CN104698068B - 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, by dual-axis differential
Confocal microscopic imaging technology, mass spectrum imaging technology and spectrographic detection technology combine, and are related to a kind of high-space resolution laser twin shaft
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 spectrograph (Mass Spectrometry) is to ionize the component in sample, makes the different charge-mass ratios of generation
Charge atom, molecule or molecular fragment focus on respectively in the presence of electric field and magnetic field and obtain being arranged by mass-to-charge ratio size order
The collection of illustrative plates instrument of row.Mass spectrum imaging is that to carry out mass spectral analyses respectively to multiple tiny areas in sample 2 dimensional region specific to detect
The distribution of mass-to-charge ratio (m/z) material.
Give birth to from the substance assistant laser desorpted ionized this high sensitivity of last century the mid-80 and high quality detection scope
The appearance of material spectral imaging technology, has opened up the brand-new field-biological mass spectrometry of one, mass spectrometry, promotes mass-spectrometric technique application model
Enclose the various fields for expanding to life science, particularly mass spectrum answering at aspects such as protein, nucleic acid, glycoprotein assays
With, not only new tool is provided for life science, and also promote the development of mass-spectrometric technique itself.
But existing substance assistant laser desorpted ionized mass spectrograph there are problems that following prominent：
1) because using simple laser focusing, come desorption ionization sample, thus it is still suffered from, and laser focusing hot spot is big, matter
The problems such as spectrum space exploration resolving power is not high；
2) cannot centering atom, molecule, intermediate ion and group etc. detected, its result constrains sample mass spectrum composition
Accurate complete acquisition；
3) long the time required to mass spectrum imaging, laser mass spectrometry instrument focal beam spot axial location often drifts about with respect to sample
Problem.
And the accurate acquisition of the complete component information of biological sample " microcell " is extremely important for life science has
Meaning.In fact, how to detect the important technology that microcell Information in Mass Spectra is that current biological mass spectrometry field is urgently studied with sensitivity
Problem.
In practical work, intense pulse laser focuses on sample surfaces and can make sample ionization, can inspire powered atom, divide
Atom, molecule, intermediate ion of son, molecular fragment and neutrality etc..How powered atom, molecule, molecule are intactly obtained
Atom, molecule, the information of intermediate ion of fragment and neutrality, it is significant for the component of high accuracy analysis sample.
Using the measurable plasma emission spectroscopy information of LIBS (LIBS) technology, unit in sample is obtained
Plain composition.By LIBS (LIBS) technology in combination with mass spectrometry detection technology, can be used to make up laser mass spectrometry into
As the deficiency that cannot obtain neutral atom, molecule and intermediate ion information in technology.
Laser scanning confocal microscopy " point illumination " and the imaging detection mechanism of " point detection ", not only make its transverse resolution more same
Isoparametric optical microscope improves 1.4 times, but also confocal microscope pole is easy to and super-resolution pupil filtering technique, radial direction
Polarized light tightly focused technology etc. combines to compress focal beam spot, further realizes high-space resolution micro-imaging.
Based on this, the present invention proposes a kind of laser dual-axis differential confocal spectrum-mass spectrum micro-imaging of high spatial resolution
Method and apparatus, its innovation is：First will be with high-space resolution ability dual-axis differential confocal microtechnique and induced with laser
Breakdown spectral (LIBS) technology and mass spectrometry detection technology are blended, and are capable of achieving sample microcell high-space resolution and highly sensitive shape
The imaging of state and component and detection.
A kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method of the present invention can make a living with device
Material spectrum high-resolution imaging provides a brand-new effective technical way.
The content of the invention
The purpose of the present invention be improve mass spectrum imaging spatial resolving power, suppress imaging process in focal beam spot with respect to sample
The drift of product, proposes a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method and device, to same
When obtain sample component space information and function information.The present invention is by laser dual-axis differential confocal microscope focal beam spot
Detecting function is blended with laser focusing desorption ionization function, using the dual-axis differential confocal microscope of Jing super resolution technologies process
Small focal beam spot high-space resolution form imaging is carried out to sample, it is micro- to dual-axis differential confocal using mass spectrometry detection system
System focal beam spot desorption ionization sample and charged molecule, atom for producing etc. carry out microcell mass spectrum imaging, using spectrographic detection
The plasma emission spectroscopy information that system is produced to dual-axis differential confocal microscopic system focal beam spot desorption ionization sample is entered
Row light spectrum image-forming, then passes through again the fusion of detection data information and obtains the sample composition information for completing with comparing, and realizes then
The imaging of sample microcell high-space resolution and highly sensitive form and component and detection.
The purpose of the present invention is 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 is using high
The focal beam spot of spatial discrimination dual-axis differential confocal microscopic system is focused and imaging to sample, using mass spectrometry detection system pair
Dual-axis differential confocal microscopic system focal beam spot desorption ionization sample and charged molecule, atom for producing etc. carry out microcell mass spectrum into
Picture, dual-axis differential confocal microscopic system focal beam spot desorption ionization sample is produced plasma using spectrum investigating system
Emission spectrum is detected, and the fusion that detection data information is then passed through again realizes that sample microcell is high then with analysis is compared
Spatial discrimination and highly sensitive form and imaging while component and detection, comprise the following steps：
Step one, collimated light beam is set to be shaped as annular beam after ring light generation system, again Jing is surveyed the annular beam
Amount object lens focus on desorption ionization on sample and produce plasma plume；
Step 2, computer controls three-dimensional working platform is set to drive sample burnt in measurement object lens along measuring surface normal direction
Point is nearby moved up and down, using collection object lens, condenser lenses, relaying amplifying lens and positioned at relaying amplifying lens focal plane and with regard to
The first symmetrically placed light intensity point probe of collection optical axis and the second light intensity point probe carry out segmentation detection to amplifying Airy disk,
The strength characteristics for obtaining the microcell of Airy disk first and the microcell of Airy disk second are respectively the first off-axis confocal axial strength song
Line and the second off-axis confocal axial strength curve；
Step 3, the first off-axis confocal axial strength curve and the second off-axis confocal axial strength curve are subtracted each other process
To dual-axis differential confocal axial strength curve；
The dead-center position z of step 4, computer foundation dual-axis differential confocal axial strength curveAValue control three-dimensional working platform
Sample is driven to move along measuring surface normal direction, the focal beam spot for making measurement object lens is focused on sample；
In step 5, the plasma plume produced focal beam spot desorption ionization sample using ionized sample suction pipe
Mass spectrum imaging is carried out in molecule, atom and ion suction mass spectrometry detection system, the Information in Mass Spectra in correspondence focal beam spot region is measured；
Step 6, using by three-dimensional working platform, collection object lens, condenser lenses, relaying amplifying lens and positioned at relaying amplify
Lens focal plane and the laser with regard to gathering the first symmetrically placed light intensity point probe of optical axis and the second light intensity point probe composition
Dual-axis differential confocal detection system focuses on the microcell of sample to measuring object lens and is imaged, and measures correspondence focal beam spot area
The shape information in domain；
Step 7, the reflection of Jing colour annalyzers and spectral collection lens are collected laser-induced breakdown using spectrum investigating system
Spectroscopically detectable, measures the spectral information in correspondence focal beam spot region；
Laser focusing microcell shape information, light that step 8, computer measure laser dual-axis differential confocal detection system
The laser focusing microcell that the spectral information of the laser focusing microcell that spectrum detection system is detected simultaneously, mass spectrometry detection system are measured simultaneously
Information in Mass Spectra carry out fusion treatment, form, spectrum and the Information in Mass Spectra of focal beam spot microcell are obtained then；
Step 9, computer controls three-dimensional working platform make measurement object focal point be directed at the area next to be measured of sample
Domain, is then operated by step 2~step 8, obtains form, spectrum and the Information in Mass Spectra of next focal zone to be measured；
The all tested points of step 10, repeat step nine on sample are measured, and are then entered using computer
Row processes and sample form, spectrum and Information in Mass Spectra is obtained.
In high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method of the present invention, including step one can
To make collimated light beam occur to be shaped as annular beam after system, iris filter by vector beam, again Jing is surveyed the annular beam
Amount object lens focus on desorption ionization on sample and produce plasma plume.
A kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device of the present invention, including a light
There is system and focal beam spot to sample in source, the collimating lens along optical axis direction placement, the ring light of generation annular beam
Measurement object lens, including adopting for detection measurement object lens focal beam spot intensity of reflected light signal along what collection optical axis direction was placed
Collection object lens, condenser lenses, relaying amplifying lens and positioned at relaying amplifying lens focal plane and symmetrically placed with regard to gathering optical axis the
One light intensity point probe and the second light intensity point probe, dichroic mirror also including exploring laser light induced breakdown spectroscopy, positioned at color separation
The spectral collection lens and the spectrum investigating system at spectral collection lens focus of mirror reflection light direction, and along measuring surface
Normal direction, the ionized sample suction pipe for detecting the ion body feathers component for measuring object lens focal beam spot desorption ionization and mass spectrum are visited
Examining system, the angle between incident light axis and collection optical axis is 2 θ, and symmetrical with regard to 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, including ring
There is system and system and pupil filter can occur with the vector beam of the generation vector beam placed along incident light axis direction in shape light
Ripple device is substituted.
Beneficial effect
Present invention contrast prior art, with advantages below：
1) the dual-axis differential confocal microtechnique with high-space resolution ability and mass spectrometry detection technology are blended, is made double
The hot spot of axle differential confocal microscopic imaging system realizes focusing-detection and sample desorption ionization dual-use function, is capable of achieving sample microcell
Mass spectrographic high spatial mass spectrum micro-imaging；
2) while LIBS detect, overcome existing laser mass spectrometry instrument cannot centering atom, molecule,
The deficiency that intermediate ion and group etc. are detected, it is possible to obtain more comprehensively microcell component information；
3) carry out sample using the zero crossing of dual-axis differential confocal curve to focus in advance, make minimum focal beam spot focus on sample
Product surface, is capable of achieving sample microcell high-space resolution mass spectrometry detection and microcell micro-imaging, effectively plays dual-axis differential confocal
The potential differentiated between system altitude；
4) carry out sample using dual-axis differential confocal curve zero crossing and focus process in advance, existing mass spectrograph can be suppressed because of length
Drifting problem of the focal beam spot with respect to sample in time mass spectrum imaging；
5) size of focal beam spot had both been have compressed using annular beam imaging, and structure side had been provided for mass spectrometry detection
The optimal fusion in face, can improve the spatial resolving power of laser mass spectrometry instrument；
6) using cross-compound arrangement light beam oblique incidence sounding, overcoming existing confocal microscopic imaging technology cannot suppress focal plane miscellaneous
The defect of astigmatism interference, resists spuious light ability strong.
Description of the drawings
Fig. 1 is the high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method of the present invention；
Fig. 2 is that the high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method conversion of the present invention is illustrated
Figure；
Fig. 3 is the high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method of embodiments of the invention 1
With schematic device.
Wherein：There is system, 5- ring lights, 6- measurements in 1- point sources, 2- collimating lens, 3- collimated light beams, 4- ring lights
Object lens, 7- incident light axis, 8- samples, 9- plasma plumes, 10- collection optical axis, 11- collection lens, 12- condenser lenses,
13- relaying amplifying lenses, 14- amplify Airy disk, 15- the first light intensity point probes, 16- the second light intensity point probes, 17- Airies
The microcell of speckle first, the area of 18- Airy disks second, the off-axis confocal axial strength curves of 19- first, the off-axis confocal axial directions of 20- second are strong
Write music line, 21- differential confocal axial strength curves, 22- computers, 23- ionized sample suction pipes, 24- mass spectrometry detection systems, 25-
Dichroic mirror, 26- spectral collection lens, 27- spectrum investigating systems, 28- three-dimensional working platforms, 29- measuring surface normals, 30-CCD detections
There is system, 32- iris filters, 33- pulse lasers, 34- collecting lenses, 35- pin holes, 36- and pass in device, 31- vector beams
Light optical fiber, 37- outgoing beam attenuators, 38- detection beam attenuators.
Specific embodiment
With reference to the accompanying drawings and examples the invention will be further described.
The core methed of the present invention is as shown in figure 1, following examples are realized on the basis of Fig. 1.
Embodiment 1
The embodiment of the present invention is based on the high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro-imaging shown in Fig. 3
Device, the device is substituted in Fig. 1 using the Optic transmission fiber 36 of pulse laser 33, collecting lenses 34 and the focal point of collecting lenses 34
Point source 1, by ccd detector 30 substitute in Fig. 1 positioned at relaying the focal plane of amplifying lens 13 the first light intensity point probe 15
With the second light intensity point probe 16.Outgoing beam attenuator 37 is introduced in the laser focusing system of Fig. 3, in laser dual-axis differential
Detection beam attenuator 38 is introduced in confocal detection system.
High-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device as shown in Figure 3 includes point source
1, there is system 4, focal beam spot and arrive in the collimating lens 2, outgoing beam attenuator 37, ring light along the placement of the direction of incident light axis 7
The measurement object lens 6 of sample 8, also include anti-for the detection measurement focal beam spot of object lens 6 along the placement of collection optical axis 10 direction
The collection lens 11 of light intensity signal are penetrated, beam attenuator 38, condenser lenses 12 is detected, is relayed amplifying lens 13 and positioned at relaying
The ccd detector 30 of the focus of amplifying lens 13, also includes being received for the dichroic mirror 25, spectrum of exploring laser light induced breakdown spectroscopy 39
The spectrum investigating system 27 of collection lens 26 and the focal point of spectral collection lens 26, and being used for positioned at the direction of measuring surface normal 29
The ionized sample suction pipe 23 and mass spectrometry detection system 24 of the component of ion body feathers 9 of the detection measurement focal beam spot desorption ionization of object lens 6,
Angle between incident light axis 7 and collection optical axis 10 is 2 θ, and symmetrical with regard to measuring surface normal 29.
The function of main composition is as follows：
There is system 4, focal beam spot to quilt by point source 1, along the collimating lens 2, ring light that the direction of incident light axis 7 is placed
The laser focusing system that the measurement object lens 6 of test sample product 8 are constituted is used to produce the small focal beam spot more than diffraction limit, and this is super to spread out
Penetrate microsize hot spot there is measuring samples surface and the dual-use function of surface plasma is produced.
Put by the collection object lens 11 along collection optical axis 10 direction, condenser lenses 12, relaying amplifying lens 13 and positioned at relaying
The laser dual-axis differential confocal detection system that the ccd detector 30 of the big focal point of lens 13 is constituted carries out precision to sample 8
Focus, and the microcell of sample 8 is focused on to measuring object lens 6 and be imaged, measure the form letter in correspondence focal beam spot region
Breath；
The mass spectrometry detection system being made up of ionized sample suction pipe 23 and mass spectrometry detection system 24 is based on time-of-flight method (TOF)
Detect charge atom, molecule in plasma plume 9 etc. to carry out flight time mass spectrum detection.
The spectral collection lens 26 of light direction are reflected by collection object lens 11, dichroic mirror 25, positioned at dichroic mirror 25 and positioned at light
The spectrum investigating system that the spectrum investigating system 27 of the spectrum focal point of collecting lens 26 is constituted, for luring the laser of sample 8
Lead breakdown spectral 39 to be detected, measure the component information in correspondence focal beam spot region；
Light intensity regulating system is constituted by outgoing beam attenuator 37 and detection beam attenuator 38, for focal beam spot of decaying
The spot intensity detected with ccd detector 30, to adapt to light intensity demand when sample surfaces are positioned
There is the ring light transverse super-resolution system that system 4 and measurement object lens 6 are constituted by ring light, for compression light is focused on
Speckle lateral dimension.
The radial polarisation light longitudinal field being made up of vector beam generation system 31, iris filter 32 and measurement object lens 6 is tight
Focusing system is used to compress focal beam spot lateral dimension.
The three-dimensional motion system being made up of computer 22, three-dimensional working platform 28 can carry out axially focusing to sample 8
Position and 3-D scanning.
The wavelength of pulse laser 33, pulsewidth and repetition rate can be selected as needed.
The process that sample carries out high resolution mass spectrum imaging is mainly included the following steps that：
Step one, the light beam of the outgoing of pulse laser 33 are collimated Jing after collecting lenses 34, Optic transmission fiber 36 and collimating lens 2
For collimated light beam 3, the Jing outgoing beams attenuator 37, ring light of collimated light beam 3 generation generation annular beam 5 of system 4, ring light
Again Jing measurements object lens 6 are focused to be radiated on sample 8 more than the small spot of diffraction limit beam 5；
Step 2, controlled using computer 22 three-dimensional working platform 28 make by the collection object lens 11 along collection optical axis 10 direction,
The laser that condenser lenses 12, relaying amplifying lens 13 and the ccd detector 30 positioned at the relaying focal point of amplifying lens 13 are constituted is double
Axle differential confocal detection system carries out axial scan to sample 8, and to amplifying Airy disk 14 segmentation detection is carried out, and measures correspondence
First off-axis confocal axial strength curve 19 and second of the first microcell of Airy disk 17 and the second microcell of Airy disk 18 is off-axis confocal
Axial strength curve 20；
Step 3, the off-axis confocal axial strength curve 20 of the first off-axis confocal axial strength curve 19 and second is subtracted each other into place
Reason obtains dual-axis differential confocal axial strength curve 21；
The dead-center position z of step 4, computer foundation dual-axis differential confocal axial strength curve 21AThe three-dimensional work of value control
Platform 25 drives sample 8 to move along the direction of measuring surface normal 29, and the focal beam spot for making measurement object lens 6 focuses on sample
On, realize initially focusing to sample 8；
Step 5, the focal beam spot intensity for adjusting outgoing beam attenuator 37 to strengthen measurement object lens 6 make the table of sample 8
Face produces plasma, the plasma plume 9 for producing focal beam spot desorption ionization sample 8 using ionized sample suction pipe 23
In molecule, atom and ion suction mass spectrometry detection system 24 in carry out mass spectrum imaging, measure the matter in correspondence focal beam spot region
Spectrum information；
Step 6, using by the collection object lens 11 along collection optical axis 10 direction, condenser lenses 12, relaying amplifying lens 13
The laser dual-axis differential confocal detection system constituted with the ccd detector 30 positioned at the relaying focal point of amplifying lens 13 is simultaneously to quilt
The corresponding microcell form of 8 surface plasma body feathers of test sample product 9 is imaged, and measures regional morphology information, detects beam attenuator
38 are used to decay light intensity to avoid the supersaturation of ccd detector 30 from detecting；
Step 7, while, Jing dichroic mirrors 25 are reflected using spectrum investigating system 27 and spectral collection lens 26 collect
LIBS 39 carries out light spectrum image-forming detection, measures the component information in correspondence focal beam spot region；
Laser focusing microcell shape information that step 8, computer 22 measure laser dual-axis differential confocal detection system,
The LIBS 39 of the laser focusing microcell that spectrum investigating system 27 is detected simultaneously, mass spectrometry detection system 24 are visited simultaneously
The Information in Mass Spectra of the laser focusing microcell of survey carries out fusion treatment, and form, spectrum and the mass spectrum of focal beam spot microcell are obtained then
Information；
Step 9, the control three-dimensional working platform 28 of computer 22 make measurement object lens 6 be directed at the area next to be measured of sample
Domain, is then operated by step 2~step 7, obtains the form and Information in Mass Spectra of next focal zone to be measured；
The all tested points of step 10, repeat step eight on sample 8 are measured, then using computer 18
Carry out data fusion and image reconstruction process, you 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, point source 1
Can be substituted by the pin hole 35 of the pulse laser 33 along the direction of incident light axis 7, collecting lenses 34, the focal point of collecting lenses 34,
Ring light occurs system 4 can be substituted by vector beam generation system 31, iris filter 32, burnt positioned at relaying amplifying lens 13
The first light intensity point probe 15 and the second light intensity point probe 16 in face is substituted by ccd detector 30.
The radial polarisation light longitudinal field being made up of vector beam generation system 31, iris filter 32 and measurement object lens 6 is tight
Focusing system is used to compress focal beam spot lateral dimension.
Remaining imaging measurement method is same as Example 1.
The specific embodiment of the present invention is described above in association with accompanying drawing, but these explanations can not be understood to limit
The scope of the present invention.
Protection scope of the present invention is limited by appended claims, any changing on the basis of the claims in the present invention
Dynamic is all protection scope of the present invention.
Claims (4)
1. a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum micro imaging method, it is characterised in that：Using high-altitude
Between differentiate dual-axis differential confocal microscopic system focal beam spot sample is focused and imaging, using mass spectrometry detection system to double
Axle differential confocal microscopic system focal beam spot desorption ionization sample and charged molecule, atom for producing etc. carry out microcell mass spectrum into
Picture, dual-axis differential confocal microscopic system focal beam spot desorption ionization sample is produced plasma using spectrum investigating system
Emission spectrum is detected, and the fusion that detection data information is then passed through again realizes that sample microcell is high then with analysis is compared
Spatial discrimination and highly sensitive form and imaging while component and detection, comprise the following steps：
Step one, collimated light beam (3) is set to occur to be shaped as annular beam (5), the annular beam after system (4) by ring light
(5) again Jing measurements object lens (6) focuses on desorption ionization on sample (8) and produces plasma plume (9)；
Step 2, make computer (22) control three-dimensional working platform (28) drive sample (8) along measuring surface normal (29) direction
Move up and down in measurement object lens (6) near focal point, using collection object lens (11), condenser lenses (12), relaying amplifying lens (13)
With positioned at relaying amplifying lens (13) focal plane and with regard to gather optical axis (10) symmetrically placed the first light intensity point probe (15) and
Second light intensity point probe (16) carries out segmentation detection to amplifying Airy disk (14), obtains the microcell of Airy disk first (17) and Airy
The strength characteristics of the microcell of speckle second (18) are respectively the first off-axis confocal axial strength curve (19) and second off-axis confocal
Axial strength curve (20)；
Step 3, the first off-axis confocal axial strength curve (19) and the second off-axis confocal axial strength curve (20) are subtracted each other into place
Reason obtains dual-axis differential confocal axial strength curve (21)；
The dead-center position z of step 4, computer (22) foundation dual-axis differential confocal axial strength curve (21)AThe three-dimensional work of value control
Making platform (28) drives sample (8) to move along measuring surface normal (29) direction, and the focal beam spot for making measurement object lens (6) is focused on
On sample (8)；
Step 5, the plasma plume for being produced focal beam spot desorption ionization sample (8) using ionized sample suction pipe (23)
(9) mass spectrum imaging is carried out in the molecule, atom and ion suction mass spectrometry detection system (24) in, correspondence focal beam spot region is measured
Information in Mass Spectra；
Step 6, using by three-dimensional working platform (28), collection object lens (11), condenser lenses (12), relaying amplifying lens (13) and
Positioned at relaying amplifying lens (13) focal plane and with regard to gathering optical axis (10) symmetrically placed the first light intensity point probe (15) and the
The laser dual-axis differential confocal detection system that two light intensity point probes (16) are constituted focuses on sample to measuring object lens (6)
(8) microcell is imaged, and measures the shape information in correspondence focal beam spot region；
Step 7, Jing colour annalyzers (25) reflection and spectral collection lens (26) are collected laser using spectrum investigating system (27)
Induced breakdown spectroscopy (39) is detected, and measures the spectral information in correspondence focal beam spot region；
Laser focusing microcell shape information, light that step 8, computer (22) measure laser dual-axis differential confocal detection system
Spectrum detection system (27) is while spectral information, the mass spectrometry detection system (24) of the laser focusing microcell of detection is while the laser for measuring
Focusing on the Information in Mass Spectra of microcell carries out fusion treatment, and form, spectrum and the Information in Mass Spectra of focal beam spot microcell are obtained then；
Step 9, computer (22) control three-dimensional working platform (28) make measurement object lens (6) focus be directed at the next of sample (8)
Individual region to be measured, is then operated by step 2~step 8, obtains form, spectrum and the matter of next focal zone to be measured
Spectrum information；
The all tested points of step 10, repeat step nine on sample (8) are measured, then using computer (22)
Processed and be obtained 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：Can be to make collimated light beam (3) that system (31), iris filter (32) occur by vector beam including step one
After be shaped as annular beam (5), again Jing measurements object lens (6) focuses on sample (8) desorption ionization and produces the annular beam (5)
Raw plasma plume (9).
3. a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device, it is characterised in that：Including a light
There is system (4) and gather in source (1), the collimating lens (2) along the placement of incident light axis (7) direction, the ring light of generation annular beam
Burnt hot spot to sample (8) measurement object lens (6), including along collection optical axis (10) direction place for detection measurement object lens
(6) the collection object lens (11) of focal beam spot intensity of reflected light signal, condenser lenses (12), relaying amplifying lens (13) and it is located at
After amplifying lens (13) focal plane and with regard to gathering the first symmetrically placed light intensity point probe (15) of optical axis (10) and the second light intensity
Point probe (16), dichroic mirror (25) also including exploring laser light induced breakdown spectroscopy (39), positioned at dichroic mirror (25) reflected light
The spectral collection lens (26) in direction and the spectrum investigating system (27) positioned at spectral collection lens (26) focal point, and along survey
Amount face normal (29) direction, the ionization sample for detecting ion body feathers (9) component for measuring object lens (6) focal beam spot desorption ionization
Product suction pipe (23) and mass spectrometry detection system (24), the angle between incident light axis (7) and collection optical axis (10) is 2 θ, and with regard to surveying
Amount face normal (29) is symmetrical.
4. a kind of high-space resolution laser dual-axis differential confocal spectrum-mass spectrum microscopic imaging device according to claim 3,
It is characterized in that：System (4) occurs including ring light can be with the arrow of the generation vector beam placed along incident light axis (7) direction
Amount beam production system (31) and iris filter (32) are substituted.
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| CN104931481B (en) * | 2015-06-23 | 2017-08-25 | 北京理工大学 | Laser dual-axis differential confocal induced breakdown Raman spectrum imaging detection method and device |
| CN105067570A (en) * | 2015-07-17 | 2015-11-18 | 北京理工大学 | Dual-axis laser differential confocal LIBS (laser-induced breakdown spectroscopy), RS (Raman spectroscopy) and MS (mass spectrometry) imaging method and device |
| CN105241850A (en) * | 2015-07-17 | 2016-01-13 | 北京理工大学 | Biaxial laser differential confocal LIBS, Raman spectrum-mass spectrum microscopic imaging method and Raman spectrum-mass spectrum microscopic imaging device |
| JP2022500632A (en) * | 2018-09-10 | 2022-01-04 | フリューダイム カナダ インコーポレイテッド | Autofocus sample imaging device and method |
| CN109187720A (en) * | 2018-11-13 | 2019-01-11 | 北京理工大学 | Postposition is divided pupil laser differential confocal mass spectrum micro imaging method and device |
| CN109187719A (en) * | 2018-11-13 | 2019-01-11 | 北京理工大学 | Postposition is divided pupil confocal laser LIBS spectrum-mass spectrum micro imaging method and device |
| CN109187501A (en) * | 2018-11-13 | 2019-01-11 | 北京理工大学 | Postposition is divided pupil laser differential confocal LIBS spectrum micro imaging method and device |
| CN109187718A (en) * | 2018-11-13 | 2019-01-11 | 北京理工大学 | Postposition is divided pupil differential confocal LIBS spectrum-mass spectrum micro imaging method and device |
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| US7423260B2 (en) * | 2005-11-04 | 2008-09-09 | Agilent Technologies, Inc. | Apparatus for combined laser focusing and spot imaging for MALDI |
| JP2008002862A (en) * | 2006-06-21 | 2008-01-10 | Ntp:Kk | Photodetachment analyzer |
| CN103411957B (en) * | 2013-08-22 | 2015-08-12 | 北京理工大学 | High-space resolution twin shaft confocal spectrum micro imaging method and device |
| CN103969239B (en) * | 2013-09-06 | 2016-04-13 | 北京理工大学 | A kind of point pupil laser differential confocal Raman spectra test method and device |
| CN204086141U (en) * | 2014-08-28 | 2015-01-07 | 中国科学院近代物理研究所 | Local laser mass spectrum spectroscopic simultaneous analyzer |
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| CN109254072A (en) * | 2018-11-13 | 2019-01-22 | 北京理工大学 | A kind of laser differential confocal Raman-LIBS- mass spectrometry micro imaging method and device |
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