CN109211873A - Postposition is divided pupil laser differential confocal Raman spectra test method and device - Google Patents

Postposition is divided pupil laser differential confocal Raman spectra test method and device Download PDF

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CN109211873A
CN109211873A CN201811343472.1A CN201811343472A CN109211873A CN 109211873 A CN109211873 A CN 109211873A CN 201811343472 A CN201811343472 A CN 201811343472A CN 109211873 A CN109211873 A CN 109211873A
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light
differential confocal
raman
pupil
sample
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赵维谦
吴寒旭
邱丽荣
王允
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Beijing Institute of Technology BIT
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    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/65Raman scattering
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01Arrangements or apparatus for facilitating the optical investigation

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Abstract

The present invention relates to postposition light splitting pupil laser differential confocal Raman spectra test method and devices, belong to microspectrum technical field of imaging.The present invention realizes the high-space resolution detection of sample geometrical morphology by the Rayleigh scattering light building light splitting pupil differential confocal microscopic imaging system abandoned in confocal Raman spectra detection system;The Raman spectral information of sample focal point is accurately obtained using the focal position that light splitting pupil differential confocal microscopic imaging system obtains, and then realizes light splitting pupil differential confocal Raman spectra high-space resolution imaging and the detection of " collection of illustrative plates ".The present invention has many advantages, such as registration, and high-space resolution and spectral detectivity are high, can be incorporated in extensively biomedicine, physical chemistry, accurate measurement, in femtosecond laser processing and other fields.

Description

Postposition is divided pupil laser differential confocal Raman spectra test method and device
Technical field
The present invention relates to postposition light splitting pupil laser differential confocal Raman spectra test method and device, belong to microspectrum at As technical field.The present invention combines postposition light splitting pupil laser differential confocal microtechnic with Raman spectroscopic detection technology, relates to And the postposition light splitting pupil laser differential confocal Raman spectra test method and device of a kind of " collection of illustrative plates ", it can be used for all kinds of samples The raman microspectroscopy spectrum of product carries out high-space resolution detection etc..
Background technique
Confocal laser Raman spectroscopy makes to scatter light by the way that energy exchange occurs between incident laser and optical phonon Frequency displacement changes, and realizes the non-cpntact measurement to parameters such as the constituent of sample microcell, concentration, stress, strain, temperature, Confocal laser Raman spectroscopy is also referred to as molecular probe technology.The technology has the high-space resolution layer of confocal microscopy Analysis ability carries out tomoscan by sample, obtains the Raman spectral information of sample different depth, and then non-contacting High-space resolution measurement is carried out to sample microcell under mode.Confocal laser Raman spectroscopy is as a kind of extremely important sample The technological means of composition measurement and analysis, in biomedicine, physical chemistry, material science, environment measuring, food safety, drug Have in the fields such as detection, criminal investigation and is widely applied.
The detection principle of traditional confocal Raman spectra system is as shown in Figure 1, light-source system shoot laser penetrates light splitting rib After mirror, quarter-wave plate and measurement object lens, sample surfaces are focused on, the Raman for being loaded with sample microscopic spectrum characteristic is inspired Light is scattered, by the mobile sample of 3 D scanning system, makes Raman scattering corresponding to different tested regions on sample Light is collected by object lens, and by the reflection of quarter-wave plate and Amici prism, the first convergent mirror is by the Raman scattering of reflection Light converges at the first pin hole, and the Raman spectrum for being loaded with sample microcell characterisitic parameter information is measured using spectrum investigating system.
Traditional confocal Raman spectra system is the signal-to-noise ratio of raising system, obtains raman microspectroscopy spectral information abundant, It usually requires to carry out prolonged Raman spectrum imaging to sample.But instrument in prolonged imaging process be easy by The influence of many factors such as environment temperature, air agitation, system vibration, easily make system generate drift, defocus phenomena such as.Due to The ability of Focus tracking is not present in existing confocal Raman spectra detection system, therefore exciting light is difficult to ensure in imaging process Spot is overlapped with objective focus positions, leads to that hot spot is excited to be greater than ideal focal beam spot, result reduces spectrum investigating system Spatial resolution, therefore limit the detectivity of confocal Raman spectrometer.
Since spontaneous Raman scattering light is very faint, only the 10 of Rayleigh scattering light-3~10-6Times, therefore drawn to reduce Graceful energy loss improves the detected intensity of Raman spectrum, the pin hole of larger size is generallyd use in system, about 150 μm~ Between 200 μm.Since system carries out axially position using confocal focal position, the oversized of pin hole will lead to axial fixed-focus energy The reduction of power, and then reduce the spatial resolution of system.With the development of modern science and technology, people are for crs technique Spatial resolution and spectroscopic probing capabilities more stringent requirements are proposed.In confocal Raman spectra system, work as focal beam spot When size minimum, the excitation intensity of Raman spectrum reaches maximum, and spectroscopic probing capabilities and spatial resolution reach most preferably, therefore essence Determine that coke is of great significance for confocal Raman spectra system.
In addition, faint Raman diffused light is only utilized in traditional confocal Raman spectra system, and intensity is abandoned much Higher than the Rayleigh scattering light of Raman diffused light, therefore spectrographic detection can only be carried out, more sample messages, measurement side can not be obtained Formula is single, limits its application field.And axial fixed-focus and light spectrum image-forming directly are carried out using Raman diffused light, there is survey Measure the disadvantages such as the time is long, system sensitivity is low.
For the above-mentioned deficiency for overcoming traditional confocal Raman system, Beijing Institute of Technology Zhao Wei is modest et al. it is proposed that one kind Light splitting pupil differential confocal Raman spectra test method with microscopic spectrum detectivity, by being split place to detection focal spot Reason, obtains the light intensity signal of two search coverages, carries out subtracting each other processing realization differential detection to it, and then realize that bipolarity is absolute Zero-tracking measurement.Pupil differential confocal Raman spectra measuring technology is divided in (the inventor: Zhao Wei of patent ZL 201410083285 It is modest etc.) in, it is disclosed, principle such as Fig. 2 institute with entitled " light splitting pupil laser differential confocal Raman spectra test method and device " Show, this method is intended to accurately correspond to this spy with its focus using " zero crossing " of light splitting pupil differential confocal microscopic imaging device Property, it realizes and the high-space resolution of sample microcell geometric position and spectral information is detected, and reach resolving power and measurement model simultaneously That encloses effectively takes into account.But the test method makes full use of the effective numerical aperture for measuring object lens due to failing, and affects confocal The spatial resolution of Raman spectrum system;In addition, the test macro is required strictly by off-axis position, the factors such as adjustment difficulty Limitation.
Summary of the invention
It is larger the purpose of the present invention is being limited for the solution prior art by extraneous factor, and the problem that spatial resolution is low, It is proposed postposition light splitting pupil laser differential confocal Raman spectra test method and device.By being lost in confocal Raman spectra detection system The Rayleigh scattering light building light splitting pupil differential confocal microscopic imaging system of abandoning, realizes that the high-space resolution of sample geometrical morphology is visited It surveys;Sample focal point is accurately obtained using the focal position that postposition light splitting pupil laser differential confocal micro imaging system obtains Raman spectral information, and then realize light splitting pupil differential confocal Raman spectra high-space resolution imaging and the detection of " collection of illustrative plates ".
The purpose of the present invention is what is be achieved through the following technical solutions.
Postposition is divided pupil laser differential confocal Raman spectra test method, comprising the following steps:
Pupil is collected Step 1: placing on collecting object lens pupil plane first, the excitation beam of light-source system outgoing passes through Amici prism and measurement object lens after, be focused on sample, inspire be loaded with sample microcell characterisitic parameter information Raman dissipate Light is penetrated, while reflecting Rayleigh scattering light;Raman diffused light and Rayleigh scattering light are collected by measurement object lens, by Amici prism It after reflection, is divided into two bundles by dichroic optical system, wherein after the Rayleigh scattering light of dichroic optical system reflection enters Light splitting pupil laser differential confocal detection system is set, after collecting object lens and collecting pupil, is carried out by intensity collection system burnt Spot segmentation detection, realizes the detection to sample microcell geometric position.Detailed process are as follows: intensity collection system acquisition is gathered Burnt hot spot is split processing, respectively obtains search coverage A light intensity signal corresponding with search coverage B, to two detecting areas of A, B The signal progress in domain is differential to subtract each other processing, obtains light splitting pupil differential confocal curve I (z), and wherein z is axial physical coordinates.It utilizes " zero crossing " for being divided pupil differential confocal curve accurately corresponds to characteristic with measurement object focal point, is triggered by " zero crossing " come accurate Capture excitation hot spot focal position, and then realize the geometry detection and space orientation of high-space resolution.
At the same time, the light beam of the Raman diffused light by dichroic optical system transmission enters Raman spectroscopic detection system In system.Postposition light splitting pupil laser differential confocal detection system accurately captures the spectrum of focal beam spot by the position of " zero crossing " Information, and then realize the spectrographic detection to sample microcell high spatial.Detailed process are as follows: by carrying out axial scan acquisition to sample Postposition is divided the axial response curve of pupil laser differential confocal detection system, is carried out by " zero crossing " of curve to sample Real-time tracking fixed-focus.The spatial position of sample is controlled, by 3 D scanning system to guarantee to begin in entire measurement process The Raman spectral information at focal position is obtained eventually, and then inhibits system drifting and the external world caused by long-time spectral measurement The influence of environment improves the measurement accuracy and spatial resolution of system.
At the laser facula only obtained to the postposition light splitting pupil laser differential confocal detection system for receiving Reyleith scanttering light When reason, it is able to carry out the chromatography detection of high-space resolution three-dimensional;
When the raman spectral signal that the Raman spectroscopic detection system that only Raman diffused light is received in docking obtains is handled, energy It is enough that spectrographic detection is carried out to sample;
When simultaneously to receive Reyleith scanttering light postposition light splitting pupil laser differential confocal detection system obtain light intensity signal with connect When the spectral signal that the Raman spectroscopic detection system of receipts Raman diffused light obtains is handled, it is able to carry out high-space resolution microcell Map tomography realizes that the postposition for carrying out " collection of illustrative plates " to sample is divided pupil laser differential confocal Raman spectrum high-altitude Between resolution imaging and detection.
Particularly, in the methods of the invention, collecting pupil can be round, D-shaped or other shapes.
In the methods of the invention, excitation beam includes light beams such as line polarisation, rotatory polarization, radial polarisation light and by pupil Thus the structure light beam that the technologies such as filtering generate improves system spectrum Signal-to-Noise and system transverse resolution.
In the methods of the invention, can also detect including fluorescence, Brillouin scattering, Compton scattering light scattering light Spectrum.
In the methods of the invention, that the object lens of the measurement to different NA can be realized only is handled by computer system software Match, without carrying out any hardware adjustment to system.
The present invention provides postpositions to be divided pupil laser differential confocal Raman spectroscopy test device, including generating excitation beam Light-source system, measurement object lens, 3 D scanning system, dichroic optical system, Raman spectroscopic detection system, postposition are divided pupil laser Differential confocal measurement system, computer processing system.
Wherein, it collects pupil to be placed on the pupil plane for collecting object lens, sample is fixed on the load of 3 D scanning system On object platform.Dichroic optical system is placed on the light beam reflection direction of Amici prism, and Raman spectroscopic detection system is placed on two To in the transmission direction of color beam splitting system, postposition light splitting pupil laser differential confocal measuring system is placed on dichroic optical system On reflection direction, computer processing system is used to swash 3 D scanning system, Raman spectroscopic detection system and postposition light splitting pupil The equation of light moves confocal measuring system and carries out data processing and coordinated control.
In apparatus of the present invention, intensity collection system can use method of the pin hole in conjunction with light intensity point detector, realize Segmentation detection to Airy.
In apparatus of the present invention, intensity collection system can use ccd detector, be detected by being arranged in CCD detection face The position of region A and search coverage B and size realize that the segmentation to Airy detects.
In apparatus of the present invention, intensity collection system can using conduction optical fiber, by the focal plane of third convergent mirror, About the symmetrically placed first conduction optical fiber of optical axis and the second conduction optical fiber, realize that the segmentation to Airy detects.
In apparatus of the present invention, magnifying glass can be relayed by increasing, the Aili spot that amplification intensity collection system detects, with Improve the acquisition precision of postposition light splitting pupil laser differential confocal measuring system.
Beneficial effect
1, the present invention has merged postposition light splitting pupil laser differential confocal microtechnic and Raman spectroscopic detection technology, by rear " zero crossing " for setting light splitting pupil laser differential confocal axial response curve accurately corresponds to this with the focus of high-acruracy survey object lens Characteristic realizes accurate fixed-focus to sample, while obtaining geometric position and the raman spectral signal of sample, existing greatly improving While having the microscopic spectrum detectivity of confocal Raman spectra detection system, the light channel structure of system is also greatly simplified, this It is one of the innovative point for being different from existing confocal Raman spectra Detection Techniques;
2, the present invention is divided pupil laser differential confocal measuring system by the way that postposition is added in collecting optical path, in conventional confocal It realizes that postposition is divided pupil laser differential confocal micro-imaging on the basis of Raman system, it is poor to further increase postposition light splitting pupil laser The spatial resolution of dynamic crs technique, this be different from existing confocal Raman spectra Detection Techniques innovative point it Two;
3, the present invention obtains signal using the method for division focal spot, by hot spot focal plane in setting intensity collection system The size of tiny area extends its application field to match different sample reflectivity;At computer system software The matching of the measurement object lens to different NA values may be implemented in reason, without carrying out any hardware adjustment, Jin Erti to system again The versatility of high instrument.This be different from existing confocal Raman spectra Detection Techniques innovative point three;
4, the present invention by postposition be divided pupil laser differential confocal microscopic system and Raman spectroscopic detection system in structure and Functionally combine, it can be achieved that the micro- detection of sample microcell postposition light splitting pupil laser differential confocal, confocal laser Raman spectroscopic detection, Postposition is divided the switching of pupil laser differential confocal Raman spectroscopic detection various modes, this is to be different from existing confocal Raman spectra detection The four of the innovative point of technology.
Detailed description of the invention
Fig. 1 is conventional confocal Raman spectrum imaging method schematic diagram;
Fig. 2 is that existing postposition is divided pupil laser differential confocal Raman spectra test method schematic diagram;
Fig. 3 is that postposition of the present invention is divided pupil laser differential confocal Raman spectra test method schematic diagram, that is, is made a summary Attached drawing;
Fig. 4 is that D-shaped postposition is divided pupil laser differential confocal Raman spectra test method schematic diagram;
Fig. 5 is the postposition light splitting pupil laser differential confocal Raman spectra test method schematic diagram in conjunction with iris filter;
Fig. 6 is that the postposition with beam modulation system is divided pupil laser differential confocal Raman spectroscopy test device schematic diagram;
Fig. 7 is that the postposition with detection focal spot amplification system is divided the signal of pupil laser differential confocal Raman spectroscopy test device Figure;
Fig. 8 is to be divided pupil laser differential confocal Raman spectroscopy test device using the postposition of pin hole and light intensity point detector to show It is intended to;
Fig. 9 is to be divided pupil laser differential confocal Raman spectroscopy test device schematic diagram using the postposition of ccd detector;
Figure 10 is that the postposition detected using optical fiber is divided pupil laser differential confocal Raman spectroscopy test device schematic diagram;
Figure 11 is that high-space resolution postposition is divided pupil laser differential confocal Raman spectra test method and schematic device, i.e., Embodiment figure;
Figure 12 is light splitting pupil differential confocal simulation curve;
Figure 13 is light splitting pupil differential confocal measured curve.
Wherein, 1- light-source system, 2- Amici prism, 3- measure object lens, 4- sample, 5- 3 D scanning system, 6- bis- To color beam splitting system, 7- Raman spectroscopic detection system, the first convergent mirror of 8-, the first pin hole of 9-, the second convergent mirror of 10-, 11- spectrum Probe unit, 12- postposition light splitting pupil laser differential confocal detection system, 13- collects object lens, 14- collects pupil, 15- light intensity is adopted Burnt curve, 20- are divided after burnt curve, 19- light splitting pupil before collecting system, 16- search coverage A, 17- search coverage B, 18- light splitting pupil Pupil differential confocal curve, 21- radial polarisation converting system, 22- iris filter, 23- beam modulation system, 24- third are assembled Mirror, the second pin hole of 25-, the 4th convergent mirror of 26-, 27- relay magnifying glass, 28- third pin hole, 29- light intensity point detector, 30-CCD Detector, 31- first conduct optical fiber, 32- second conducts optical fiber, 33- computer, 34- quarter-wave plate, 35- assemble object lens, 36- illumination iris, 37- detect pupil, the 5th convergent mirror of 38-, 39- light intensity detection system, 40- laser spot detection region A, 41 hot spots The off-axis confocal axial curve of search coverage B, 42- first, 43 second off-axis confocal axial curves, 44 differential confocal axial characteristics are rung Burnt measured curve, 47- are divided pupil differential confocal measured curve after burnt measured curve, 46- light splitting pupil before answering curve, 45- to be divided pupil.
Specific embodiment
Invention is further described in detail with reference to the accompanying drawings and examples.
Pupil 14 is collected as shown in figure 3, placing on collecting 13 pupil plane of object lens.The excitation beam that light-source system 1 is emitted It after through Amici prism 2 and measurement object lens 3, is focused on sample 4, inspires and be loaded with sample microcell characterisitic parameter letter The Raman diffused light of breath, while reflecting Rayleigh scattering light.Raman diffused light and Rayleigh scattering light are collected by measurement object lens, warp It after crossing the reflection of Amici prism 2, is divided into two bundles by dichroic optical system 6, wherein the Rayleigh reflected by dichroic optical system 6 Scattering light enters postposition light splitting pupil laser differential confocal detection system 12, after collecting object lens 13 and collecting pupil 14, quilt Intensity collection system 15 carries out focal spot segmentation detection, realizes the detection to sample microcell geometric position.By dichroic point The light beam for the Raman diffused light that photosystem 6 transmits enters in Raman spectroscopic detection system 7, is divided pupil laser differential using postposition " zero crossing " for the light splitting pupil differential confocal curve 20 that confocal detection system 12 obtains accurately corresponding object lens focal beam spot focus this One characteristic is accurately captured the spectral information of focal beam spot by the position of " zero crossing ", and then realized to sample microcell high-altitude Between spectrographic detection.
Particularly, it can will collect pupil 14 and replace other shapes, such as D-shaped, is i.e. composition D-shaped postposition is divided pupil laser differential Confocal Raman spectra test method, as shown in Figure 4.
Particularly, by addition radial polarisation converting system 21 and iris filter 22, to further increase spectrographic detection Spatial resolution, that is, constitute addition iris filter postposition be divided pupil laser differential confocal Raman spectra test method, such as Shown in Fig. 5.
Particularly, have by adding beam modulation system 23 between light-source system 1 and Amici prism 2, and then constituting The postposition of beam modulation system is divided pupil laser differential confocal Raman spectroscopy test device.As shown in fig. 6, beam modulation system 23 Including third convergent mirror 24, the second pin hole 25, the 4th convergent mirror 26 being sequentially placed along optical path.
Particularly, increase relaying magnifying glass 27 in postposition light splitting pupil laser differential confocal detection system 12, after improving The acquisition precision of light splitting pupil laser differential confocal detection system is set, as shown in Figure 7.
Particularly, the method combined using pin hole 28 with light intensity point detector 29 realizes that the segmentation to Airy detects, such as Shown in Fig. 8.
Particularly, intensity collection system 15 can use ccd detector 30, set micro- on detection focal plane by changing The parameter of zonule is realized that the segmentation to Airy detects, is answered so as to extend it to match the reflectivity of different samples With field, as shown in Figure 9.
Particularly, intensity collection system 15 can using conduction optical fiber, by collect object lens 13 focal plane at, about light Axial symmetry places the first conduction optical fiber 31 and the second conduction optical fiber 32 realizes that the segmentation to Airy detects, as shown in Figure 10.
Embodiment
In the present embodiment, light-source system 1 uses 532nm continuous wave laser, and dichroic optical system 6 uses Notch Filter, spectrographic detection unit 11 use Raman spectrometer.
As shown in figure 11, postposition light splitting pupil laser differential confocal Raman spectroscopic detection process is as follows:
Firstly, the laser that the light-source system 1 being made of laser is emitted is assembled by third convergent mirror 24 and enters the second needle Behind hole 25, parallel excitation beam is formed by 26 collimator and extender of the 4th convergent mirror.Excitation beam is by radial polarisation conversion system After system 21, Amici prism 2 and iris filter 22, be measured object lens 3 and be focused on sample 4, inspire be loaded with it is tested The Raman diffused light of 4 microcell characterisitic parameter of sample.
Then the mobile sample 4 of 3 D scanning system 5 is controlled by computer 33, makes different zones on sample 4 Raman diffused light and the Reyleith scanttering light in the corresponding region be measured object lens 3 and collect, by iris filter 22 and light splitting rib After mirror 2 reflects, Reyleith scanttering light is carried out lossless separate with Raman diffused light by Notch Filter6.
Wherein the Reyleith scanttering light by Notch Filter6 reflection enters postposition light splitting pupil laser differential confocal detection system 12 In, relaying magnifying glass 27 is entered after collecting object lens 13 and collecting pupil 14, the hot spot being amplified is connect by ccd detector 30 It receives, passes through the x in detection focal spotdAbout y on axisdTwo small search coverage A16 and search coverage B17 are arranged in axial symmetry, It is relative to ydAxis deviates vM, the Intensity response of two search coverages is respectively IA(u,-vM) and IB(u,+vM), computer 33 It is differential to obtained light intensity signal progress to subtract each other, obtain postposition light splitting pupil laser differential confocal curve 20:
I(u,vM)=IA(u,-vM)-IB(u,+vM)
Wherein, I (u, vM) it is that postposition is divided pupil laser differential confocal curve, u is axial normalized coordinate, vMFor pin hole cross To normalized offset.In conjunction with the location information of the 3 D scanning system 5 obtained by computer 33, obtains postposition light splitting pupil and swash The equation of light moves the differential confocal curve 20 of confocal detection system and its position of " zero crossing ";It is divided pupil differential confocal curve 20 " zero crossing " and the focal beam spot of excitation beam are accurately corresponding, thus by " zero crossing " of differential confocal curve 20 can be obtained by The elevation information of sample 4, and then reconstruct the three-dimensional appearance of sample 4.Figure 13 is light splitting pupil differential confocal measured curve figure, Wherein 45 be that burnt measured curve, 46 are that burnt measured curve, 47 are to be divided pupil differential confocal measured curve after being divided pupil before being divided pupil.
Raman diffused light by Notch Filter6 transmission passes through the first convergent mirror 8, the first pin hole 9 and the second meeting It is received after poly- mirror 10 by Raman spectrometer 11, measures the raman scattering spectrum for being loaded with 4 microscopic spectrum characteristic of sample.
At the laser facula only obtained to the postposition light splitting pupil laser differential confocal detection system 12 for receiving Reyleith scanttering light When reason, the chromatography detection of high-space resolution three-dimensional can be carried out to sample 4;When only the Raman spectrum of Raman diffused light is received in docking When the raman spectral signal that instrument 11 obtains is handled, spectrographic detection can be carried out to sample 4;Simultaneously to reception Reyleith scanttering light Postposition light splitting pupil laser differential confocal detection system 12 obtain light intensity signal and receive Raman diffused light Raman spectrometer When 11 spectral signals obtained are handled, high-space resolution microcell map tomography can be carried out to sample 4, in turn Postposition light splitting pupil laser differential confocal Raman spectrum high-space resolution imaging and the detection of " collection of illustrative plates " are carried out to sample.
As shown in figure 11, postposition light splitting pupil laser differential confocal Raman spectroscopy test device includes the light for generating excitation beam Source system 1, beam modulation system 23, radial polarisation converting system 21, Amici prism 2, iris filter 22, measurement object lens 3, quilt Sample 4,3 D scanning system 5, Notch Filter6, postposition are divided pupil laser differential confocal detection system 12, Raman spectrum Detection system 7.
More than, along the laser emitting direction of light-source system 1, it is sequentially placed third convergent mirror 24, the second pin hole 25, the 4th meeting Poly- mirror 26, radial polarisation converting system 21, Amici prism 2, iris filter 22, measurement object lens 3, sample 4 and three-dimensional Scanning system 5 is sequentially placed Notch Filter6, the first convergent mirror 8, the first pin hole 9, second in 2 reflection direction of Amici prism Convergent mirror 10, Raman spectrometer 11, Notch Filter6 reflection direction be sequentially placed collect object lens 13, collect pupil 14, in After magnifying glass 27, ccd detector 30, wherein collecting the optical axis of pupil 14 and the optical axis coincidence of Notch Filter6 the reflected beams; In the entire system, continuous wave laser 1,3-D scanning translation stage 5, Raman spectrometer 11 and ccd detector 30 are calculated Machine 33 controls, and the three dimensional local information and spectral information that system obtains also carry out fusion treatment by computer 33.
A specific embodiment of the invention is described in conjunction with attached drawing above, but these explanations cannot be understood to limit The scope of the present invention, protection scope of the present invention are 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 (9)

1. postposition is divided pupil laser differential confocal Raman spectra test method, it is characterised in that: the following steps are included:
Pupil (14) are collected Step 1: placing on collecting object lens (13) pupil plane;The excitation beam warp of light-source system (1) outgoing It crosses by being focused on sample (4) behind Amici prism (2) and measurement object lens (3), inspires that be loaded with sample (4) micro- The Raman diffused light of zone properties parameter information, while reflecting Rayleigh scattering light;Raman diffused light and Rayleigh scattering light are by surveying Measure object lens (3) collect, by Amici prism (2) reflection after, be divided into two bundles by dichroic optical system (6), wherein by two to The Rayleigh scattering light of color beam splitting system (6) reflection enters light splitting pupil laser differential confocal detection system (12), by collecting object lens (13) and after collection pupil (14), focal spot segmentation detection is carried out by intensity collection system (15), is realized micro- to sample (4) The detection of area geometric position;
To the method for the detection of sample (4) microcell geometric position described in step 1 are as follows:
The focal beam spot obtained to intensity collection system (15) is split processing, respectively obtains search coverage A (16) and detection The corresponding light intensity signal of region B (17), it is differential to two signals progress to subtract each other processing, obtain light splitting pupil differential confocal curve (20);Characteristic is accurately corresponded to measurement object lens (3) focus using " zero crossing " of light splitting pupil differential confocal curve (20), is passed through " zero crossing " triggering accurately to capture excitation hot spot focal position, and then realizes that the geometry detection of high-space resolution and space are fixed Position;
Step 2: at the same time, the light beam of the Raman diffused light by dichroic optical system (6) transmission enters Raman spectrum In detection system (7);The light splitting pupil differential confocal curve (20) obtained using light splitting pupil laser differential confocal detection system (12) " zero crossing " accurate corresponding measurement this characteristic of object lens (3) focal beam spot focus, accurately captured by the position of " zero crossing " The spectral information of focal beam spot, and then realize the spectrographic detection to sample (4) microcell high spatial;
To the method for the spectrographic detection of sample (4) microcell high spatial described in step 2 are as follows: by carrying out axial sweep to sample The differential confocal curve (20) for obtaining light splitting pupil laser differential confocal detection system is retouched, by " zero crossing " of curve to detected sample Product carry out real-time tracking fixed-focus;By the spatial position of 3 D scanning system (5) control sample (4), to guarantee entire Always the Raman spectral information at focal position is obtained in measurement process, and then inhibits system caused by long-time spectral measurement Drift and the influence of external environment, improve the measurement accuracy and spatial resolution of system;
When the laser facula only obtained to the light splitting pupil laser differential confocal detection system (12) for receiving Reyleith scanttering light is handled, Sample (4) can be carried out with the chromatography detection of high-space resolution three-dimensional;
When the raman spectral signal that the Raman spectroscopic detection system (7) that only Raman diffused light is received in docking obtains is handled, energy It is enough that spectrographic detection is carried out to sample (4);
The light intensity signal that the light splitting pupil laser differential confocal detection system (12) for receiving Reyleith scanttering light obtains is drawn with reception when simultaneously When the spectral signal that the Raman spectroscopic detection system (7) of graceful scattering light obtains is handled, it is able to carry out high-space resolution microcell Map tomography, and then sample (4) are carried out with the light splitting pupil laser differential confocal Raman spectrum high-altitude of " collection of illustrative plates " Between resolution imaging and detection.
2. a kind of light splitting pupil differential laser differential confocal Raman spectra test method according to claim 1, feature exist In: the system can also detect scattering spectrum.
3. a kind of light splitting pupil differential laser differential confocal Raman spectra test method according to claim 1, feature exist In: the shape for collecting pupil (14) includes round or D-shaped.
4. a kind of light splitting pupil differential laser differential confocal Raman spectra test method according to claim 1, feature exist In: the excitation beam includes light beam and structure light beam, thus improves system spectrum Signal-to-Noise and system is laterally divided Distinguish power.
5. postposition is divided pupil laser differential confocal Raman spectroscopy test device, it is characterised in that: the light including generating excitation beam Source system (1), Amici prism (2), measurement object lens (3), sample (4), 3 D scanning system (5), dichroic optical system (6), pupil laser differential confocal detection system (12), Raman spectroscopic detection system (7) and computer processing system (33) are divided;
The light splitting pupil laser differential confocal detection system (12) includes collecting object lens (13), collecting pupil (14) and intensity collection System (15);
The Raman spectroscopic detection system (7) includes the first convergent mirror (8), the first pin hole (9), the second convergent mirror (10) and spectrum Probe unit (11);
Sample (4) is placed on 3 D scanning system (5);The excitation beam of light-source system (1) outgoing passes through through light splitting rib After mirror (2) and measurement object lens (3), it is focused on sample (4), inspires the Raman for being loaded with sample microcell characterisitic parameter information Light is scattered, while reflecting Rayleigh scattering light;Raman diffused light and Rayleigh scattering light are collected by measurement object lens, by being divided rib It after mirror (2) reflection, is divided into two bundles by dichroic optical system (6), wherein the Rayleigh by dichroic optical system (6) reflection dissipates It penetrates light and enters light splitting pupil laser differential confocal detection system (12);The Raman diffused light transmitted by dichroic optical system (6) Light beam enter in Raman spectroscopic detection system (7).
6. postposition according to claim 5 is divided pupil laser differential confocal Raman spectroscopy test device, it is characterised in that: also Including radial polarisation converting system (21) and iris filter (22);Radial polarisation converting system (21) is placed in light-source system (1) Between Amici prism (2);The iris filter (22) is placed between Amici prism (2) and measurement object lens (3).
7. postposition according to claim 5 is divided pupil laser differential confocal Raman spectroscopy test device, it is characterised in that: also Including beam modulation system (23);The beam modulation system (23) is placed between light-source system (1) and Amici prism (2);Institute Beam modulation system (23) is stated to be made of third convergent mirror (24), the second pin hole (25) and the 4th convergent mirror (26).
8. postposition according to claim 5 is divided pupil laser differential confocal Raman spectroscopy test device, it is characterised in that: light Strong acquisition system (15) can be by ccd detector (30), the first conduction optical fiber (31) and the second conduction optical fiber (32) or third needle Hole (28) and light intensity point detector (29) are replaced.
9. postposition according to claim 5 is divided pupil laser differential confocal Raman spectroscopy test device, it is characterised in that: institute Stating light splitting pupil laser differential confocal detection system (12) further includes relaying magnifying glass (27), and relaying magnifying glass (27) is placed in gleanings Between mirror (13) and intensity collection system (15).
CN201811343472.1A 2018-11-13 2018-11-13 Postposition is divided pupil laser differential confocal Raman spectra test method and device Pending CN109211873A (en)

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