying lens 13 and the ccd detector 30 that is positioned at relaying amplifying lens 13 focus place form to sample 8, carry out segmentation to amplification Airy disk 14 to detect, record first of corresponding Airy disk first microcell 17 and Airy disk second microcell 18 from axle confocal axial strength curve 19 and second from the confocal axial strength curve 20 of axle;
Step 3, subtract each other process from axle confocal axial strength curve 19 and second from the confocal axial strength curve 20 of axle by first and obtain dual-axis differential confocal axial strength curve 21;
Step 4, computing machine are according to the null position z of dual-axis differential confocal axial strength curve 21
avalue controls three-dimensional working platform 25 and drives sample 8 to move along measuring surface normal 29 direction, makes the focal beam spot of measurement object lens 6 focus on sample, realizes initially focusing sample 8;
Step 5, adjust irradiating light beam attenuator 37 and strengthen and measure the focal beam spot intensity of object lens 6 and make sample 8 surface produce plasma, molecule, atom and ion in the plasma plume 9 utilizing ionized sample suction pipe 23 to be produced by focal beam spot desorption ionization sample 8 suck in mass spectrometry detection system 24 and carry out mass spectrum imaging, record the Information in Mass Spectra in corresponding focal beam spot region;
Step 6, utilize by along gather optical axis 10 direction collection object lens 11, the laser dual-axis differential confocal detection system that forms of condenser lens 12, relaying amplifying lens 13 and the ccd detector 30 that is positioned at relaying amplifying lens 13 focus place carries out imaging to the microcell form of sample 8 surface plasma body feathers 9 correspondence simultaneously, record regional morphology information, detecting light beam attenuator 38 detects to avoid ccd detector 30 supersaturation for the light intensity that decays;
Step 7, simultaneously, utilizes spectrum investigating system 27 to carry out light spectrum image-forming detection to the Laser-induced Breakdown Spectroscopy 39 reflected through dichronic mirror 25 and spectral collection lens 26 are collected, records the component information in corresponding focal beam spot region;
The Information in Mass Spectra of the Laser Focusing microcell that the Laser-induced Breakdown Spectroscopy 39 of the Laser Focusing microcell that the Laser Focusing microcell shape information that laser dual-axis differential confocal detection system records by step 8, computing machine 22, spectrum investigating system 27 detect simultaneously, mass spectrometry detection system 24 detect simultaneously carries out fusion treatment, then obtains the form of focal beam spot microcell, spectrum and Information in Mass Spectra;
Step 9, computing machine 22 control three-dimensional working platform 28 makes measurement object lens 6 aim at the next one region to be measured of sample, then operates by step 2 ~ step 7, obtains form and the Information in Mass Spectra of next focal zone to be measured;
Step 10, repetition step 8, until all tested points on sample 8 are all measured, then utilize computing machine 18 to carry out data fusion and image reconstruction process, can obtain sample shape information and Information in Mass Spectra.
Embodiment 2
In high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device as shown in Figure 2, pointolite 1 can be substituted by the pin hole 35 along the pulsed laser 33 in incident light axis 7 direction, collector lens 34, collector lens 34 focus, ring light generation systems 4 can be substituted by vector beam generation systems 31, iris filter 32, and the first light intensity point probe 15 and the second light intensity point probe 16 that are positioned at relaying amplifying lens 13 focal plane are substituted by ccd detector 30.
The tight focusing system of radial polarisation light longitudinal field be made up of vector beam generation systems 31, iris filter 32 and measurement object lens 6 is for compressing focal beam spot lateral dimension.
All the other imaging measurement methods are identical with embodiment 1.
Below by reference to the accompanying drawings the specific embodiment of the present invention is described, but these explanations can not be understood to limit scope of the present invention.
Protection scope of the present invention is limited by the claims of enclosing, and any change on the claims in the present invention basis is all protection scope of the present invention.
Claims (4)
1. high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method, it is characterized in that: utilize the focal beam spot of high-space resolution dual-axis differential confocal microscopic system to focus and imaging sample, utilize the charged molecule that mass spectrometry detection system produces dual-axis differential confocal microscopic system focal beam spot desorption ionization sample, atoms etc. carry out microcell mass spectrum imaging, the plasma emission spectroscopy utilizing spectrum investigating system to produce dual-axis differential confocal microscopic system focal beam spot desorption ionization sample detects, and then imaging and detection while then realizing sample microcell high-space resolution and highly sensitive form and component by the fusion of detection data information and compare of analysis, comprise the following steps:
Step one, make parallel beam (3) by being shaped as annular beam (5) after ring light generation systems (4), this annular beam (5) focuses on the upper desorption ionization of sample (8) produce plasma plume (9) through measuring object lens (6) again;
Step 2, making computing machine (22) control three-dimensional working platform (28) drives sample (8) to move up and down along measuring surface normal (29) direction at measurement object lens (6) near focal point, utilize and gather object lens (11), condenser lens (12), relaying amplifying lens (13) and be positioned at relaying amplifying lens (13) focal plane and about gather optical axis (10) symmetry place the first light intensity point probe (15) and the second light intensity point probe (16) to amplification Airy disk (14) carry out segmentation detect, the strength characteristics obtaining Airy disk first microcell (17) and Airy disk second microcell (18) is respectively first from axle confocal axial strength curve (19) and second from the confocal axial strength curve (20) of axle,
Step 3, subtract each other process from axle confocal axial strength curve (19) and second from the confocal axial strength curve (20) of axle by first and obtain dual-axis differential confocal axial strength curve (21);
Step 4, computing machine (22) are according to the null position z of dual-axis differential confocal axial strength curve (21)
avalue controls three-dimensional working platform (28) and drives sample (8) along the motion of measuring surface normal (29) direction, makes the focal beam spot of measurement object lens (6) focus on sample (8);
Step 5, utilize ionized sample suction pipe (23) to be produced by focal beam spot desorption ionization sample (8) plasma plume (9) in molecule, atom and ion suck in mass spectrometry detection system (24) and carry out mass spectrum imaging, record the Information in Mass Spectra in corresponding focal beam spot region;
Step 6, utilize by three-dimensional working platform (28), gather object lens (11), condenser lens (12), relaying amplifying lens (13) and be positioned at relaying amplifying lens (13) focal plane and about gathering the first light intensity point probe (15) that optical axis (10) symmetry places and the laser dual-axis differential confocal detection system that forms of the second light intensity point probe (16) carries out imaging to the microcell that measurement object lens (6) focuses on sample (8), record the shape information in corresponding focal beam spot region;
Step 7, utilize spectrum investigating system (27) to detect through colour annalyzer (25) reflection and spectral collection lens (26) Laser-induced Breakdown Spectroscopy (39) of collecting, record the spectral information in corresponding focal beam spot region;
The Information in Mass Spectra of the Laser Focusing microcell that the spectral information of the Laser Focusing microcell that the Laser Focusing microcell shape information that laser dual-axis differential confocal detection system records by step 8, computing machine (22), spectrum investigating system (27) detect simultaneously, mass spectrometry detection system (24) record simultaneously carries out fusion treatment, then obtains the form of focal beam spot microcell, spectrum and Information in Mass Spectra;
Step 9, computing machine (22) control three-dimensional working platform (28) makes measurement object lens (6) focus aim at the next one region to be measured of measurand (8), then operate by step 2 ~ step 8, obtain the form of next focal zone to be measured, spectrum and Information in Mass Spectra;
Step 10, repetition step 9, until all tested points on sample (8) are all measured, then utilize computing machine (22) to carry out process and can obtain sample form, spectrum and Information in Mass Spectra.
2. a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method according to claim 1, it is characterized in that: comprise step one and can be and make parallel beam (3) by being shaped as annular beam (5) after vector beam generation systems (31), iris filter (32), this annular beam (5) focuses on the upper desorption ionization of sample (8) produce plasma plume (9) through measuring object lens (6) again.
3. high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device, it is characterized in that: comprise pointolite (1), the collimation lens (2) placed along incident light axis (7) direction, produce the ring light generation systems (4) of annular beam and the focal beam spot measurement object lens (6) to sample (8), comprise the collection object lens (11) for detecting measurement object lens (6) focal beam spot intensity of reflected light signal along gathering the placement of optical axis (10) direction, condenser lens (12), relaying amplifying lens (13) and be positioned at relaying amplifying lens (13) focal plane and about gather optical axis (10) symmetry place the first light intensity point probe (15) and the second light intensity point probe (16), also comprise the dichronic mirror (25) of exploring laser light induced breakdown spectroscopy (39), be positioned at the spectral collection lens (26) in dichronic mirror (25) reflected light direction and be positioned at the spectrum investigating system (27) at spectral collection lens (26) focus place, and along measuring surface normal (29) direction, for detecting ionized sample suction pipe (23) and the mass spectrometry detection system (24) of ion body feathers (9) component measuring object lens (6) focal beam spot desorption ionization, incident light axis (7) and the angle gathered between optical axis (10) are 2 θ, and it is symmetrical about measuring surface normal (29).
4. a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device according to claim 3, is characterized in that: comprising ring light generation systems (4) can substitute with the vector beam generation systems (31) of the generation vector beam placed along incident light axis (7) direction and iris filter (32).
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