CN104698070B - High-space resolution confocal laser mass spectrum micro imaging method and device - Google Patents
High-space resolution confocal laser mass spectrum micro imaging method and device Download PDFInfo
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Abstract
The present invention relates to a kind of high-space resolution confocal laser mass spectrum micro imaging method and devices, belong to confocal microscopic imaging technology and mass spectrum imaging technical field.Confocal imaging technology and mass spectrum imaging technology spectrographic detection technology are combined by the present invention, axial fixed-focus and imaging are carried out to sample using the focal beam spot of high-space resolution confocal microscope system, desorption ionization is carried out to sample to carry out mass spectrum imaging using same focal beam spot, and then realizes the high-space resolution imaging of sample microcell image and component.Device includes point light source, collimation lens, generates ring light generating system, spectroscope, mesoporous speculum and middle hole measurement object lens, further include for the light collecting lens of hole measurement object lens focal beam spot intensity of reflected light signal in detection and positioned at the light intensity point probe of light collecting lens focus, and for the ionized sample suction pipe and mass spectrometry detection system of the ion body feathers component of hole measurement object lens focal beam spot desorption ionization in detection.The present invention can be used for the high-resolution imaging of biological mass spectrometry.
Description
Technical field
The invention belongs to confocal microscopic imaging technology and mass spectrum imaging technical field, by confocal laser micro-imaging technique with
Mass spectrum imaging technology is combined, and is related to a kind of high-space resolution confocal laser mass spectrum micro imaging method and device, be can be used for giving birth to
Substance composes the high-resolution imaging of imaging field.
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 focused respectively under the action of electric field and magnetic field obtain by mass-to-charge ratio size order arrange
The collection of illustrative plates instrument of row.Mass spectrum imaging be multiple tiny areas in sample 2 dimensional region are analyzed by mass spectrometry respectively it is specific to detect
The distribution of mass-to-charge ratio (m/z) substance.
From the substance assistant laser desorpted ionized this highly sensitive and high quality detection range life of last century the mid-80
The appearance of substance spectral imaging technology has opened up the brand-new field-biological mass spectrometry of mass-spectrometry one, has promoted mass-spectrometric technique application model
Enclose the various fields for expanding to life science, especially mass spectrum answering in protein, nucleic acid, glycoprotein assay etc.
With not only providing new tool for life science, but also also promote the development of mass-spectrometric technique itself.
But there are problems following prominent for existing substance assistant laser desorpted ionized mass spectrograph:
1) due to being focused come desorption ionization sample using simple laser, thus its there are still laser focal beam spots big, matter
Compose the problems such as space exploration resolving power is not high;
2) long the time required to mass spectrum imaging, laser mass spectrometry instrument focal beam spot axial position often drifts about with respect to sample
Problem.
And the accurate acquisition of biological sample " microcell " Information in Mass Spectra is extremely important for life science.
In fact, how to detect microcell Information in Mass Spectra with sensitivity at present is the important technological problems that biological mass spectrometry field is urgently studied.
The imaging detection mechanism of laser scanning confocal microscopy " point illumination " and " point detection ", not only makes its transverse resolution more same
Etc. parameters light microscope improve 1.4 times, but also make confocal microscope pole be convenient for and super-resolution pupil filtering technique, radial direction
Polarised light tightly focused technology etc. further realizes high-space resolution micro-imaging in conjunction with focal beam spot is compressed.
Based on this, a kind of high-space resolution confocal laser mass spectrum micro imaging method of present invention proposition and device will swash
The detecting function of light confocal microscope focal beam spot focuses desorption ionization function with laser and blends, at through super resolution technology
The small focal beam spot of the confocal microscope of reason carries out high-space resolution imaging to sample, utilizes the same focusing light of confocal microscope
Spot carries out desorption ionization to sample and is imaged for mass spectrometry detection system, then realizes the high-space resolution figure of sample microcell
As imaging and high-space resolution mass spectrum micro-imaging.
A kind of high-space resolution confocal laser mass spectrum micro imaging method of present invention proposition can be that biological mass spectrometry is high with device
Resolution imaging provides a completely new effective technical way.
Invention content
It is focused in imaging process the purpose of the invention is to improve the spatial resolution of mass spectrum micro-imaging technique, inhibit
The drift of hot spot relative sample proposes a kind of high-space resolution confocal laser mass spectrum micro imaging method and device, to simultaneously
Obtain sample component space information and functional information.
The purpose of the present invention is what is be achieved through the following technical solutions.
A kind of high-space resolution confocal laser mass spectrum micro imaging method of the present invention is confocal aobvious using high-space resolution
The focal beam spot of micro-system carries out axial fixed-focus and imaging to sample, utilizes the same focusing of high-space resolution confocal microscope system
Hot spot carries out desorption ionization to carry out mass spectrum imaging to sample, so realize the high-space resolution of sample microcell image and component at
Picture includes the following steps:
Step 1: being shaped as annular beam after so that collimated light beam is passed through ring light generating system, the annular beam is again through dividing
Light microscopic, mesoporous speculum, which are reflected into middle hole measurement object lens and focus on desorption ionization on sample, generates plasma plume;
Step 2: making computer control by middle hole measurement object lens, axial objective scanner, mesoporous speculum, spectroscope, collection
The confocal laser detection system that optical lens and light intensity point probe are constituted carries out axis by axial objective scanner to sample
Confocal axial strength curve is measured to scanning;
Step 3: by confocal axial strength curve along z to translation s after obtain shifting confocal axial strength curve, then will
Shift confocal axial strength curve together focal axis to intensity curve subtract each other processing obtain dislocation subtract each other confocal curves;
Step 4: dislocation to be subtracted each other to the dead-center position z of confocal curvesAIt subtracts shift value s/2 and obtains (zA- s/2), computer according to
According to (zA- s/2) the axial objective scanner of value control makes the focal beam spot of middle hole measurement object lens focus on sample;
Step 5: in the plasma plume generated focal beam spot desorption ionization sample using ionized sample suction pipe
Mass spectrum imaging is carried out in molecule, atom and ion sucking mass spectrometry detection system, measures the Information in Mass Spectra in corresponding focal beam spot region;
Step 6: using by middle hole measurement object lens, axial objective scanner, mesoporous speculum, spectroscope, light collecting lens and
The confocal laser detection system centering hole measurement object lens constituted positioned at the light intensity point probe of laser lens focus focus on tested
The microcell of sample is imaged, and the shape information in corresponding focal beam spot region is measured;
Step 7: the laser that computer measures confocal laser detection system focuses microcell shape information and mass spectrometry detection system
The Information in Mass Spectra that system while the laser measured focus microcell carries out fusion treatment, then obtains the form and matter of focal beam spot microcell
Spectrum information;
Step 8: computer control two-dimentional work bench makes next area to be measured of middle hole measurement object lens alignment sample
Then domain is operated by step 2~step 7, obtain the form and Information in Mass Spectra of next focal zone to be measured;
Measured until all tested points on sample Step 9: repeating step 8, then utilize computer into
Row processing can be obtained sample shape information and Information in Mass Spectra.
In high-space resolution confocal laser mass spectrum micro imaging method of the present invention, including step 1 can be to make collimated light beam
By being shaped as annular beam after vector beam generating system, spectroscope and iris filter, the annular beam is anti-through mesoporous again
Mirror is penetrated to be reflected into middle hole measurement object lens and focus on desorption ionization generation plasma plume on sample.
In high-space resolution confocal laser mass spectrum micro imaging method of the present invention, including step 4 can be computer foundation
The corresponding position z of confocal axial strength curve maximum MBValue makes the focusing of middle hole measurement object lens to control axial objective scanner
Hot spot focuses on sample.
In high-space resolution confocal laser mass spectrum micro imaging method of the present invention, including step 8 can be that computer controls
Two-dimensional scanning mirrors system makes next region to be measured of middle hole measurement object lens alignment sample, then presses step 2~step
Seven are operated, and the form and Information in Mass Spectra of next focal zone to be measured are obtained.
A kind of high-space resolution confocal laser mass spectrum microscopic imaging device of the present invention, including laser point light source system, edge
Collimation lens that light beam direction of travel is placed generates the ring light generating system of annular beam, spectroscope, mesoporous speculum and poly-
Burnt mesoporous speculum the reflected beams further include focusing light for hole measurement object lens in detecting to the middle hole measurement object lens of sample
The light collecting lens of spot intensity of reflected light signal and light intensity point probe positioned at light collecting lens focus, and surveyed for detecting mesoporous
Measure the ionized sample suction pipe and mass spectrometry detection system of the ion body feathers component of object lens focal beam spot desorption ionization.
In a kind of high-space resolution confocal laser mass spectrum microscopic imaging device of the present invention, including ring light generating system can
It is substituted with the vector beam generating system for the generation vector beam placed along optical axis direction and iris filter.
In a kind of high-space resolution confocal laser mass spectrum microscopic imaging device of the present invention, including laser point light source system can
To be made of pulse laser, condenser lens and pin hole positioned at condenser lens focus.
Advantageous effect
The present invention compares prior art, has the following advantages:
1) confocal microscopy with high-space resolution ability is blended with mass spectrometry detection technology, make confocal microscopy at
As the hot spot of system realizes focusing-detection and sample desorption ionization dual function, it can be achieved that the mass spectrographic high spatial mass spectrum of sample microcell
Micro-imaging;
2) zero crossing for subtracting each other confocal curves using dislocation carries out the advance fixed-focus of sample, and minimum focal beam spot is made to focus on sample
Product surface effectively plays confocal system high-altitude, it can be achieved that sample microcell high-space resolution mass spectrometry detection and microcell micro-imaging
Between the potential differentiated;
3) subtract each other confocal curves zero crossing using dislocation and carry out the advance fixed-focus processing of sample, can inhibit existing mass spectrograph because of length
Drifting problem of the focal beam spot with respect to sample in time mass spectrum imaging;
4) size of focal beam spot had not only been had compressed using annular beam imaging, but also provided structure side for mass spectrometry detection
The best fusion in face, can be improved the spatial resolving power of laser mass spectrometry instrument.
Description of the drawings
Fig. 1 is high-space resolution confocal laser mass spectrum micro imaging method schematic diagram;
Fig. 2 is that high-space resolution confocal laser mass spectrum micro imaging method converts schematic diagram 1;
Fig. 3 is high-space resolution confocal laser mass spectrum microscopic imaging device schematic diagram;
Fig. 4 is the high-space resolution confocal laser mass spectrum micro imaging method and installation drawing of Examples 1 and 2;
Fig. 5 is the high-space resolution confocal laser mass spectrum microscopic imaging device figure of embodiment 3 and 4;
Wherein:1- collimated light beams, 2- light intensity point probe, 3- ring lights generating system, 4- annular beams, 5- spectroscopes,
Hole measurement object lens, 8- samples, 9- plasma plumes, 10- computers, 11- axial direction objective scans in 6- mesoporous speculum, 7-
The confocal axial strength curve of device, 12- light collecting lens, 13- detecting pinholes, 14- light intensity detectors, 15-, 16- shift confocal axial direction
Intensity curve, 17- dislocation subtract each other confocal curves, 18- ionized samples suction pipe, 19- mass spectrometry detections system, 20- two-dimentional work bench,
21- vector beams generating system, 22- iris filters, 23- annular beams, 24- laser point light sources system, 25- collimation lenses,
26- pulse lasers, 27- condenser lenses, 28- pin holes, 29- outgoing beams attenuator, 30- detections beam attenuator, 31- two dimensions
Scanning galvanometer system.
Specific implementation mode
The invention will be further described with reference to the accompanying drawings and examples.
Core of the invention method and apparatus is as shown in figs. 1 and 3, wherein by ring light generating system 3 and middle hole measurement object
The ring light transverse super-resolution system that mirror 7 is constituted, for compressing focal beam spot lateral dimension.
As shown in figure 3, being generated after the collimated collimation of object lens 25 of the laser that can be emitted by point light source system 24 shown in FIG. 1
Collimated light beam 1.
As shown in Fig. 2, can be occurred by the ring light in vector beam generating system 21,22 alternate figures 1 of iris filter
System 3, the radial polarisation light longitudinal field being made of vector beam generating system 21, iris filter 22 and middle hole measurement object lens 7
Tightly focused system is for compressing focal beam spot lateral dimension.
Following embodiment is realized on the basis of Fig. 1 and 3.
Embodiment 1
The high-space resolution confocal laser mass spectrum microscopic imaging device of the embodiment of the present invention based on shown in Fig. 4, including by arteries and veins
Laser 26, condenser lens 27 and the laser point light source system 24 constituted positioned at the pin hole 28 of 27 focus of condenser lens are rushed, along light
Collimation lens 25, outgoing beam attenuator 29, ring light generating system 3, spectroscope 5,6 and of mesoporous speculum of axis direction placement
Positioned at the catadioptric direction of optical axis and focus 6 the reflected beams of mesoporous speculum to sample 8 middle hole measurement object lens 7, further include by
Detect beam attenuator 30, light collecting lens 12, positioned at 12 focal point of laser lens light intensity point probe 2 constitute confocal light intensity
Detection system, and 9 component of plasma plume for 7 focal beam spot desorption ionization of hole measurement object lens in detection ionized sample
Suction pipe 18 and mass spectrometry detection system 19.Wherein point probe 2 can be made of detecting pinhole 13 and light intensity detector 14.
By laser point light source system 24 (including pulse laser 26, condenser lens 27 and pin hole 28), collimation lens 25, ring
The laser that shape light generating system 3, spectroscope 5, mesoporous speculum 6, axial objective scanner 11 and middle hole measurement object lens 7 are constituted is poly-
Burnt system is used to generate the small focal beam spot more than diffraction limit, which, which has, measures sample surfaces
With the dual function for generating surface plasma.
By middle hole measurement object lens 7, mesoporous speculum 6, spectroscope 5, light collecting lens 12, detecting pinhole 13 and light intensity detector
The 14 confocal laser detection systems constituted, for carrying out accurate fixed-focus to sample 8 and measuring small focal beam spot region
Form.
Time-of-flight method (TOF) is based on by the mass spectrometry detection system that ionized sample suction pipe 18 and mass spectrometry detection system 19 are constituted
Charge atom, the molecule etc. in plasma plume 9 are detected, to carry out flight time mass spectrum detection.
The three-dimensional motion system being made of computer 10, two-dimentional work bench 20 and axial objective scanner 11 can be to detected sample
Product 8 carry out axial fixed-focus positioning and 3-D scanning.
Light intensity regulating system is constituted by outgoing beam attenuator 29 and detection beam attenuator 30, for focal beam spot of decaying
The intensity that hot spot is detected with light intensity point probe 2, to adapt to light intensity demand when sample surfaces positioning.
Wavelength, pulsewidth and the repetition rate of pulse laser 26 can select as needed.
The process that high resolution mass spectrum imaging is carried out to sample mainly includes the following steps that:
Step 1: collimation is after light beam line focus lens 27, pin hole 28 and collimation lens 25 that pulse laser 26 is emitted
Collimated light beam 1, the collimated light beam 1 through outgoing beam attenuator 29, ring light generating system 3, spectroscope 5, mesoporous speculum 6,
The small spot more than diffraction limit is focused to after middle hole measurement object lens 7 to be radiated on sample 8, desorption ionization generates etc. from
Daughter plumage 9;
Step 2: controlling axial objective scanner 11 using computer 10 makes by middle hole measurement object lens 7, axial objective scan
The confocal laser detection system that device 11, mesoporous speculum 6, spectroscope 5, light collecting lens 12 and light intensity point probe 2 are constituted is to quilt
Sample 8 carries out axial scan, measures confocal axial strength curve 15;
Step 3: by confocal axial strength curve 15 along z to translation s after obtain shifting confocal axial strength curve 16, so
To shift afterwards confocal axial strength curve 16 together focal axis to intensity curve 15 subtract each other processing obtain dislocation subtract each other confocal curves 17;
Step 4: computer 10 subtracts each other the dead-center position z of confocal curves 17 according to dislocationASubtract (the z of translation s/2A-s/2)
Value controls axial objective scanner 11, so that the focal beam spot of middle hole measurement object lens 7 is focused on sample 8, realizes to quilt
The initial fixed-focus of sample 8;
Step 5: adjust outgoing beam attenuator 29 makes detected sample come the focal beam spot intensity of hole measurement object lens 7 in enhancing
8 surface of product generates plasma, the plasma for generating focal beam spot desorption ionization sample 8 using ionized sample suction pipe 18
Mass spectrum imaging is carried out in molecule, atom and ion sucking mass spectrometry detection system 19 in body feathers 9, measures corresponding focal beam spot region
Information in Mass Spectra;
Step 6: using by middle hole measurement object lens 7, axial objective scanner 11, mesoporous speculum 6, spectroscope 5, detection
The confocal laser detection system that beam attenuator 30, light collecting lens 12, detecting pinhole 13, light intensity detector 14 are constituted is simultaneously to quilt
8 surface plasma body feathers of sample, 9 corresponding microcell form is imaged, and regional morphology information is measured, and detects beam attenuator
30 detect for the light intensity that decays to avoid 14 supersaturation of light intensity detector;
Step 7: the laser that computer 10 measures confocal laser detection system focuses microcell shape information and mass spectrometry detection
The laser that system 19 detects simultaneously focuses microcell Information in Mass Spectra and carries out fusion treatment, obtains the form and mass spectrum letter of the focusing microcell
Breath;
Step 8: computer 10, which controls two-dimentional work bench 20, makes the next of middle 7 optical axis of hole measurement object lens alignment sample 8
A region to be measured, is then operated by step 2~step 7, and the form and mass spectrum letter of next focal zone to be measured are obtained
Breath;
Step 9: repetition step 8 is measured until all tested points on sample 8, computer 10 is then utilized
Carry out data fusion and image reconstruction process, you can obtain sample shape information and Information in Mass Spectra.
Embodiment 2
As shown in figure 4, in the high-space resolution confocal laser mass spectrum microscopic imaging device of embodiment 1, computer 10 can
With according to the corresponding position z of 15 maximum value M of confocal axial strength curveBValue makes middle hole measurement to control axial objective scanner 11
The focal beam spot of object lens 7 focuses on sample 8.
Remaining imaging measurement method is same as Example 1.
Embodiment 3
As shown in figure 5, in the high-space resolution confocal laser mass spectrum microscopic imaging device of embodiment 1, ring light occurs
System 3 is substituted with the vector beam generating system 21 for the generation vector beam placed along optical axis direction and iris filter 22, hair
Raw annular beam 23 is focused to be radiated at more than the small spot of diffraction limit after mesoporous speculum 6, middle hole measurement object lens 7
On sample 8.
The radial polarisation light being made of vector beam generating system 21, iris filter 22 and middle hole measurement object lens 7 is longitudinal
Field tightly focused system is for compressing focal beam spot lateral dimension.
Remaining imaging measurement method is same as Example 1.
Embodiment 4
As shown in figure 5, in the high-space resolution confocal laser mass spectrum microscopic imaging device of embodiment 1, computer 10 can
To control next region to be measured that two-dimensional scanning mirrors system 31 makes middle hole measurement object lens 7 be directed at sample 8.
Remaining imaging measurement method is same as Example 3.
The specific implementation mode of the present invention is described above in association with attached drawing, but these explanations cannot 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 (7)
1. a kind of high-space resolution confocal laser mass spectrum micro imaging method, it is characterised in that:It is confocal aobvious using high-space resolution
The focal beam spot of micro-system carries out axial fixed-focus and imaging to sample, utilizes the same focusing of high-space resolution confocal microscope system
Hot spot carries out desorption ionization to carry out mass spectrum imaging to sample, so realize the high-space resolution of sample microcell image and component at
Picture includes the following steps:
Step 1: making collimated light beam (1) by being shaped as annular beam (4), the annular beam after ring light generating system (3)
(4) middle hole measurement object lens (7) are reflected into through spectroscope (5), positioned at the mesoporous speculum (6) of spectroscope (5) transmission direction again
And it focuses on desorption ionization on sample (8) and generates plasma plume (9);
Step 2: making computer (10) control by the axial direction of middle hole measurement object lens (7) and middle hole measurement object lens (7) coaxial placement
Objective scanner (11), mesoporous speculum (6), spectroscope (5), be located at spectroscope (5) reflection direction light collecting lens (12) and
The confocal laser detection system that light intensity detector (2) is constituted carries out axis by axial objective scanner (11) to sample (8)
Confocal axial strength curve (15) is measured to scanning;
Step 3: by confocal axial strength curve (15) along z to translation s after obtain shifting confocal axial strength curve (16), so
After will shift confocal axial strength curve (16) together focal axis to intensity curve (15) subtract each other processing obtain dislocation subtract each other confocal song
Line (17);
Step 4: dislocation to be subtracted each other to the dead-center position z of confocal curves (17)AIt subtracts shift value s/2 and obtains (zA- s/2), computer
(10) according to (zA- s/2) so that the focal beam spot of middle hole measurement object lens (7) is focused on tested for the axial objective scanner (11) of value control
On sample (8);
Step 5: the plasma plume for being generated focal beam spot desorption ionization sample (8) using ionized sample suction pipe (18)
(9) mass spectrum imaging is carried out in the molecule, atom and ion sucking mass spectrometry detection system (19) in, measures corresponding focal beam spot region
Information in Mass Spectra;
Step 6: utilizing the axial objective scanner by middle hole measurement object lens (7) and middle hole measurement object lens (7) coaxial placement
(11), mesoporous speculum (6), spectroscope (5), be located at spectroscope (5) reflection direction light collecting lens (12) and positioned at laser it is saturating
The confocal laser detection system centering hole measurement object lens (7) that the light intensity detector (2) of mirror (12) focus is constituted focus on detected sample
The microcell of product (8) is imaged, and the shape information in corresponding focal beam spot region is measured;
Step 7: the laser that computer (10) measures confocal laser detection system focuses microcell shape information and mass spectrometry detection system
The Information in Mass Spectra that system (19) while the laser measured focus microcell carries out fusion treatment, then obtains the form of focal beam spot microcell
And Information in Mass Spectra;
Step 8: computer (10) control two-dimentional work bench (20) makes the next of middle hole measurement object lens (7) alignment sample (8)
A region to be measured, is then operated by step 2~step 7, and the form and mass spectrum letter of next focal zone to be measured are obtained
Breath;
Step 9: repetition step 8 is measured until all tested points on sample (8), computer (10) is then utilized
It carries out processing and can be obtained sample shape information and Information in Mass Spectra.
2. a kind of high-space resolution confocal laser mass spectrum micro imaging method according to claim 1, it is characterised in that:Packet
Including step 1 and could alternatively be makes collimated light beam (1) pass through vector beam generating system (21), spectroscope (5) and iris filter
(22) annular beam (23) is shaped as after, which is reflected into middle hole measurement object lens through mesoporous speculum (6) again
(7) and desorption ionization generation plasma plume (9) on sample (8) is focused on.
3. a kind of high-space resolution confocal laser mass spectrum micro imaging method according to claim 1, it is characterised in that:Packet
It includes step 4 and could alternatively be computer (10) according to the corresponding position z of confocal axial strength curve (15) maximum value MBValue is controlled
The axial objective scanner (11) of system makes the focal beam spot of middle hole measurement object lens (7) focus on sample (8).
4. a kind of high-space resolution confocal laser mass spectrum micro imaging method according to claim 1, it is characterised in that:Packet
Including step 8 and could alternatively be computer (10) control two-dimensional scanning mirrors system (31) keeps the alignment of middle hole measurement object lens (7) tested
Next region to be measured of sample (8), is then operated by step 2~step 7, and next focal zone to be measured is obtained
Form and Information in Mass Spectra.
5. a kind of implementation of high-space resolution confocal laser mass spectrum micro imaging method described in claim 1, feature
It is:The ring of collimation lens (25), generation annular beam including laser point light source system (24), along the placement of light beam direction of travel
Shape light generating system (3), spectroscope (5), mesoporous speculum (6) and mesoporous speculum (6) the reflected beams are focused to sample
(8) middle hole measurement object lens (7) further include the collection for hole measurement object lens (7) focal beam spot intensity of reflected light signal in detecting
Optical lens (12) and the light intensity point probe (2) for being located at light collecting lens (12) focus, and for hole measurement object lens (7) in detecting
The ionized sample suction pipe (18) and mass spectrometry detection system (19) of plasma plume (9) component of focal beam spot desorption ionization.
6. implementation according to claim 5, it is characterised in that:It can be with along light including ring light generating system (3)
The vector beam generating system (21) and iris filter (22) for the generation vector beam that axis direction is placed substitute.
7. implementation according to claim 5, it is characterised in that:It can be swashed with pulse including laser point light source system (24)
Light device (26), condenser lens (27) and the pin hole (28) positioned at condenser lens (27) focus substitute.
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CN103091299A (en) * | 2013-01-21 | 2013-05-08 | 北京理工大学 | Laser differential confocal map microimaging imaging method and device |
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CN1588157A (en) * | 2004-09-16 | 2005-03-02 | 哈尔滨工业大学 | Shaping circular light three differential confocal microscope |
CN103091299A (en) * | 2013-01-21 | 2013-05-08 | 北京理工大学 | Laser differential confocal map microimaging imaging method and device |
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