CN106442467A - Spatial autofocusing laser confocal imaging Raman-spectrum detecting method and device - Google Patents

Spatial autofocusing laser confocal imaging Raman-spectrum detecting method and device Download PDF

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CN106442467A
CN106442467A CN201611023198.0A CN201611023198A CN106442467A CN 106442467 A CN106442467 A CN 106442467A CN 201611023198 A CN201611023198 A CN 201611023198A CN 106442467 A CN106442467 A CN 106442467A
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raman
confocal
sample
autofocusing
light
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CN106442467B (en
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祝连庆
王帅
姚齐峰
张雯
董明利
娄小平
骆飞
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Beijing Information Science and Technology University
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Beijing Information Science and Technology University
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    • 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
    • G01N21/62Systems 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

Abstract

The invention discloses a spatial autofocusing laser confocal imaging Raman-spectrum detecting method and a spatial autofocusing laser confocal imaging Raman-spectrum detecting device. According to the method and the device, through introducing a focusing telescopic technique and a confocal technique into spectral detection, separation is carried out on Rayleigh scattered light and Raman scattered light by utilizing a dichroic light splitting system; the automatic focus adjustment of a telescopic focusing system is precisely controlled by utilizing the characteristic that the maximum value of a confocal response curve of a detector precisely corresponds to a focal position and through searching and responding to the maximum value; exciting light beams are automatically focused on a to-be-detected object; meanwhile, spectral information of the focal position of a light spot of laser is obtained; meanwhile, through the light splitting system and through an imaging system and an image sensing system, image collection in the spatial area of the sample is obtained; the spatial autofocusing spectral detection and image obtaining are realized; a method and a device which are capable of realizing the spatial autofocusing imaging spectral detection of the sample are formed. The method and the device have the characteristics of automatic focusing and accurate positioning; the detection range is enlarged; the sensitivity of the spectral detection is improved.

Description

Space autofocusing confocal laser image Raman optical spectrum detecting method and device
Technical field
The present invention relates to space optics imaging and spectral measurement methodses field are and in particular to a kind of space autofocusing laser is common Burnt image Raman optical spectrum detecting method and device.
Background technology
Aerial image technology is combined by confocal laser Raman spectrum measuring technology with Raman spectrum analyses technology New technique, incident laser is focused on sample by autofocusing focusing system of looking in the distance by it, such that it is able in larger distance In the case of not being subject to ambient substance interference, obtain material composition structure and composition of product etc. in the same old way, preferable molecule is provided " fingerprint " feature.It not only can in the same aspect of observing samples different microcells raman spectral signal moreover it is possible to observe sample respectively The Raman signal of the different aspect of product spatial depth, carries out spacescan to sample, thus the situation in not lesioned sample It is issued to the effect carrying out collection of illustrative plates detection.Confocal laser Raman spectrum measuring technology due to its lossless spectrum tomography ability and High-resolution, be widely used to physics, chemistry, biomedicine, petrochemical industry, environmental science, material science, geology, Xing detect, The field such as archaeology and gemstone testing.
Existing confocal laser Raman spectrometer employs microscopic system at present, limits system detectable range;Using three Dimension mobile platform, as sample carrying platform, limits sample size and existing forms;It is fixed to be carried out using weak Raman diffused light Position, reduce system focuses sensitivity;;During long-time spectrographic detection, system easily is affected to occur by factors such as environment Drift, produces out of focus, reduces the reliability of system long-term work;System only carries out spectrographic detection, and pattern is single;Measurement process In need to hide light, working environment is restricted.
Content of the invention
The present invention provides a kind of space self-regulated confocal laser image Raman optical spectrum detecting method with device it is therefore intended that solving The problem that existing confocal Raman spectra Detection Techniques investigative range is difficult to improve and spectrographic detection pattern is single.
The technical scheme is that:A kind of space autofocusing confocal laser image Raman spectral detection device includes exciting Beam system, focusing system of looking in the distance, dichroic optical system, the first beam splitting system, the second beam splitting system, Raman spectroscopic detection system System, confocal detection system and image sensing and data processing module;Wherein, excitation beam system and focusing system edge of looking in the distance Light path is placed sequentially in the reflection direction of dichroic optical system;First beam splitting system is in the transmission side of dichroic optical system To;Raman spectroscopic detection system is located at the transmission direction of the first beam splitting system;Second beam splitting system is located at the first beam splitting system Reflection direction;Confocal detection system is located at the transmission direction of the second beam splitting system;Image sensing is located at the second beam splitting system Reflection direction;Data processing module and Raman spectroscopic detection system, image sensing, confocal detection system and focusing of looking in the distance System connects.
Preferably, described excitation beam system also includes light polarization modulator and iris filter.For producing polarized light And space structure light beam, for improving the optical property of system.
Preferably, the described iris filter exciting hot spot for compression is located at Polarization Controller and dichroic light splitting system Between system or be located at dichroic optical system and look in the distance between focusing system.
Preferably, excitation beam system and focusing system of looking in the distance also are placed in the transmission of dichroic optical system successively along light path Direction, the first beam splitting system is placed sequentially in the reflection direction of dichroic optical system, and Raman spectroscopic detection system is located at first The transmission direction of beam splitting system, the second beam splitting system is located at the reflection direction that confocal detection system is located at the first beam splitting system, altogether Burnt detection system is located at the transmission direction of the second beam splitting system, and image sensing is located at the reflection direction of the second beam splitting system.
Preferably, Raman spectroscopic detection system is common Raman spectroscopic detection system, including be sequentially placed along light path 5th condenser lenss, the first spectrogrph positioned at the 5th condenser lenss focal position and the second detector after the first spectrogrph, Detection for the surface spectrum of sample;Or confocal Raman spectra detection system, including being sequentially placed along light path Seven condenser lenss, positioned at the 3rd pin hole of the 7th condenser lenss focal position, the 8th condenser lenss after the 3rd pin hole, positioned at the 8th Second spectrogrph of condenser lenss focal position and be located at the second spectrogrph after the 3rd detector, for improve system signal noise ratio and Spatial resolution, completes the spectrographic detection to sample.
Preferably, data processing module includes for the confocal data processing module of processing position information and is used for processing position Confidence breath and spectral information data fusion module, also include for control look in the distance focusing system focusing data control block and Image module.
A kind of space autofocusing confocal laser image Raman optical spectrum detecting method, the light using focusing system of looking in the distance collects energy Power;Dichroic optical system by systematic collection to scattered light be separated into Rayleigh scattering light and Raman diffused light;Described Rayleigh dissipates Penetrate light entrance confocal detection system and carry out the adjustment of telescope focal position and the focusing of exciting light;Described Raman diffused light enters Raman spectroscopic detection system carries out spectrographic detection;Image sensing obtains sample space image, using confocal curves maximum M With focus O position accurately this characteristic corresponding, focus on sample by maximizing come precise control excitation beam, simultaneously Obtain the spectral information exciting hot spot focal position, and complete sample space image acquisition using image sensing;Described side Method comprises the steps:
1) excitation beam is produced by excitation beam system, after dichroic optical system and focusing system of looking in the distance, shine Penetrate on sample, and inspire Rayleigh scattering light and the Raman diffused light being loaded with sample spectra characteristic;
2) Rayleigh scattering light in corresponding sample region and Raman diffused light is made to again pass by focusing system of looking in the distance, and quilt Focusing system of looking in the distance is shaped to directional light and is transmitted through dichroic optical system, and draws to Rayleigh scattering light through dichroic optical system Graceful scattered light carries out separating;
3) part Rayleigh scattering light, by dichroic optical system transmission, reflects through the first beam splitting system, through the second light splitting system System is transmitted into confocal detection system, using the first detector in confocal detection system, records reflection sample position information Intensity response I [α, β, l];
4) pass through focus adjusting mechanism, make the response of confocal detection system maximum, complete excitation beam and be autofocusing on sample, Obtain the positional information [α, β, l] of sample simultaneously;
5) complete after excitation beam focuses, image sensing obtain sample space image information P (α, β, l);
6) Raman diffused light is transmitted into Raman spectroscopic detection through dichroic optical system transmission, then through the first beam splitting system System, records the Raman scattering signal I (λ) being loaded with sample characteristic, you can carry out spectrum using Raman spectroscopic detection system Test, wherein λ is wavelength;
7) I (λ) is sent to data processing module and carries out data processing, thus obtain comprising sample corresponding region position The spectral information I (λ) putting, object location information [α, β, l];
8) sample is carried out along α, β scanning direction, focusing system of looking in the distance carries out focusing along l scanning direction, repeat on State the one group of n sequence measuring information [I comprising positional information [α, β, l] and I (λ) that step records corresponding objective focus positions (λ), α, β, l] and Pn (α, β, l);
9) utilize the corresponding positional information [α, β, l] of distinguishable region δ n, find out the spectral information In (λ) in corresponding δ n region Value, further according to the relation with space coordinatess [α, β, l], the information of reconstruct reflection measured object microcell δ n three dimensional structure and spectral characteristic In (α n, β n, ln, λ n), that is, achieve spectrographic detection and the three-dimensional geometry position sensing of microcell δ min;
10) three dimension scale of corresponding minimum distinguishable region δ min and spectral characteristic are determined by following formula:
Achieve large space scope confocal imaging Raman spectroscopic detection.
Preferably, confocal curves maximum value position correspondence is looked in the distance focusing system focal position, and focused spot size is the most herein Little, the region of detection is minimum, and confocal curves other positions correspondence is looked in the distance the out of focus region of focusing system, before Jiao or defocused region Interior focused spot size increases with defocusing amount and increases.Using this feature, by adjusting the focus adjusting mechanism of focusing system of looking in the distance, Accurately excitation beam is focused on sample.
Preferably, described excitation beam is polarized beam:Linear polarization, circular polarization or radial polarisation light, or by pupil The structure light beam that filtering technique generates, it can compress the size of measurement focal beam spot with polarization Modulation combination, improves system Unite angular resolving power.
The invention has the beneficial effects as follows:A kind of space autofocusing confocal laser image Raman optical spectrum detecting method and device, Merge technology, focus technique, confocal technology and spectrographic detection technology and the image sensing technology of looking in the distance;Using looking in the distance, focusing system carries High system light capacity gauge, makes system have large space investigative range;Using image sensing technology, obtain sample space image letter Breath;Using being accurately positioned of confocal system focusing, the spatial resolution of spectrographic detection is greatly improved;The confocal skill of system globe area Art, focus technique, can achieve and automatically focus on, realize the Auto-focusing detecting of sample;System aerial image simultaneously, collection of illustrative plates imaging and Spectrum test Three models.
Brief description
With reference to the accompanying drawing enclosed, the more purpose of the present invention, function and advantage will be as follows by embodiment of the present invention Description is illustrated, wherein:
Fig. 1 illustrates confocal laser response curve of the present invention;
Fig. 2 illustrates space of the present invention autofocusing confocal laser image Raman optical spectrum detecting method schematic diagram;
Fig. 3 illustrates space of the present invention autofocusing confocal laser image Raman spectral detection device schematic diagram;
Fig. 4 illustrates that space of the present invention autofocusing confocal laser image Raman optical spectrum detecting method is illustrated with device embodiment 1 Figure;
Fig. 5 illustrates that space of the present invention autofocusing confocal laser image Raman optical spectrum detecting method is illustrated with device embodiment 2 Figure.
Specific embodiment
By reference to one exemplary embodiment, the purpose of the present invention and function and the side for realizing these purposes and function Method will be illustrated.However, the present invention is not limited to one exemplary embodiment disclosed below;Can by multi-form Lai It is realized.The essence of description is only to aid in the detail of the various equivalent modifications Integrated Understanding present invention.
Hereinafter, embodiments of the invention will be described with reference to the drawings.In the accompanying drawings, identical reference represent identical Or similar part, or same or similar step.
The present invention is described further with reference to the accompanying drawings and examples.
Fig. 1 is confocal laser response curve of the present invention, the focusing as shown in figure 1, confocal curves maximum value position M correspondence is looked in the distance System focus position O.
Fig. 2 is space of the present invention autofocusing confocal laser image Raman optical spectrum detecting method schematic diagram.As shown in Fig. 2 swashing Luminous beam system 600 produces exciting light, reflects through dichroic optical system 900, after focusing system 100 of looking in the distance, focuses on On sample 140, and Rayleigh scattering light and the Raman diffused light being loaded with sample spectral characteristic are inspired on sample, swash The Raman diffused light sending and Rayleigh scattering light, by systematic collection recovering light path, divide with dichroic after focusing system 100 of looking in the distance After photosystem 900 transmission, Raman diffused light and part Rayleigh scattering light are through the first beam splitting system 150 light splitting, part Rayleigh scattering Light is reflected into the second beam splitting system 160, and wherein fractional transmission entrance confocal detection system 170 carries out position sensing, part Image sensing 310 is reflected into by the second beam splitting system 160, Raman diffused light is transmitted into spectrum investigating system 220 and enters Row spectrographic detection, according to confocal response curve, data processing module 340 controls focus adjusting mechanism 120 of looking in the distance to focus, and so that exciting light is gathered Jiao, on sample, makes confocal response curve maximum, completes the automatic focusing of excitation beam.
Fig. 3 is space of the present invention autofocusing confocal laser image Raman spectral detection device schematic diagram.As shown in figure 3, this Device includes the excitation beam system 600 being sequentially placed along light path, dichroic optical system 900, focusing system 100 of looking in the distance, quilt Test sample product 140, positioned at the first beam splitting system 150 of dichroic optical system 900 transmission direction, positioned at the first beam splitting system 150 The spectrum investigating system 220 of transmission direction and the second beam splitting system 160 of reflection direction, positioned at the second beam splitting system 160 transmission The confocal detection system 170 in direction, positioned at the image sensing 310 of the second beam splitting system 160 reflection direction, also includes connecting Spectrum investigating system 220, confocal detection system 170, image sensing 310, the data processing module of focusing system 100 of looking in the distance 340 and computer control system 350.
Embodiment 1
In the present embodiment, dichroic optical system 900 is optical notch filter, and the first beam splitting system 150 is to have necessarily The broadband beam splitting system of splitting ratio, the second beam splitting system 160 is the broadband beam splitting system with certain splitting ratio.
Fig. 4 is space of the present invention autofocusing confocal laser image Raman optical spectrum detecting method and device embodiment 1 schematic diagram.
As shown in figure 4, space autofocusing confocal laser image Raman optical spectrum detecting method, concrete method of testing includes following Step:
Laser instrument 610 in excitation beam system 600 produces exciting light, dissipates through minus lenses 620 and expands, then through the One condenser lenss 630 collimation becomes collimated light beam.
Collimated light beam reflects through dichroic optical system 900, and dissipate and look in the distance after focusing lens 110 of looking in the distance collecting lens 130 After focusing, it is radiated on sample 140, and Rayleigh scattering light is inspired on sample and is loaded with sample spectral characteristic Raman diffused light.
The Raman diffused light inspiring and Rayleigh scattering light collecting lens 130 of being looked in the distance collects recovering light path, through focusing of looking in the distance Compression light beam bore after mirror 110, after dichroic optical system 900 transmission, Raman diffused light and part Rayleigh scattering light first Beam splitting system 150 light splitting.
Part Rayleigh scattering light is reflected into the second beam splitting system 160, and wherein partly light is transmitted entrance confocal detection System 170, assembles through the 4th condenser lenss 200, through the second pin hole 190 transmission, forms Intensity response letter on the first detector 180 Number;Another part light is reflected into image sensing 310, and imaged lens 330 are imaged on imageing sensor 320.
Picture signal on the Intensity response signal of the first detector 180 and imageing sensor 320 is simultaneously sent to data Processing module 340, is sent to computer control system 350 after being then processed, after computer control system 350 is processed, formed Control signal simultaneously sends data processing module 340 to, and data processing module 340 produces focusing control signal and controls focusing of looking in the distance Mechanism 120 is focused, and the signal of the first detector 180 also can follow the tracks of change simultaneously, forms new control circulation, this process Continue, until peak response in the first detector 180, focus adjusting mechanism 120 completes the focusing of exciting light, now Raman dissipates Penetrate light transmission entrance spectrum investigating system 220 and carry out spectrographic detection, image sensing 310 completes sample image information gathering.
Utilization space autofocusing confocal laser image Raman spectral detection device, makes focusing by confocal detection response curve Mechanism 120 completes the focusing of exciting light, and now Raman diffused light is transmitted into spectrum investigating system 220 and carries out spectrographic detection, draws Graceful scattered light is assembled entrance the first spectrogrph 250 by the 5th condenser lenss 240, and Raman diffused light is through entrance slit 260, plane reflection Mirror 270 and the first concave reflection condenser lenss 280 reflection after reach spectrum grating 300, light beam after spectrum grating 300 diffraction, Second detector 230 is focused on by the second concave reflection condenser lenss 290 reflection.Due to grating diffration effect, in Raman spectrum The light of different wave length is separated from each other, from spectrogrph go out shoot out be sample Raman spectrum.
The first detector in measurement process, when spacescan is carried out to sample 140, in confocal detection system 170 180, the intensity response recording reaction sample 140 distance change is that (α, β, l), by gained intensity response I, (α, β l) pass I Deliver to data processing module 340 to be processed.
What in Raman spectroscopic detection system 220, the second detector 230 detected is loaded with drawing of sample 140 spectral information Graceful scattered light spectral signal is I (λ), and wherein λ is wavelength.
Image sensing acquisition sample space image information P (α, β, l).
By I (λ), (α, β, l) are sent to computer control system 350 and carry out data processing I, thus obtain comprising detected sample Product 140 positional information I (α, β, l) three-dimensional measurement information I (α, β, l, λ) with spectral information I (λ).
To sample 140 along α, to scanning, along l to scanning, repeat the above steps record right focus adjusting mechanism 120 of looking in the distance β Answer near objective focus positions one group of n comprise positional information [α, β, l] and I (λ) sequence measuring information [I (λ), α, β, l].
Using the corresponding positional information [α, β, l] of distinguishable region δ n, find out the spectral information In (λ) in corresponding δ n region Value, further according to the relation with space coordinatess [α, β, l], the information of reconstruct reflection measured object microcell δ n three dimensional structure and spectral characteristic In (α n, β n, ln, λ n), that is, achieve spectrographic detection and the three-dimensional geometry position sensing of microcell δ min.
The three dimension scale of corresponding minimum distinguishable region δ min and spectral characteristic are determined by following formula:
Achieve space autofocusing confocal laser image Raman spectrographic detection.
Microcell collection of illustrative plates is imaged
Iσmin(α, β, l)=In(α, β, l) 3D shape imaging
Iσmin(α, β, l)=In(λ) spectral measurement
Figure 4, it is seen that by the maximum point of confocal detection system 170 response curve 210, can accurately capture sharp The focal position of luminous speckle, from measurement sequence data, extracts the excitation spectrum of corresponding focus positions O, that is, achieves sample Spectrographic detection and three-dimensional geometry position sensing and image information.
As shown in figure 4, space autofocusing confocal laser image Raman spectral detection device is included positioned at dichroic light splitting system Unite the laser beam system 600 of 900 reflection directions;It is sequentially placed along light path positioned at dichroic optical system 900 transmission direction Look in the distance focusing lens 110, look in the distance collecting lens 130 and sample 140;Positioned at dichroic optical system 900 transmission direction first Beam splitting system 150;Raman spectroscopic detection system 220 positioned at the first beam splitting system 150 transmission direction;Positioned at the first beam splitting system Second beam splitting system 160 of 150 reflection directions;Confocal detection system 170 positioned at the second beam splitting system 160 transmission direction;Position In the second beam splitting system 160 reflection direction image sensing 310 and with confocal detection system 170, image sensing 310th, the data processing module 340 of the connection of Raman spectroscopic detection system 220 and focus adjusting mechanism 120 of looking in the distance and with data processing mould The computer control system 350 that block 340 is connected by serial ports.
Wherein, excitation beam system 600 is used for producing excitation beam, including the laser instrument 610 being sequentially placed along light path, bears Lens 620 and the first condenser lenss 630.
The 5th condenser lenss 240 that Raman spectroscopic detection system 220 includes being sequentially placed along light path, it is located at the 5th condenser lenss First spectrogrph 250 of 240 focal positions and the second detector 230 after the first spectrogrph 250.
Wherein, the first spectrogrph 250 includes entrance slit 260, the plane mirror 270, first being sequentially placed along light path Concave reflection condenser lenss 280 and spectrum grating 300.
Image sensing 310 includes imageing sensor 320, imaging len 330 and the second concave reflection condenser lenss 290; Confocal detection system 170 includes the 4th condenser lenss 200, is located at the second pin hole 190 and first of the 4th condenser lenss 200 focal position Detector 180.
Data processing module 340 and computer control system 350, the data collecting for fusion treatment simultaneously produces control Signal, and obtain image information.
Embodiment 2
Fig. 5 is space of the present invention autofocusing confocal laser image Raman optical spectrum detecting method and device embodiment 2 schematic diagram. The difference that the present embodiment is compared to embodiment 1 is:As shown in figure 5, excitation beam system 600 is placed in dichroic light splitting The transmission direction of system 900, focusing system 100 of looking in the distance is placed in the transmission direction of dichroic optical system 900, the first beam splitting system 150 reflection directions being placed at dichroic optical system 900.
Above in association with accompanying drawing, the specific embodiment of the present invention is described, but these explanations can not be understood to limit The scope of the present invention, protection scope of the present invention is limited by appended claims, any in the claims in the present invention base The change carrying out on plinth is all protection scope of the present invention.

Claims (9)

1. a kind of space autofocusing confocal laser image Raman spectral detection device includes excitation beam system, focusing system of looking in the distance System, dichroic optical system, the first beam splitting system, the second beam splitting system, Raman spectroscopic detection system, confocal detection system and figure As sensor-based system and data processing module;
Wherein, described excitation beam system and focusing system of looking in the distance are sequentially placed into the reflection side of dichroic optical system along light path To;
Described first beam splitting system is in the transmission direction of dichroic optical system;Raman spectroscopic detection system is located at the first light splitting The transmission direction of system;Second beam splitting system is located at the reflection direction of the first beam splitting system;Confocal detection system is located at second point The transmission direction of photosystem;Image sensing is located at the reflection direction of the second beam splitting system;
Described data processing module and Raman spectroscopic detection system, image sensing, confocal detection system and focusing system of looking in the distance System connects.
2. a kind of space according to claim 1 autofocusing confocal laser image Raman spectral detection device, its feature exists In described excitation beam system also includes light polarization modulator and iris filter.
3. a kind of space according to claim 2 autofocusing confocal laser image Raman spectral detection device, its feature exists In described iris filter is located between Polarization Controller and dichroic optical system or is located at dichroic optical system and prestige Between remote focusing system.
4. a kind of space according to claim 1 autofocusing confocal laser image Raman spectral detection device, its feature exists In, described excitation beam system is also sequentially placed into the transmission direction of dichroic optical system with focusing system of looking in the distance along light path, the One beam splitting system is placed in the reflection direction of dichroic optical system, and Raman spectroscopic detection system is located at the transmission of the first beam splitting system Direction, the second beam splitting system is located at the reflection direction that confocal detection system is located at the first beam splitting system, and confocal detection system is located at The transmission direction of the second beam splitting system, image sensing is located at the reflection direction of the second beam splitting system.
5. a kind of space according to claim 1 autofocusing confocal laser image Raman spectral detection device, its feature exists In described Raman spectroscopic detection system is common Raman spectroscopic detection system, including the 5th optically focused being sequentially placed along light path Mirror, the first spectrogrph positioned at the 5th condenser lenss focal position and the second detector after the first spectrogrph, for tested The detection of the surface spectrum of sample;Or confocal Raman spectra detection system, including the 7th condenser lenss being sequentially placed along light path, Positioned at the 3rd pin hole of the 7th condenser lenss focal position, the 8th condenser lenss after the 3rd pin hole, positioned at the 8th condenser lenss Jiao Second spectrogrph of point position and the 3rd detector after the second spectrogrph, for improving system signal noise ratio and spatial discrimination Power, completes the spectrographic detection to sample.
6. a kind of space according to claim 1 autofocusing confocal laser image Raman spectral detection device, its feature exists In described data processing module includes confocal data processing module, data fusion module data, control module and image module.
7. the space autofocusing confocal laser image Raman optical spectrum detecting method described in a kind of utilization claim 1, methods described Comprise the steps:
1) excitation beam system produces excitation beam, after dichroic optical system and focusing system of looking in the distance, is radiated at tested On sample, and inspire Rayleigh scattering light and the Raman diffused light being loaded with sample spectra characteristic;
2) Rayleigh scattering light in corresponding sample region and Raman diffused light again pass by focusing system of looking in the distance, focusing of being looked in the distance System is shaped to directional light, through dichroic optical system transmission simultaneously dichroic optical system to Rayleigh scattering light and Raman scattering Light carries out separating;
3) part Rayleigh scattering light, by dichroic optical system transmission, reflects and saturating through the second beam splitting system through the first beam splitting system Inject confocal detection system, using the first detector in confocal detection system, record the intensity of reflection sample position information Response I [α, β, l];
4) pass through focus adjusting mechanism, make the response of confocal detection system maximum, complete excitation beam and be autofocusing on sample, simultaneously Obtain the positional information [α, β, l] of sample;
5) complete after excitation beam focuses, image sensing obtain sample space image information P (α, β, l);
6) Raman diffused light, through dichroic optical system transmission, is transmitted into Raman spectroscopic detection system through the first beam splitting system, Record the Raman scattering signal I (λ) being loaded with sample characteristic using Raman spectroscopic detection system, carry out spectrum test;
7) I (λ) is sent to data processing module and carries out data processing, thus obtain comprising sample corresponding region position Spectral information I (λ) and object location information [α, β, l];
8) to sample along α, β scanning direction, focusing system of looking in the distance carries out focusing along l scanning direction, and repeat the above steps are surveyed One group of n that objective focus positions must be corresponded to comprise the sequence measuring information of positional information [α, β, l] and I (λ) [I (λ), α, β, L] and Pn (α, β, l);
9) utilize the corresponding positional information [α, β, l] of distinguishable region δ n, find out spectral information In (λ) value in corresponding δ n region, Further according to the relation with space coordinatess [α, β, l], information In of reconstruct reflection measured object microcell δ n three dimensional structure and spectral characteristic (α n, β n, ln, λ n);
10) three dimension scale of corresponding minimum distinguishable region δ min and spectral characteristic are determined by following formula:
8. a kind of space according to claim 7 autofocusing confocal laser image Raman optical spectrum detecting method, its feature exists In, the corresponding focusing system focal position of looking in the distance of confocal curves maximum value position, focused spot size is minimum herein, the region of detection Minimum, the out of focus region of the corresponding focusing system of looking in the distance of confocal curves other positions, the focal beam spot before Jiao or in defocused region Size increases with defocusing amount and increases.
9. a kind of space according to claim 7 autofocusing confocal laser image Raman optical spectrum detecting method, its feature exists In described excitation beam is polarized beam:Linear polarization, circular polarization or radial polarisation light or generated by pupil filtering technology Structure light beam.
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