CN107167455A - Light splitting pupil laser differential confocal CARS micro-spectrometer method and devices - Google Patents
Light splitting pupil laser differential confocal CARS micro-spectrometer method and devices Download PDFInfo
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Abstract
The invention belongs to microspectrum imaging detection technical field, it is related to a kind of light splitting pupil laser differential confocal CARS micro-spectrometer method and devices.The core concept of the present invention is twin-laser as light source activation Reyleith scanttering light and the CARS light for being loaded with sample spectral characteristic, lossless separation is carried out to Reyleith scanttering light and CARS light using dichroic optical system, wherein Reyleith scanttering light carries out geometry detection and positioning, and CARS light carries out spectrographic detection.The present invention utilizes accurate this corresponding characteristic in light splitting pupil laser differential confocal curve zero crossing and focal position, accurate capture and localized excitation light spot focus position, the spectrographic detection of high-precision geometry detection and high-space resolution is realized, a kind of method and apparatus of achievable sample microcell high-space resolution spectrographic detection are constituted.By combining CARS microtechnics, the time of what is inspired the be loaded with Raman diffused light of sample message is short than traditional spontaneous Raman effect, and energy quick nondestructive is detected to sample.The present invention has the advantages that accurate positioning, high-space resolution, lossless detection, spectral detectivity are high, is that microscopic spectrum detection and dimensional measurement provide a kind of new approach.
Description
Technical field
The invention belongs to microspectrum technical field of imaging, it is related to a kind of light splitting pupil laser differential confocal CARS microspectrums
Method of testing and device, available for microcell anti-Stokes scattering (CARS) spectrum of all kinds of samples of quick detection, can be achieved height
Spatial discrimination is imaged and detection.
Technical background
Light microscope is widely used in biomedical sector and materials science field, and quick with modern science
Development, the requirement to micro-imaging is also imaged from structure imaging turning function.Nineteen ninety, confocal Raman spectra microtechnic into
Work(application, greatly improves the possibility for exploring the specific structural constituent of small items and pattern.It is by confocal microscopy and drawing
Graceful spectral technique is combined, the high-resolution tomography feature for possessing confocal microscopy, has nodestructive testing and spectrum analysis concurrently again
A kind of ability, it has also become technological means of important material structure measurement and analysis, is widely used in physics, chemistry, biological doctor
The fields such as, material science, petrochemical industry, food, medicine, criminal investigation.
Traditional spontaneous Raman scattering imaging technique causes its transmission signal extremely weak due to Raman scattering self character, even if
With the laser excitation of high intensity, the good spectrum picture of a CONT is obtained, the action time grown still is needed very.This length
Time effect limits application of the Raman microscopy in biological field.Coherent based on coherent Raman effect is drawn
Graceful scattering (CARS) process can largely strengthen Raman signal, so as to realize quick detection.Coherent Raman effect is logical
Cross the light for being excited to excite molecule is locked in vibration level, the intensity of the vibration signal that this method is produced and exciting light it is strong
Non-linear relation is spent into, very strong signal, also referred to as coherent nonlinear Raman spectrum can be produced.There is very strong energy to turn for it
Change efficiency, the time for exposure is short, the infringement to sample is also smaller, at the same it scattering have certain directionality, easily with it is miscellaneous
Astigmatism is separated.
The generation of coherent anti-stokes raman scattering (CARS) is a third-order nonlinear optical processes, and it needs pumping
Light, stokes light and detection light.In general, the quantity in order to reduce light source, simplifies process, commonly use pump light and replace detection
Light, the relation between them is as shown in Fig. 2 as pump light (wp) and stokes light (ws) frequency difference and Raman active point
When the vibration frequency of son matches, CARS light w will be inspiredas, wherein was=2wp-ws.The generation process of CARS light is comprising specific
Raman active molecule vibration mode and cause interaction of the molecule from ground state to the incident field of excitation state vibrational transition
Process, its energy level schematic diagram is as shown in Figure 3.Fig. 3 (a) represents resonance Raman and the enhancing of off-resonance single photon to CARS processes
Contribution, Fig. 3 (b) represents that resonance Raman and non resonant two photon strengthen the contribution to CARS processes；Work as wpAnd wsBetween frequency difference with
When the vibration frequency of raman active molecule matches, the signal inspired obtains resonant check；
Traditional CARS microscopies using two single wavelength lasers, can only obtain the spectral information of specific frequency spectrum mostly, and
And tradition CARS microscopies do not emphasize system focus ability, cause actual spectrum detecting location to be often in defocus position.
Even if defocus position can also inspire the Raman spectrum of sample and by the spectrometer detection after pin hole, but intensity can not be reasonable
Characterize the correct spectral signal intensity of the point.In CARS microscopic systems, only when system is accurately focused, optimal sky could be obtained
Between resolving power and best spectroscopic probing capabilities.
Above-mentioned reason limits the ability that CARS microscopic systems detect microscopic spectrum, constrains it in finer microscopic spectrum
Test and the application in analysis occasion.Based on above-mentioned situation, the present invention proposes the strong of the sample surfaces scattering for arriving systematic collection
In sample Raman diffused light 103~106Reyleith scanttering light again carries out detected with high accuracy, it is organically blended with spectrum investigating system, enters
The synchronizing detection of row spatial positional information and spectral information, to realize high-space resolution, high spectral resolution light splitting pupil laser
Differential confocal CARS collection of illustrative plates is imaged and detected.
The core concept of patent of the present invention is as sharp from super continuous spectrums pulse laser and unicast long-pulse laser
Light emitting source, expands excitation spectrum scope, improves excitation of spectra intensity；To receiving Reyleith scanttering light light splitting pupil laser differential confocal detection system
System, can make the axial response characteristic curve of light splitting pupil confocal microscope system produce the spy of phase shift using detector lateral shift
Property, symmetry division is carried out by focal spot, differential processing is carried out to two paths of signals, differential detection is realized.It is real according to differential principle
Existing bipolarity actual zero point tracking measurement, is accurately focused, and then realize high-space resolution；After accurately focusing, spectrographic detection is carried out,
Obtain optimal spectrum resolution capability.
The content of the invention
The purpose of the present invention is to overcome the deficiencies in the prior art, propose a kind of light splitting pupil laser differential of high-space resolution
Confocal CARS micro-spectrometers method and its device.
The present invention is achieved by the following technical solutions.Light splitting pupil laser differential confocal CARS micro-spectrometer sides
Method, comprises the following steps：
A) super continuous spectrums laser is sent by super continuous spectrums laser, after bandpass filter by the first dichroscope with
The Single wavelength laser that single wavelength laser is sent converges, and makes that two light beams sequential is consistent, space coincidence (unicast by adjusting light path
Long laser envelope is in continuous spectrum laser)；Mixed light beam is focused on sample by illumination iris and microcobjective, is inspired
Reyleith scanttering light and the CARS light for being loaded with sample spectral characteristic；
B) CARS light and Reyleith scanttering light are divided into two beams after the second dichroscope, wherein the light beam comprising CARS light enters light
Probe unit is composed, light beam of another beam comprising Reyleith scanttering light enters light splitting pupil laser differential confocal probe unit；In spectrographic detection list
In member, the light beam comprising CARS light first passes through bandpass filter, filters out the non-CARS interference light in light beam, then passes through the first meeting
Poly- mirror is assembled by the first pin hole, is filtered and is entered spectrometer by the convergence of the second convergent mirror again after ambient light, obtains CARS spectrum letter
Breath；The light beam of another Shu Zaiyou Reyleith scanttering lights is detected by carrying out focal spot segmentation by intensity collection system after the 3rd convergent mirror, respectively
Obtain the signal corresponding to search coverage A and search coverage B.
C) signal of two search coverages of A, B is carried out it is differential subtract each other after processing, obtain differential confocal curve, utilize light splitting
" zero crossing " of pupil laser differential confocal response curve is touched with measuring objective focus positions accurately corresponding characteristic by " zero crossing "
Send accurate capture and excite hot spot focal position, realize the geometry detection and space orientation of high-space resolution.
D) sample surface location (exciting light spot focus) is obtained by computer disposal, by controlling high-precision three-dimensional
Translation stage movement is scanned, makes Laser Focusing on sample surface, the CARS spectrum for obtaining the point by spectrographic detection unit are believed
Breath.
E) signal of Reyleith scanttering light is individually handled, the three dimension scale tomography of high-space resolution can be achieved；Individually processing
CARS spectral signals, can obtain spectrum picture；Rayleigh signal and CARS spectral signals are handled simultaneously, and high-space resolution can be achieved
Collection of illustrative plates tomography, that is, realize the collection of illustrative plates imaging and detection of sample geometric position information and spectral information.
Particularly, in the methods of the invention, illumination iris and collection pupil can be circular, D-shaped or other shapes.
Particularly, in the methods of the invention, excitation beam includes the light beams such as line polarisation, rotatory polarization, radial polarisation light
With the structure light beam generated by technologies such as pupil filterings, system spectrum Signal-to-Noise and system lateral resolution are thus improved.
Particularly, in the methods of the invention, by matching the optical filter of different spectral bands, the Stokes of different spectral coverage is selected
Light, it is possible to achieve the spectrographic detection of different spectral coverage；Wherein, the optical filtering bands of a spectrum of bandpass filter and bandpass filter are on Single wavelength
The centre wavelength of laser is symmetrical.
Particularly, in the methods of the invention, laser emission element can also add photonic crystal fiber with single wavelength laser
Spectrum widening realization is carried out, in addition, the spectrometer in spectrographic detection unit is substituted for into photoelectricity point probe, rotatory polarization piece can
To realize that spectral scan is exported, and then excite CARS spectrum and CARS spectral signals are obtained by the detection of photoelectricity point probe；
Include the invention provides a kind of light splitting pupil laser differential confocal CARS micro-spectrometer devices：Excitation of spectra list
Member, the laser emission element positioned at excitation of spectra unit incident direction, the dichroic list positioned at excitation of spectra unit exit direction
Member, the spectrographic detection unit positioned at dichroic unit transmission direction, the differential confocal positioned at dichroic unit reflection direction are detected
Unit, the computer for controlling whole system.Laser emission element by unicast long-pulse laser, super continuous spectrums pulse laser,
Bandpass filter and the first dichroscope composition；Excitation of spectra unit by illumination iris, collect pupil, measure object lens, treat test sample
Product and high-precision three-dimensional scanning translation stage composition；Dichroic unit is the second dichroscope；Spectrographic detection unit is by bandpass filter
Piece, the first convergent mirror, the first pin hole, the second convergent mirror and spectrometer composition；Laser differential confocal probe unit is assembled by the 3rd
Mirror and intensity collection system composition.
In apparatus of the present invention, the method that intensity collection system can be combined using double needle hole with dual-quadrant detector is real
Now the segmentation to Airy disc is detected.
In apparatus of the present invention, intensity collection system can use ccd detector, be detected by being set in CCD detection face
The position in region and size, realize that the segmentation to Airy disc is detected.
In apparatus of the present invention, intensity collection system can using conduction optical fiber, by the focal plane of the 3rd convergent mirror,
On symmetrically placed two optical fiber of optical axis, realize that the segmentation to Airy disc is detected.
In apparatus of the present invention, the Aili spot that intensity collection system is detected can be amplified by increasing relaying amplifying lens,
To improve the acquisition precision of light splitting pupil laser differential confocal measurement apparatus.
Beneficial effect
The inventive method, contrast prior art has following innovative point：
1st, the present invention organically combines light splitting pupil laser differential confocal microtechnic and CARS spectrographic detections technology, using dividing
Pupil laser differential confocal curve " zero point " position and the accurate corresponding characteristic in microcobjective focal position, accurately catch exciting light
Spot focal position and detecting light spectrum information, so as to realize spectrum tomography and the detection of high-space resolution.
2nd, using dichroscope is by Reyleith scanttering light and is loaded with the CARS light of sample message and is separated in the present invention, wherein Rayleigh
Light realizes that geometric position is detected into light splitting pupil laser differential confocal probe unit, and CARS light is realized into spectrographic detection unit
CARS spectrographic detections, the spectral information of light spot focus is excited by precisely focusing capture, system spectrum detectivity is improved.
In addition, dichroic light-dividing device can adjustment angle as needed, facilitate structure to debug.
3rd, the present invention by light splitting pupil laser differential confocal microscopic system and CARS spectrum imaging systems structurally and functionally
Upper combination, can be achieved the tomography of sample microcell geometric parameter, the spectrographic detection of sample microcell can be realized again, figure can be also realized
Compose tomography.
The inventive method, contrast prior art has following remarkable advantage：
1st, the light splitting pupil laser differential confocal detection mode that the present invention is used, using the differential characteristic for subtracting each other noise cancellation,
Environment resistant interference performance is strong.
2nd, the present invention carries out high accuracy positioning using light splitting pupil laser differential confocal technology to measurement focal beam spot, and focuses
Point position carries out real-time follow-up, eliminates the ambient influnence such as temperature and disturbance, and realizing the detection of CARS spectrums, accurate correspondence is most all the time
The small sample spectra for exciting focal beam spot region, greatly improves the microscopical microscopic spectrum detectivities of existing CARS and geometry position
Put detectivity.
3rd, the present invention is used as excitation source using super continuous laser matching Single wavelength laser, it is possible to achieve broad band CARS light
Spectrum detection.
Brief description of the drawings
Fig. 1 is Figure of abstract, i.e., the present invention is basic implements figure；
Fig. 2 is coherent (CARS) light excitation principle figure；
Fig. 3 is CARS light and pump light, the graph of a relation of stokes light
Fig. 4 is that conventional polarization detects micro- index path；
Fig. 5 is light splitting pupil laser differential confocal response curve；
Fig. 6 is the micro- method of testing schematic diagrames of D-shaped light splitting pupil laser differential confocal CARS；
Fig. 7 is the micro- method of testing schematic diagrames of light splitting pupil laser differential confocal CARS for combining iris filter；
Fig. 8 is to be illustrated using the micro- method of testings of light splitting pupil laser differential confocal CARS of double needle hole and dual-quadrant detector
Figure；
Fig. 9 is the micro- method of testing schematic diagrames of light splitting pupil laser differential confocal CARS using ccd detector；
Figure 10 is the micro- method of testing schematic diagrames of light splitting pupil laser differential confocal CARS detected using optical fiber；
Figure 11 is the light splitting pupil laser differential confocal CARS microscopic test device schematic diagrames with detection focal spot amplification system；
Figure 12 is the micro- method of testing schematic diagrames of light splitting pupil laser differential confocal CARS of single laser light source；
Figure 13 is the high-space resolution light splitting micro- method of testings of pupil laser differential confocal CARS and schematic device, that is, is implemented
Example is schemed.
In figure, 1- laser emission elements, 2- Single wavelengths LASER Light Source, 3- super continuous spectrums LASER Light Source, 4- bandpass filters,
The dichroscopes of 5- first, 6- measurements object lens, 7- illumination iris, 8- collect pupil, 9- samples, the scanning of 10- high-precision three-dimensionals
Translation stage, the dichroscopes of 11- second, 12- bandpass filters, the condensers of 13- first, the pin holes of 14- first, 15- second condenser lens,
16- spectrometers, 17- spectrographic detections unit, 18- light splitting pupil laser differential confocals probe unit, the condensers of 19- the 3rd, 20- light intensity
The off-axis confocal axial curve of acquisition system, 21- search coverages A curve, 22- search coverages B curve, 23- first, 24- the
Two off-axis confocal axial curve, 25- non-cumulative curves, 26- iris filters, 27- double needle holes, 28- dual-quadrant detectors, 29-CCD
Detector, 30- conduction optical fiber 1,31- conduction optical fiber 2,32- relaying amplifying lens, 33- polarization splitting prisms, 34- polarizers,
35- photonic crystal fibers, the speculums of 36- first, 37- optical time delay line, the speculums of 38- second, 39- photoelectricity point probe, 40-
Computer；
Embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples.
Fig. 1 is light splitting pupil laser differential confocal CARS microspectrum detection method schematic diagrames.First, from a Single wavelength
Pulse laser (2) sends pump light (detection light) as pump light source and probe source by it, then from a frequency with
Consistent super continuous spectrums pulse laser (3) as Stokes light source, obtain requiring ripple after bandpass filter (4)
Continuous spectrum laser in long scope, by adjusting optical texture, makes two beam laser by the first dichroscope (5) time consistency,
Space coincidence；Mixed light is focused on sample (9) by illumination iris (7) and measurement object lens (6)；Due to tightly gathering
In the case of Jiao, the condition of phase matched is easily met, and is inspired Reyleith scanttering light and is loaded with the CARS light of sample spectral characteristic；
CARS light and Reyleith scanttering light arrive separately at spectrographic detection unit (17) after the second dichroscope (11) and light splitting pupil laser differential is common
Burnt probe unit (18)；Wherein, spectrographic detection unit (17) carries out spectrographic detection to CARS light, and light splitting pupil laser differential confocal is visited
Survey unit (18) and geometric position detection is carried out to Reyleith scanttering light.
In light splitting pupil laser differential confocal probe unit (18), Reyleith scanttering light passes through the convergence of the 3rd convergent mirror, by light intensity
Acquisition system carries out segmentation detection, obtains the microcell of Airy disc first and Airy disc the second microcell strength characteristics, i.e., first from
The confocal axial curve (23) of axle and the second off-axis confocal axial curve (24)；By the first off-axis confocal axial curve (23) and second
Off-axis confocal axial curve (24), which is done, subtracts each other processing, that is, obtains light splitting pupil laser differential confocal curve (25), i.e. Fig. 5.
Circular illumination pupil (7) and collection pupil (8) are replaced with into other shapes, such as D-shaped constitutes D-shaped light splitting pupil and swash
Light differential confocal CARS method of testings, as shown in Figure 6.
The spatial resolution of spectrographic detection is improved by adding iris filter (26), that is, constitutes addition iris filter
Light splitting pupil laser differential confocal CARS method of testings, as shown in Figure 7.
The method that intensity collection system (20) can be combined using double needle hole (27) with dual-quadrant detector (28), realization pair
The segmentation detection of Airy disc, as shown in Figure 8.
Intensity collection system (20) can use ccd detector (29), set small on detection focal plane by changing
The parameter in region realizes that the segmentation to Airy disc is detected to match the reflectivity of different samples, so as to extend its application
Field, as shown in Figure 9.
Intensity collection system can be symmetrical along optical axis by the focal plane of the 3rd convergent mirror (19) using conduction optical fiber
Two optical fiber (30,31) are placed, realize that the segmentation to Airy disc is detected, as shown in Figure 10.
Amplification system (32) can be increased in light splitting pupil laser differential confocal detection system, it is poor to improve light splitting pupil laser
The acquisition precision of dynamic confocal measuring apparatus, as shown in figure 11.
Figure 12 is single micro- method of testing schematic diagrames of laser device laser differential confocal CARS, and effect is by laser emission element
Twin-laser input be changed to the input of single laser, reduce cost；Unicast long-pulse laser sends Single wavelength laser, through polarization
Amici prism (33) light splitting, transmissive portion enters photonic crystal fiber (35) progress widening by polarizer (34) and uses band
Pass filter piece carries out particular requirement wavelength interception, and the speculum of reflecting part first (36), optical time delay line (37) and second are anti-
Penetrate mirror (38) to be coupled at the first dichroscope (5) place with the continuous spectrum laser after broadening afterwards, output space is consistent, the time one
The mixed light beam of cause, the CARS excitations of spectra are carried out to sample.Wherein, the effect of optical time delay line (37) is to ensure that two beams swash
Light sequential is overlapped.The spectrometer (16) in spectrographic detection unit (17) is further substituted for photoelectricity point probe (39), rotated
Polarizer (34), which changes light polarization, makes the spectral line of photonic crystal fiber (37) output wavelength consecutive variations as Stokes
Light, and then realize the CARS spectral measurements of broad band.
Embodiment
In the present embodiment, the picosecond laser that wavelength is 532nm is used as pump light source and probe source, using heavy
Consistent super continuous spectrums picosecond laser adds 550~650nmm bandpass filters as Stokes light source to complex frequency therewith,
Outgoing is mixed under conditions of space coincidence, time consistency is met, is obtained by iris filter after structure light beam, illumination of recording a demerit
Pupil and measurement object lens tightly focused now meet phase-matching condition, inspire wave-length coverage in 450~515nm on sample
Anti-Stokes light (CARS) and wavelength be 532nm Reyleith scanttering light.
As shown in figure 13, it is light splitting pupil laser differential confocal CARS micro-spectrometer devices, its testing procedure is as follows：
First, in laser emission element (1), the continuous spectrum laser that super continuous spectrums laser (3) is sent is filtered by band logical
Mating plate (4) obtains 550~650 broad band laser after filtering, then send one-wavelength laser with Single wavelength (532nm) laser (2)
Converge at the first dichroscope (5) place, form mixed light beam, wherein, this two beams laser repetition rate is consistent, reach the one or two to
Look mirror (5) time consistency, light beam can be completely superposed (the complete envelope of pump spot is in Stokes hot spot) after converging；Mixed light
Beam passes through iris filter, produces structure light beam；Structure light beam is by illumination iris with measurement object lens tightly focused in sample
(9) on, inspire Reyleith scanttering light and be loaded with the CARS light of sample (9) spectral characteristic.
Now, Sample Scan can be accomplished by the following way：X-y-z is realized by high-precision three-dimensional scanning translation stage (10)
Vibration mirror scanning structure is added in scanning direction, or light path after laser emitting, x-y scanning directions are realized, and realize by PZT
Axial scan.
The light beam that sample is reflected back includes stokes light λs, pump light λ0, Reyleith scanttering light λ0, CARS light λas；Wherein,
CARS light λasWith stokes light λsInto spectrographic detection unit (17), 532nm pump light and Reyleith scanttering light is through the second dichroic
Mirror (11) is reflected into light splitting pupil laser differential confocal probe unit (18).In spectrographic detection unit (17), by Stokes
Light λsWith CARS light λasThe bandpass filter (12) that the light of mixing first passes through 450~515nm only retains CARS light afterwards, then passes through
First convergent mirror (13) is assembled by the first pin hole (14), is filtered and is entered spectrum by the second convergent mirror (15) convergence again after ambient light
Instrument (16), CARS spectrum I (λ) is obtained so as to detect, and wherein λ is inspired the ripple of CARS light by sample (9) stimulated luminescence
It is long.In light splitting pupil laser differential confocal probe unit (18), Reyleith scanttering light λ0Assembled by the 3rd convergent mirror, by ccd detector
(29) received, by the way that on CCD detection face, position and the size of search coverage are symmetrical arranged along optical axis, Airy disc is carried out
Segmentation detection, obtains the first off-axis confocal axial curve (23) and the second off-axis confocal axial curve (24).Two paths of signals is done
Difference processing, obtains differential wave (25)." zero point " of light splitting pupil laser differential confocal response curve (25) is poly- with excitation beam
Focus is accurately corresponded to, and the elevation information on sample (9) surface is obtained by " zero point " of response curve (25), with reference to connection
The positional information of the displacement transducer feedback of high-precision three-dimensional scanning translation stage (10) reconstructs detected sample by computer (40)
The three-dimensional surface shape I (x, y, z) of product (9).
Computer (40) control high-precision three-dimensional scanning translation stage (10), makes laser vernier focusing in sample (9) table
Face, the CARS spectrum I (λ) of sample spectral characteristic can correctly be characterized by inspiring, and gathered, passed through by spectrographic detection unit (17)
Computer (40) merges positional information I (x, y, z) and spectral information I (λ), completes the three-dimensionalreconstruction and light of sample (9)
Spectrum information fusion I (x, y, z, r).In addition, computer system is also extended through in whole system, computer (40) is used to realize to high-precision
Spend the Bit andits control of 3-D scanning translation stage, the acquisition process of light splitting pupil laser differential confocal signal and CARS spectral signals and
Data Fusion.
More than, along laser emitting direction, be sequentially placed laser emission element (1), illumination iris (7), microcobjective (6),
Sample (9), high-precision three-dimensional translation stage (10), collect pupil (8), the second dichroscope (11), in the second dichroscope
(11) transmission direction places spectrographic detection unit (17), and light splitting pupil laser differential confocal is placed in dichroscope (11) reflection direction
Probe unit (18).Laser emission element (1) includes super continuous spectrums laser (3), bandpass filter (4) and the first dichroscope
And unicast long-pulse laser (2) (5)；Bandpass filter (12), the first convergence are sequentially placed in spectrographic detection unit (17)
Mirror (13), the first pin hole (14), the second convergent mirror (15), spectrometer (16)；In light splitting pupil laser differential confocal probe unit
(18) in, the 3rd convergent mirror (19) and ccd detector (29) are placed respectively.In the entire system, unicast long-pulse laser
(2), super continuous spectrums laser (3), high-precision three-dimensional scanning translation stage (10), spectrometer (16), ccd detector (29) are counted
Calculation machine (40) is controlled, and the three dimensional local information and spectral information that system is obtained also carry out fusion treatment by computer (40).
The embodiment of the present invention is described above in association with accompanying drawing, but these explanations can not be understood to limitation
The scope of the present invention, protection scope of the present invention is limited by appended claims, any in the claims in the present invention base
The change carried out on plinth is all protection scope of the present invention.
Claims (10)
1. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer method and devices, it is characterised in that including following step
Suddenly：
A) super continuous spectrums laser is sent by super continuous spectrums laser (3), and the first dichroscope is passed through after bandpass filter (4)
(5) the Single wavelength laser sent with single wavelength laser (2) converges, and makes that two light beams sequential is consistent, space by adjusting light path
Overlap (Single wavelength laser envelope is in continuous spectrum laser)；Mixed light beam by illumination iris (7) and microcobjective (6) be focused at by
On test sample product (9), inspire Reyleith scanttering light and be loaded with the CARS light of sample (9) spectral characteristic；
B) CARS light and Reyleith scanttering light are divided into two beams after microcobjective, collection pupil (8) and the second dichroscope (11), wherein
Light beam comprising CARS light enters spectrographic detection unit (17), and light beam of another beam comprising Reyleith scanttering light enters light splitting pupil laser differential
Confocal detection unit (18)；In spectrographic detection unit (17), the light beam comprising CARS light first passes through bandpass filter (12), filter
Except the non-CARS in light beam disturbs light, then assembled by the first convergent mirror (13), ambient light is blocked by the first pin hole (14), subtracted
Assembled again by the second convergent mirror (15) after the interference of subenvironment light and enter spectrometer (16), obtain CARS spectral informations；Another beam is carried
After the light beam for having Reyleith scanttering light is assembled by the 3rd convergent mirror (19), received by intensity collection system (20), focal spot is split,
Respectively obtain the signal corresponding to search coverage A (21) and search coverage B (22).
C) after subtracting each other survey region A (21) signals and search coverage B (22) signal normalization, differential confocal curve (25) is obtained,
Using " zero crossing " of light splitting pupil laser differential confocal response curve with measuring objective focus positions accurately corresponding characteristic, pass through
" zero crossing " triggering carrys out accurate capture and excites hot spot focal position, realizes the geometry detection and space orientation of high-space resolution.
D) handled by computer (40) and obtain sample surface location (exciting light spot focus), control high-precision three-dimensional scanning
Translation stage (10) drives sample (9) mobile, makes Laser Focusing on sample (9), passes through spectrographic detection unit (17)
Obtain the CARS spectral informations of the point.
E) signal of Reyleith scanttering light is individually handled, the three dimension scale tomography of high-space resolution can be achieved；Individually handle CARS light
Spectrum signal, can obtain spectrum picture；Rayleigh signal and CARS spectral signals are handled simultaneously, and the collection of illustrative plates layer of high-space resolution can be achieved
Analysis imaging, that is, realize the collection of illustrative plates imaging and detection of sample geometric position information and spectral information.
2. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer methods according to right 1, it is characterised in that shine
Mingguang City's pupil (7) and collection pupil (8) can be circular, D-shaped or other shapes.
3. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer methods according to right 1, it is characterised in that swash
Luminous beam includes the light beams such as line polarisation, rotatory polarization, radial polarisation light and the structure light beam generated by technologies such as pupil filterings,
Thus system spectrum Signal-to-Noise and system lateral resolution are improved.
4. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer methods according to right 1, it is characterised in that logical
The optical filter of overmatching different spectral bands, selects the stokes light of different spectral coverage, it is possible to achieve the spectrographic detection of different spectral coverage；Its
In, the optical filtering bands of a spectrum of bandpass filter (4) and bandpass filter (12) are symmetrical on the centre wavelength of single wavelength laser (2).
5. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer methods according to right 1, it is characterised in that swash
Optical Transmit Unit (1) can also carry out spectrum widening realization with single wavelength laser (2) plus photonic crystal fiber (35), in addition,
Spectrometer in spectrographic detection unit is substituted for photoelectricity point probe (39), rotatory polarization piece (34) can realize spectral scan
Output, and then excite CARS spectrum and CARS spectral signals are obtained by the detection of photoelectricity point probe.
6. a kind of differential confocal CARS micro-spectrometer devices include：It is excitation of spectra unit, incident positioned at excitation of spectra unit
The laser emission element (1) in direction, the dichroic unit (11) positioned at excitation of spectra unit exit direction, positioned at dichroic unit
(11) the spectrographic detection unit (17) of transmission direction, the light splitting pupil laser differential confocal positioned at dichroic unit (11) reflection direction
Probe unit (18), the computer (40) for controlling whole system.Laser emission element (1) is by unicast long-pulse laser (2), super
Continuous spectrum pulse laser (3), bandpass filter (4) and dichroscope (5) composition；Excitation of spectra unit by illumination iris (7),
Collect pupil (8), microcobjective (6), testing sample (9) and high-precision three-dimensional scanning translation stage (10) composition；Dichroic unit
(11) it is the second dichroscope (11)；Spectrographic detection unit (17) is by bandpass filter (12), the first convergent mirror (13), the first pin
Hole (14), the second convergent mirror (15) and spectrometer (16) composition；Light splitting pupil laser differential confocal probe unit (18) is by the 3rd meeting
Poly- mirror (19), intensity collection system (20) composition.
7. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer devices as claimed in claim 6, it is characterised in that
The method that intensity collection system (20) can be combined using double needle hole (27) with dual-quadrant detector (28), is realized to Airy disc
Segmentation detection.
8. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer devices as claimed in claim 6, it is characterised in that
Intensity collection system (20) can use ccd detector (29), by the position of CCD detection face setting search coverage and greatly
It is small, realize that the segmentation to Airy disc is detected.
9. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer devices as claimed in claim 6, it is characterised in that
Intensity collection system (20) can be using conduction optical fiber (30,31), by the focal plane of the 3rd convergent mirror (19), on optical axis
Symmetrically placed two optical fiber, realizes that the segmentation to Airy disc is detected.
10. a kind of light splitting pupil laser differential confocal CARS micro-spectrometer devices as claimed in claim 6, its feature exists
In, can be by increasing image enhancement system (32), the Aili spot that amplification intensity collection system is detected, to improve light splitting pupil laser
The acquisition precision of differential confocal measuring device.
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