CN103439254B - A kind of point pupil confocal laser Raman spectra test method and device - Google Patents
A kind of point pupil confocal laser Raman spectra test method and device Download PDFInfo
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Abstract
The invention belongs to microspectrum technical field of imaging, confocal microscopy is combined with Raman spectrum Detection Techniques, relate to a kind of point of pupil confocal laser Raman spectra test method and device.The Rayleigh scattering light abandoned in utilizing confocal Raman spectra to detect builds a point pupil confocal micro imaging system, realizes high-resolution imaging and the detection of sample three-dimensional geometry position; Utilize " extreme point " of point pupil confocal micro imaging system to control spectrum investigating system and accurately can catch the Raman spectral information that object lens focal spot excites, what then realize " collection of illustrative plates unification " divides pupil confocal Raman spectra high-space resolution imaging and detection.The high-space resolution that the present invention is microcell three-dimensional geometry position and spectrum detects and provides new technological approaches, the fields such as physics, chemistry, biomedicine, material science, environmental science, petrochemical complex, geology, medicine, food, criminal investigation and jewelry calibrating can be widely used in, nondestructive appreciation and degree of depth spectral analysis can be carried out to sample.
Description
Technical field
The invention belongs to microspectrum technical field of imaging, confocal microscopy is combined with Raman spectrum Detection Techniques, relate to high-space resolution spectrum test formation method and the device of one " collection of illustrative plates unification ", for carrying out high-resolution test etc. to the microscopic spectrum of sample.
Background technology
Confocal laser Raman spectroscopy causes molecule (or lattice) to produce vibration by incident laser and loses (or acquisition) portion of energy, scattered light frequency is changed, by analyzing scattered light, come the component of sensing molecule, structure and relative content etc., confocal laser Raman spectroscopy is also called as molecular probe technology.This technology had both inherited the high-resolution tomography feature of confocal microscopy, spectral analysis can be carried out again to sample, it not only can the raman spectral signal of different microcell in the same aspect of observing samples, the Raman signal of the every aspect that the degree of depth is different in observing samples can also be distinguished, tomoscan is carried out to sample, thus is issued to the effect of carrying out " optical section " in the situation of not lesioned sample.Confocal laser Raman spectrum measuring technology is as a kind of basic fundamental means of extremely important material structure measuring and analysis, be widely used in the fields such as physics, chemistry, biomedicine, material science, environmental science, petrochemical complex, geology, medicine, food, criminal investigation and jewelry calibrating, nondestructive appreciation and degree of depth spectral analysis can be carried out to sample, meanwhile, the photoluminescence research etc. of Sample Scan and low temperature analysis, material can also be carried out.
As shown in Figure 1, light-source system sends excitation beam through after polarization spectroscope, quarter-wave plate and focusing objective len to the principle of conventional confocal Raman spectrum tester, focuses on sample, inspires the Raman diffused light being loaded with sample spectra characteristic; Sample is moved by 3 D scanning system, the Raman diffused light of corresponding sample zones of different is made again to be polarized dichroic mirror by quarter-wave plate, the light that polarization splitting prism reflects is assembled by the first condenser, and utilization is positioned at the first pin hole Raman spectrum detection system below and records the raman scattering spectrum being loaded with sample spectral information.
Because the Raman diffused light inspired is very faint, existing confocal laser Raman spectrum detection instrument is in order to reduce the energy loss of Raman diffused light, larger pin hole is chosen in system, usually between φ 150 μm ~ φ 200 μm, because system utilizes photon excited to carry out focus location, pinhole size directly affects the halfwidth of confocal axial location curve, and pinhole size causes more greatly system Focus accuracy to reduce, and namely reduces spatial resolution.And system can only carry out spectrographic detection, and pattern is single, limits its application.
Meanwhile, the Raman diffused light of sample scattering only has 10 of the Reyleith scanttering light beam intensity of reflection
-3~ 10
-6doubly, and existing confocal Raman spectra detection instrument only make use of faint Raman diffused light and carries out spectrographic detection and abandon and be better than Raman diffused light 10
3~ 10
6rayleigh light beam doubly, utilizes Raman diffused light directly to carry out imaging, there is the deficiency that system sensitivity is lower.
In addition, in order to the metrical information obtaining accurately, enrich, both needed the single-point Raman spectrum of long period to detect, and needed again to carry out the detection of multiple spot Raman spectrum during Raman spectrum imaging, therefore Raman spectrum imaging needs the longer time.But, comparatively large by the impact of environment temperature, vibration, air shake etc. in the long-time imaging process of instrument, easily make instrument system produce drift, thus cause sample location to be detected out of focus; Follow the tracks of and aligning ability because existing confocal Raman spectra Detection Techniques do not possess real-time focal point, thus in whole imaging process, cannot ensure that it excites the position of hot spot to be in objective focus positions, the actual hot spot that excites is much larger than object lens focused light focal spot, its result constrains the microminiaturization of detectable area, limits the microscopic spectrum detectivity of confocal Raman spectrometer device.
Summary of the invention
The object of the invention is the deficiency for overcoming prior art, point pupil confocal laser Raman spectra test method and the device that provide a kind of confocal microscopy and Raman spectrum to detect to combine.The Rayleigh scattering light abandoned in utilizing confocal Raman spectra to detect builds a point pupil confocal micro imaging system, realizes high-resolution imaging and the detection of sample three-dimensional geometry position; Utilize " extreme point " of point pupil confocal micro imaging system to control spectrum investigating system and accurately can catch the Raman spectral information that object lens focal spot excites, what then realize " collection of illustrative plates unification " divides pupil confocal Raman spectra high-space resolution imaging and detection.
The object of the invention is to be achieved through the following technical solutions.
A kind of point pupil confocal laser Raman spectra test method, comprises the following steps:
The pupil plane measuring object lens is placed illumination iris and collects pupil.Light-source system sends the excitation beam that can inspire Raman spectrum.After excitation beam transmission illumination pupil, measurement object lens, focus on sample, inspire the Raman diffused light being loaded with sample spectral characteristic, reflect Reyleith scanttering light simultaneously.Raman diffused light and Reyleith scanttering light are through measuring object lens and arriving dichroic optical system after collecting pupil, and dichroic optical system carries out harmless separation to Raman diffused light and Reyleith scanttering light.
Reyleith scanttering light through dichroic optical system reflection enters a point pupil confocal laser detection system.Divide pupil confocal laser detection system to utilize detector lateral excursion that the resonse characteristic of point pupil confocal microscope system can be made to produce the characteristic of phase shift, realize the detection to sample microcell geometric position.Detailed process is: process the detection hot spot that point pupil confocal laser detection system receiving Reyleith scanttering light obtains, obtain search coverage, record point pupil confocal laser response I (u) of the concavo-convex change of reflection sample, the three dimension scale tomography detection of high-space resolution can be carried out, wherein, u is axial normalization optical coordinate.
Meanwhile, Raman diffused light through dichroic optical system transmission enters Raman spectrum detection system, utilize " extreme point " of point pupil confocal laser response curve and measurement objective focus positions accurately this characteristic corresponding, by " extreme point " of point pupil confocal laser response curve accurately corresponding focus measuring object lens (5), accurately catching by finding " extreme point " spectral information exciting hot spot focal position, realizing the spectrographic detection of high-space resolution.Can carry out in real time sample in measuring process, accurately follow the tracks of focus, the attitude of 3 D scanning system is controlled to ensure that sample is in focal position all the time in whole measuring process by computer processing system, the error produced because the factor such as environment temperature and vibration causes sample out of focus can be suppressed, improve measuring accuracy.
When the detection hot spot only obtained point pupil confocal laser detection system receiving Reyleith scanttering light processes, the three dimension scale tomography detection of high spatial resolution can be carried out;
When the spectral signal only docking the Raman spectrum detection system acquisition of receiving Raman diffused light processes, the spectrographic detection of high-space resolution can be carried out;
When the spectral signal obtained point pupil confocal laser detection system receiving Reyleith scanttering light the detection hot spot obtained and the Raman spectrum detection system receiving Raman diffused light processes simultaneously, the microcell collection of illustrative plates tomography of high-space resolution can be carried out, realize point pupil confocal laser Raman spectrum high-space resolution imaging and detection to sample " collection of illustrative plates unification ".
Especially, in the methods of the invention, described illumination iris and collection pupil can be circle, D shape or other shapes.
In the methods of the invention, excitation beam can be the light beam such as linearly polarized light beam, circularly polarized light beam; It can also be the structure light beam generated by pupil filtering technology.Light beam and pupil filtering combine with technique can compress measurement focused spot size, improve the transverse resolution of point pupil confocal laser detection system.
The inventive method can detect scattering spectrums such as comprising fluorescence, Brillouin scattering, Compton scattering light.
In the methods of the invention, the coupling of the measurement object lens to different N A value can be realized by means of only computer system software process, and debug without the need to again carrying out any hardware to system.
The invention provides a kind of point of pupil confocal laser Raman spectroscopy test device, comprise the light-source system of generation excitation beam, measurement object lens, illumination iris, collection pupil, 3 D scanning system, displacement transducer, dichroic optical system, Raman spectrum detection system, point pupil confocal laser measuring system, data processing unit and computer processing system.
Wherein, illumination iris and collection pupil are placed on the pupil plane of measurement object lens.Sample is fixed on the objective table of three-dimensional scanner.Illumination iris and measurement object lens are placed on the beam exit direction of light-source system successively, illumination iris is coaxial with excitation beam, after dichroic optical system is placed on and collects pupil, Raman spectrum detection system is placed in the transmission direction of dichroic optical system, divide pupil confocal laser detection system on the reflection direction of dichroic light-dividing device, the data that data processing unit is used for Raman spectrum detection system, point pupil confocal laser detection system and displacement transducer collect carry out fusion treatment.Sample is fixed on the objective table of 3 D scanning system.Computer processing system is used for coordinating 3 D scanning system, Raman spectrum detection system, point pupil confocal laser detection system and data processing unit and controlling.
In apparatus of the present invention, data processing unit comprises division focal spot detection module, maximum value computing module, for the treatment of the detection hot spot that image capturing system obtains, obtains a point pupil confocal laser response curve, thus obtains the positional information of sample; Comprising data fusion module, for merging positional information I (u) and spectral information I (r), completing sample three-dimensionalreconstruction and Spectrum Data Fusion I (x, y, z, r).
In apparatus of the present invention, Raman spectrum detection system can be normal Raman spectroscopy detection system, comprise the first condenser placed successively along light path, the spectral detector being positioned at the first condenser focal position, and the detector after being positioned at spectral detector, for the top layer Raman spectrum detection of sample.It can also be confocal Raman spectra detection system, comprise the first condenser placed successively along light path, the first pin hole being positioned at the first condenser focal position, be positioned at the spectral detector after the first pin hole, and the detector after being positioned at spectral detector, can parasitic light outside effectively filtering focus, improve the signal to noise ratio (S/N ratio) of Raman spectrum detection.
In apparatus of the present invention, by increasing beam modulation system between light-source system and illumination iris, comprise the 4th condenser that the 3rd condenser placed successively along light path, the second pin hole being positioned at the 3rd condenser focus place and focus are positioned at the second pin hole place, excitation beam for sending light-source system is modulated, to obtain more high spatial resolution and better stimulation effect.
In apparatus of the present invention, by increasing image enhancement system, the Aili spot that enlarged image acquisition system detects, comprise place successively along light path second condenser lens, with the confocal image enhancement system of second condenser lens and be positioned at the image capturing system at image enhancement system focus place, to improve a point acquisition precision for pupil confocal laser measurement mechanism.
Beneficial effect
The inventive method contrast prior art has following innovative point:
1, the present invention will divide pupil confocal laser microtechnic and Raman spectrum Detection Techniques to organically combine, merge the high precision Objective rallying point position tracking ability of point pupil confocal laser microtechnic, confocal Raman spectra microscope can be made to detect the accurately corresponding minimum sample spectra characteristic exciting focal beam spot region, significantly improve the microscopical raman microspectroscopy spectroscopic probing capabilities of existing confocal Raman spectra, this is one of innovative point being different from existing Raman spectrum Detection Techniques;
2, utilize dichroic light-dividing device to carry out harmless separation to Reyleith scanttering light and Raman diffused light, Reyleith scanttering light enters a point pupil confocal laser detection system, and Raman diffused light enters Raman spectrum detection system, improves system Raman spectral detectivity.This is the innovative point two being different from existing Raman spectrum Detection Techniques;
3, owing to adopting the method for division focal spot to obtain signal, by changing the parameter of tiny area set on image detection system detection focal plane to mate the reflectivity of different samples, thus its application can be expanded; Can also can realize the coupling of the measurement object lens to different N A value by means of only computer system software process, and debug without the need to again carrying out any hardware to system, be conducive to the versatility realizing instrument.This is the innovative point three being different from existing Raman spectrum Detection Techniques;
4, the present invention just can realize point micro-detection of pupil confocal laser, the detection of confocal laser Raman spectrum, point pupil confocal laser Raman spectrum detection multi-mode switching and process by means of only software exchange process.This is the innovative point four being different from existing Raman spectrum Detection Techniques.
The inventive method has following features:
1, merge point pupil confocal laser Detection Techniques and Raman spectrum Detection Techniques, utilize the accurate location of point pupil confocal laser system focusing, carry out Focus tracking measurement, significantly improve the spatial resolution of Raman spectrum detection;
2, effectively taking into account of range ability and resolution characteristic can be realized, simultaneously by be arranged on focal spot get the parameter of tiny area, to mate the sample of different reflectivity, range of application is expanded;
3, system has point micro-detection of pupil confocal laser, the detection of confocal laser Raman spectrum and divides a pupil confocal laser Raman spectrum to detect three kinds of mode of operations, only needs the switching that just can be realized Three models by software operation;
4, the use of dichroic optical system strengthens the Raman spectrum received by Raman spectrum detection system, improve the signal to noise ratio (S/N ratio) of Raman spectrum detection, the microscopical microscopic spectrum detectivity of confocal Raman spectra is significantly improved, also can reduce the light intensity requirement of system to excitation source.
Accompanying drawing explanation
Fig. 1 is confocal Raman spectra formation method schematic diagram;
Fig. 2 is of the present invention point of pupil confocal laser Raman spectra test method schematic diagram;
Fig. 3 is that D shape divides pupil confocal laser Raman pupil to test schematic diagram;
Fig. 4 is of the present invention point of pupil confocal laser Raman spectroscopy test device schematic diagram;
Fig. 5 is point pupil confocal laser Raman spectroscopy test device schematic diagram with confocal spectroscopic detection system;
Fig. 6 is point pupil confocal laser Raman spectroscopy test device schematic diagram with beam modulation system;
Fig. 7 is point pupil confocal laser Raman spectroscopy test device schematic diagram with detection focal spot amplification system;
Fig. 8 is point pupil confocal laser Raman spectra test method and a device embodiment schematic diagram;
Fig. 9 is point pupil confocal laser response curve and the Raman spectrum response curve schematic diagram of point pupil confocal laser Raman spectra test method.
Wherein, 1-light-source system, 2-measures object lens, 3-illumination iris, 4-collects pupil, 5-sample, 6-dichroic optical system, 7-Raman spectrum detection system, 8-first condenser, 9-spectral detector, 10-detector, 11-divides pupil confocal laser detection system, 12-second condenser lens, 13-image capturing system, 14-detector region, 15-3 D scanning system, 16-displacement transducer, 17-data processing unit, 18-division focal spot detection module, 19-extreme value computing module, 20-data fusion module, 21-computer processing system, 22-first pin hole, 23-beam modulation system, 24-the 3rd condenser, 25-second pin hole, 26-the 4th condenser, 27-image enhancement system, 28-divides pupil confocal laser response curve, 29-Raman spectrum response curve, 30-polarization spectroscope, 31-quarter-wave plate, 32-focusing objective len.
Embodiment
Below in conjunction with drawings and Examples, the present invention is described in further detail.
A kind of point pupil confocal laser Raman spectra test method, comprises the following steps:
As shown in Figure 2, illumination iris 3 and collection pupil 4 are placed on the pupil plane of measurement object lens 2.Light-source system 1 sends excitation beam, after excitation beam transmission illumination pupil 3, measurement object lens 2, focuses on sample 5, inspires the Raman diffused light being loaded with sample 5 spectral characteristic, reflect Reyleith scanttering light simultaneously.Raman diffused light and Reyleith scanttering light are through measuring object lens 2 and arriving dichroic optical system 6 after collecting pupil 4.Dichroic optical system 6 pairs of Raman diffused lights and Reyleith scanttering light carry out harmless separation.The Reyleith scanttering light reflected through dichroic optical system 6 enters a point pupil confocal laser detection system 11, divide pupil confocal laser detection system 11 to utilize detector lateral excursion that the axial response family curve of point pupil confocal microscope system can be made to produce the characteristic of phase shift, realize the detection to sample 5 microcell geometric position.Raman diffused light through dichroic optical system 6 transmission enters Raman spectrum detection system 7, utilize " extreme point " of point pupil confocal laser response curve 28 and focal position accurately this characteristic corresponding, accurately catching by finding " extreme point " spectral information exciting hot spot focal position, realizing the Raman spectrum detection of high-space resolution.
A kind of point pupil confocal laser Raman spectroscopy test device, as shown in Figure 4, comprise light-source system 1, measurement object lens 2, illumination iris 3, collect pupil 4, dichroic optical system 6, Raman spectrum detection system 7, point pupil confocal laser detection system 11, displacement transducer 16, data processing unit 17,3 D scanning system 15 and computer processing system 21.
Wherein, illumination iris 3 and collection pupil 4 are placed on the pupil plane of measurement object lens 2.Illumination iris 3 and measurement object lens 2 are positioned in the excitation beam exit direction of light-source system 1 successively, and illumination iris 3 is coaxial with excitation beam; After dichroic optical system 6 is positioned at and collects pupil 4.
Raman spectrum detection system 7 is positioned in the transmission direction of dichroic optical system 6; Raman spectrum detection system 7 comprises the first condenser 8, spectral detector 9 and detector 10.Wherein, the test surface of spectral detector 9 is positioned at the focus place of the first condenser 8, after detector 10 is positioned at spectral detector 9.The spectral signal obtained is sent to data processing unit 17 by detector 10.
Pupil confocal laser detection system 11 is divided to be positioned at two on the reflection direction of beam splitting system 6; Divide pupil confocal laser detection system 11 to comprise second condenser lens 12 and image capturing system 13, wherein, the test surface of image capturing system 13 is positioned at the focus place of second condenser lens 12.
The information that data processing unit 17 obtains for the treatment of Raman spectrum detection system 7 and point pupil confocal laser detection system 11.Comprise division focal spot detection module 18, extreme value computing module 19, data fusion module 20.Three's incidence relation is: the Aili spot that image capturing system 13 collects by division focal spot detection module 18 carries out splitting and detecting, the signal obtained enters extreme value computing module 19 to carry out extreme value and asks for, and the sample measurement point positional information obtained enters data fusion module 20.Data fusion module 20 goes back the spectral signal that pick-up probe 10 obtains, and merges spectral information and positional information.
Computer processing system 21 is responsible for the running coordinating and control whole device.Computer processing system 21 is connected with displacement transducer 16,3 D scanning system 15, data fusion module 20; Image capturing system 13 is connected with division focal spot detection module 18, and data fusion module 21 is connected with detector 10;
Wherein, control 3 D scanning system 15 by computer processing system 21 and move sample 5, make zones of different Reyleith scanttering light and to should the Raman diffused light of region sample 5 by measuring object lens 2 and collecting pupil 4; Displacement transducer 16 for by the real time position feedback of sample 5 to computer processing system 21 for the motion of sample 5 provides foundation.
Especially, circular illumination pupil and collection pupil can be replaced with other shapes (as D shape, to form D shape and divide pupil confocal laser Raman spectra test method schematic diagram, as shown in Figure 3).
Especially, apparatus of the present invention can also comprise the first pin hole 22, form point pupil confocal laser Raman spectroscopy test device with confocal spectroscopic detection system, as shown in Figure 5.First pin hole is positioned at the focal position of the first condenser 8.
Especially, apparatus of the present invention can also increase beam modulation system 23 between light-source system 1 and illumination iris 3, as shown in Figure 6.The 4th condenser 26 that beam modulation system 23 comprises the 3rd condenser 24 placed successively along light path, the second pin hole 25 being positioned at the 3rd condenser 24 focus place, focus are positioned at the second pin hole 25 place, forms point pupil confocal laser Raman spectroscopy test device with beam modulation system.
Especially, apparatus of the present invention can also increase image enhancement system 27 in point pupil confocal laser detection system 11, for the Aili spot that enlarged image acquisition system 13 detects, as shown in Figure 7.Image enhancement system 27 and the confocal point of second condenser lens 12, and image capturing system 13 is positioned at image enhancement system 27 focus place, to improve a point acquisition precision for pupil confocal laser measurement mechanism.
Embodiment
In the present embodiment, dichroic optical system 6 is notch filter, and spectral detector 9 is Raman spectroscopy detector, and image capturing system 13 is CCD, and image enhancement system 27 is enlarging objective.
As shown in Figure 8, divide pupil confocal laser Raman spectrum detection process as follows:
First, the light-source system 1 be made up of laser instrument sends the exciting light that can inspire sample Raman spectrum, exciting light enters the second pin hole 25 after the 3rd condenser 24 is assembled becomes pointolite, after the 4th condenser 26 collimator and extender, forms parallel excitation beam.Excitation beam, after illumination iris 3, measurement object lens 2, focuses on sample 5, inspires the Raman diffused light being loaded with sample 5 spectral characteristic.
Then, control 3 D scanning system 15 by computer system 21 and move sample 5, make zones of different Reyleith scanttering light and to should the Raman diffused light of region sample 5 by measuring object lens 2 and collecting pupil 4, Rayleigh reflected light is separated with Raman diffused light by notch filter 6.
The Reyleith scanttering light reflected through notch filter 6 enters a point pupil confocal laser measuring system 11, enlarging objective 27 is entered after second condenser lens 12 is assembled, hot spot after being exaggerated is detected by CCD13, the hot spot that CCD13 detects enters division focal spot detection module 18, at detection focal spot center, tiny area search coverage 14 is set, the response recording this region is I (u), the response obtained is carried out extreme value by extreme value computing module 19 to be asked for, and obtains point " extreme point " for pupil confocal laser response curve 28; Divide " extreme point " of pupil confocal laser response curve 28 accurately corresponding with the focused spot of excitation beam, obtained the elevation information of sample surfaces by " extreme point " of response curve 28, the positional information that bound site displacement sensor 16 feeds back reconstructs the three-dimensional surface shape of sample 5.
Raman diffused light through notch filter 6 transmission enters Raman spectrum detection system 7, through the first condenser 8 and enter after being positioned in its focus the first pin hole 22 Raman spectroscopy detector 9 with and subsequent detector 10, record raman scattering spectrum response curve I (r) 29 being loaded with sample 5 spectral information, wherein the wavelength of r inspire to by sample 5 stimulated luminescence Raman diffused light.
Only process is carried out to point pupil confocal laser detection system 11 acquisition detection hot spot receiving Reyleith scanttering light and obtain search coverage 14 points of pupil confocal lasers response I (u), accurately catch by its " extreme point " focal position exciting hot spot, system can carry out the three dimension scale tomography of high-space resolution; When the spectral response I (r) 29 that the Raman spectrum detection system 7 that only Raman diffused light is received in docking obtains processes, system can carry out Raman spectrum detection; When spectral signal I (r) dividing pupil confocal laser response I (u) and the Raman spectrum detection system 7 of Raman diffused light to obtain simultaneously obtained the detector 11 receiving Reyleith scanttering light processes, system can carry out the microcell collection of illustrative plates tomography of high-space resolution, namely realizes " collection of illustrative plates unification " Effect on Detecting of the high-space resolution of sample 5 geometric position information and spectral information;
As shown in Figure 8, the light-source system 1 dividing pupil confocal laser Raman spectrum sniffer to comprise generation excitation beam, measurement object lens 2, illumination iris 3, collection pupil 4, sample 5, notch filter 6, Raman spectrum detection system 7, point pupil confocal laser measuring system 11,3 D scanning system 15, displacement transducer 16 and data processing unit 17, wherein, the pupil plane measuring object lens 2 be divided into illumination iris 3 and collect pupil 4, measurement object lens 2 and illumination iris 3 are placed on the beam exit direction of light-source system 1, illumination iris 3 is coaxial with excitation beam, after notch filter 6 is placed on and collects pupil 4, Raman spectrum detection system 7 is placed in the transmission direction of notch filter 6, pupil confocal laser measuring system 11 is divided to be placed on the reflection direction of notch filter 6, data processing unit 17 is for melting merging treatment Raman spectrum detection system 7, divide the data that pupil confocal laser detection system 11 and displacement transducer 16 collect, in Raman spectrum detection system 7, the focus place of the first condenser 8 places the first pin hole 22 pairs of parasitic lights and carries out filtering, to improve Raman spectrum detection efficiency, light-source system can by comprising the 3rd condenser 24, be positioned at the excitation beam that beam modulation system 23 pairs of light-source systems 1 that the 4th condenser 26 that second pin hole 25 at the 3rd condenser 24 focus place and focus be positioned at the second pin hole 25 place forms send and modulate, to improve the quality of excitation beam, the picture be arranged on image capturing system 11 second condenser lens 12 focal plane is amplified on the test surface of rear arrival CCD13 by Nonlinear magnify object lens 27, so that detector carries out division focal spot detection, improves detection accuracy.
Claims (7)
1. a point pupil confocal laser Raman spectra test method, is characterized in that, comprise the following steps:
The pupil plane measuring object lens (2) is placed illumination iris (3) and collects pupil (4); Light-source system (1) sends the excitation beam that can inspire Raman spectrum; Excitation beam successively transmission illumination pupil (3), measure object lens (2) after, focus on sample (5), inspire the Raman diffused light being loaded with sample spectral characteristic, reflect Reyleith scanttering light simultaneously; Raman diffused light and Reyleith scanttering light are successively through measuring object lens (2) and arriving dichroic optical system (6) after collecting pupil (4), and dichroic optical system (6) carries out harmless separation to Raman diffused light and Reyleith scanttering light;
The Reyleith scanttering light reflected through dichroic optical system (6) enters a point pupil confocal laser detection system (11); Divide pupil confocal laser detection system (11) to utilize detector lateral excursion that the axial response family curve of point pupil confocal microscope system can be made to produce the characteristic of phase shift, realize the detection to sample microcell geometric position; The detection hot spot that point pupil confocal laser detection system (11) receiving Reyleith scanttering light obtains is processed, obtain search coverage, record point pupil confocal laser response I (u) of the concavo-convex change of reflection sample, the three dimension scale tomography detection of high-space resolution can be carried out, wherein, u is axial normalization optical coordinate;
Meanwhile, Raman diffused light through dichroic optical system (6) transmission enters Raman spectrum detection system (7), utilize " extreme point " of point pupil confocal laser response curve and measurement object lens (2) focal position accurately this characteristic corresponding, by " extreme point " of point pupil confocal laser response curve accurately corresponding focus measuring object lens (5), accurately catching by finding " extreme point " spectral information exciting hot spot focal position, realizing the spectrographic detection of high-space resolution;
When the detection hot spot only obtained point pupil confocal laser detection system (11) receiving Reyleith scanttering light processes, the three dimension scale tomography detection of high-space resolution can be carried out;
When the spectral signal that the Raman spectrum detection system (7) of only docking receipts Raman diffused light obtains processes, the spectrographic detection of high-space resolution can be carried out;
When the spectral signal obtained point pupil confocal laser detection system (11) receiving Reyleith scanttering light the detection hot spot obtained and the Raman spectrum detection system (7) receiving Raman diffused light processes simultaneously, the microcell collection of illustrative plates tomography of high-space resolution can be carried out, realize point pupil confocal laser Raman spectrum high-space resolution imaging and detection to sample " collection of illustrative plates unification ".
2. according to claim 1 point of pupil confocal laser Raman spectra test method, is characterized in that: illumination iris (3) and collection pupil (4) are circular or D shape.
3. according to claim 1 point of pupil confocal laser Raman spectra test method, it is characterized in that: excitation beam is linearly polarized light beam or circularly polarized light beam, the combination of light beam and pupil filtering technology is compressed and is measured focused spot size, improves the transverse resolution of point pupil confocal laser detection system.
4. a point pupil confocal laser Raman spectroscopy test device, comprises light-source system (1), measurement object lens (2), dichroic optical system (6), Raman spectrum detection system (7), point pupil confocal laser detection system (11), displacement transducer (16), data processing unit (17), 3 D scanning system (15) and computer processing system (21); It is characterized in that: described proving installation also comprises illumination iris (3) and collects pupil (4), wherein, illumination iris (3) and collection pupil (4) are placed on the pupil plane of measurement object lens (2); Illumination iris (3) and measurement object lens (2) are positioned in the excitation beam exit direction of light-source system (1) successively, and illumination iris (3) is coaxial with excitation beam; After dichroic optical system (6) is positioned at and collects pupil (4); Sample (5) is fixed on 3 D scanning system (15) objective table;
Raman spectrum detection system (7) is positioned in the transmission direction of dichroic optical system (6); Raman spectrum detection system (7) comprises the first condenser (8), spectral detector (9) and detector (10); Wherein, the test surface of spectral detector (9) is positioned at the focus place of the first condenser (8), after detector (10) is positioned at spectral detector (9); The spectral signal obtained is sent to data processing unit (17) by detector (10);
Pupil confocal laser detection system (11) is divided to be positioned at two on the reflection direction of beam splitting system (6); Pupil confocal laser detection system (11) is divided to comprise second condenser lens (12) and image capturing system (13), wherein, the test surface of image capturing system (13) is positioned at the focus place of second condenser lens (12);
The information that data processing unit (17) obtains for the treatment of Raman spectrum detection system (7) and point pupil confocal laser detection system (11), data processing unit (17) comprises division focal spot detection module (18), extreme value computing module (19), data fusion module (20); Wherein, the Aili spot that image capturing system (13) collects carries out splitting and detecting by division focal spot detection module (18), the signal obtained enters extreme value computing module (19) to carry out extreme value and asks for, and the sample measurement point positional information obtained enters data fusion module (20); The spectral signal that data fusion module (20) pick-up probe (10) obtains, and merge spectral information and positional information;
Computer processing system (21) is responsible for the running coordinating and control whole proving installation; Computer processing system (21) is connected with displacement transducer (16), 3 D scanning system (15), data fusion module (20); Image capturing system (13) is connected with division focal spot detection module (18), and data fusion module (21) is connected with detector (10); Wherein, by computer processing system (21) control 3 D scanning system (15) mobile sample (5), make zones of different Reyleith scanttering light and to should region sample (5) Raman diffused light by measure object lens (2) and collection pupil (4); Displacement transducer (16) for by the real time position feedback of sample (5) to computer processing system (21).
5. a kind of point of pupil confocal laser Raman spectroscopy test device as claimed in claim 4, it is characterized in that, described proving installation also comprises first pin hole (22) of the focal position being positioned at the first condenser (8).
6. a kind of point of pupil confocal laser Raman spectroscopy test device as claimed in claim 4, is characterized in that, increase beam modulation system (23) between light-source system (1) and illumination iris (3); The 4th condenser (26) that beam modulation system (23) comprises the 3rd condenser (24) placed successively along light path, the second pin hole (25) being positioned at the 3rd condenser (24) focus place, focus are positioned at the second pin hole (25) place.
7. a kind of point of pupil confocal laser Raman spectroscopy test device as claimed in claim 4, it is characterized in that, image enhancement system (27) is increased, for the Aili spot that enlarged image acquisition system (13) detects in point pupil confocal laser detection system (11); Image enhancement system (27) and the confocal point of second condenser lens (12), and image capturing system (13) is positioned at image enhancement system (27) focus place.
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