CN104597034A - Raman spectra measuring device for multi-wavelength laser frequency shift excitation - Google Patents
Raman spectra measuring device for multi-wavelength laser frequency shift excitation Download PDFInfo
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Abstract
The invention relates to a Raman spectra measuring device for multi-wavelength laser frequency shift excitation. The Raman spectra measuring device comprises at least two semiconductor lasers used for generating lasers with fixed wavelengths, at least two laser control modules used for controlling parameters of the lasers, a fiber-optical switch used for switching light sources of the semiconductor lasers, a Raman probe used for carrying out focusing excitation on a sample to be detected and collecting Raman optical signals generated by the sample, a sample cell used for placing the sample to be detected, a grating splitting module used for dividing the Raman optical signals on space, a spectral data acquisition module used for carrying out photoelectric conversion on the Raman optical signals and an arithmetic processing module used for analyzing and extracting Raman spectra. The Raman spectra measuring device is capable of overcoming the disadvantages of the common tunable laser of instability, complex device or the like, effectively inhibiting fluorescence interference and realizing Raman spectra detection under strong fluorescence background.
Description
Technical field
The present invention relates to exact instrument and detection technique field.
Background technology
Raman spectroscopy has the outstanding advantages such as detection speed is fast, Non-Destructive Testing, is a kind of powerful tool carrying out molecular structure research, is widely used in the fields such as food security, biomedicine, public safety.But, often disturb with hyperfluorescenceZeng Yongminggaoyingguang in the Raman spectrum testing process of most of food, medicine, drugs.For hyperfluorescenceZeng Yongminggaoyingguang sample, fluorescence can flood Raman signal, and the existence of fluorescence has a strong impact on the identification of Raman spectrum characteristic peak, and therefore the Control of Fluorescence carried out in Raman spectrum testing process is necessary.In addition, the noise of the detector such as CCD itself and dark current also produce serious interference to the identification of some weak Raman peaks.
At present, free resolved Raman spectra method (the Matousek P of method of Fluorophotometry interference, Towrie M, StanleyA, et al.Efficient rejection of fluorescence from Raman spectra using picosecondKerr gating [J] .Applied Spectroscopy, 1999, 53 (12): 1485-1489), frequency domain filtering method (Mosier-Boss P A, Lieberman S H, Newbery R.Fluorescence rejection in Ramanspectroscopy by shifted-spectra, edge detection, and FFT filtering techniques [J] .Applied spectroscopy, 1995, 49 (5): 630-638), polynomial fitting method (Zhao J, Lui H, McLeanD I, et al.Automated autofluorescence background subtraction algorithm forbiomedical Raman spectroscopy [J] .Applied spectroscopy, 2007, 61 (11): 1225-1232), Wavelet Transform (Bertinetto C G, Vuorinen T.Automatic baseline recognitionfor the correction of large sets of spectra using continuous wavelet transformand iterative fitting [J] .Applied spectroscopy, 2014, 68 (2): 155-164) etc.The diversity of fluorescence background often causes the inefficacy of said method, and shift frequency excitation method (Shreve A P, Cherepy N J, MathiesR A.Effective rejection of fluorescence interference in Raman spectroscopy usinga shifted excitation difference technique [J] .Applied spectroscopy, 1992, 46 (4): 707-711) the general close excitation source of two wavelength that adopts detects sample, because the minor alteration of excitation wavelength is very little on fluorescence impact, wide band fluorescence background almost remains unchanged, and entirety occurs slight mobile by Raman signal.Therefore, as long as obtain Raman difference spectrum by subtracting each other after two original spectrum normalization, in Raman difference spectrum, fluorescence background is cancelled out each other, and only retains Raman signal, restrained effectively fluorescence interference.But Raman difference spectrum is not real Raman spectrum, need could restore the real Raman spectrum of sample by algorithm process.
Ensure that the stability of excitation source wavelength is the key factor of shift frequency excitation method.The excitation source of current shift frequency excitation method adopts tunable laser to obtain dual wavelength or multiwavelength laser usually.Tunable laser generally adopts Current Control Technology, temperature control technology or machine control techniques.The optical maser wavelength that the laser instrument of electric current or temperature-tunable produces is unstable, needs extra device to carry out lasting wavelength monitoring to ensure the stability of output wavelength.There are moveable parts in mechanically tunable laser device, is not suitable for being integrated in Portable Raman optical spectrum system.
Summary of the invention
The object of the present invention is to provide the raman spectroscopy measurement device that a kind of multiwavelength laser shift frequency excites, the interference of effective Fluorophotometry, the Raman spectrum realizing hyperfluorescenceZeng Yongminggaoyingguang background detects.
Technical scheme of the present invention is as follows:
The raman spectroscopy measurement device that multiwavelength laser shift frequency excites, is characterized in that: comprise the first laser instrument (1), second laser (2), the 3rd laser instrument (3), the first laser control module (4), second laser control module (5), the 3rd laser control module (6), fiber-optical switch (7), Raman probe (8), sample cell (10), grating beam splitting module (11), spectrum data gathering module (12), algorithm processing module (13);
Wherein the first laser instrument (1), second laser (2) and the 3rd laser instrument (3) comprise a laser emitting source, a thermistor temperature sensor, semiconductor refrigerator, a photodetector respectively;
The electric current that first laser instrument driver module (4) is connected for arranging the first laser instrument (1) with the laser emitting source (1-1) of the first laser instrument (1), the temperature that first laser instrument driver module (4) is connected for monitoring the first laser instrument (1) with the thermistor temperature sensor (1-2) of the first laser instrument (1), first laser instrument driver module (4) is connected with the semiconductor cooler (1-3) of the first laser instrument (1) for heating the first laser instrument (1) or freezing, the power that first laser instrument driver module (4) is connected for gathering the first laser instrument (1) with the photodetector (1-4) of the first laser instrument (1),
The electric current that second laser driver module (5) is connected for arranging second laser (2) with the laser emitting source (2-1) of second laser (2), the temperature that second laser driver module (5) is connected for monitoring second laser (2) with the thermistor temperature sensor (2-2) of second laser (2), second laser driver module (5) is connected with the semiconductor cooler (2-3) of second laser (2) for heating second laser (2) or freezing, the power that second laser driver module (5) is connected for gathering second laser (2) with the photodetector (2-5) of second laser (2).
The electric current that 3rd laser instrument driver module (6) is connected for arranging the 3rd laser instrument (3) with the laser emitting source (3-1) of the 3rd laser instrument (3), the temperature that 3rd laser instrument driver module (6) is connected for monitoring the 3rd laser instrument (3) with the thermistor temperature sensor (3-2) of the 3rd laser instrument (3), 3rd laser instrument driver module (6) is connected with the semiconductor cooler (3-3) of the 3rd laser instrument (3) for heating the 3rd laser instrument (3) or freeze, the power that 3rd laser instrument driver module (6) is connected for gathering the 3rd laser instrument (3) with the photodetector (3-5) of the 3rd laser instrument (3).
First laser instrument (1) and fiber-optical switch (7) pass through Fiber connection, second laser (2) and fiber-optical switch (7) pass through Fiber connection, laser instrument (3) and fiber-optical switch (7) pass through Fiber connection, fiber-optical switch (7) and Raman probe (8) pass through Fiber connection, testing sample (9) is positioned in sample cell (10), Raman probe (8) and grating beam splitting module (11) pass through Fiber connection, grating beam splitting module (11) is connected for data feedback and optimum configurations with spectrum data gathering module (12), spectrum data gathering module (12) is connected for data feedback with algorithm processing module (13).
In the preferred embodiment, the emission wavelength of the first laser instrument (1) is for being 784.5nm, and the emission wavelength of second laser (2) is 785.0nm, and the emission wavelength of the 3rd laser instrument (3) is 785.5nm.
In the preferred embodiment, described laser control module (4) comprises driving constant current source and temperature control unit, and the temperature stability of laser instrument is 0.1 DEG C, and laser wavelength fluctuated as 8pm in 24 hours.
In the preferred embodiment, the CCD that described spectrum data gathering module (12) is inner to grating beam splitting module (11) freezes, and preferably, cryogenic temperature reaches-15 DEG C.
The using method of the raman spectroscopy measurement device that aforesaid a kind of multiwavelength laser shift frequency excites, comprises the steps:
1) testing sample (9) is placed on sample cell (10) inner;
2) the first laser instrument (1) control module (4), second laser (2) control module (5), the 3rd laser instrument (3) control module (6) is started, wait for that laser control module reaches design temperature and power, ensure the wavelength stability of the first laser instrument (1), second laser (2), the 3rd laser instrument (3); The CCD starting spectrum data gathering module (12) inner to grating beam splitting module (11) freezes, and waits the temperature of setting to be achieved;
3) the first laser instrument (1), second laser (2), the 3rd laser instrument (3) is switched by fiber-optical switch (7), be transferred to Raman probe (8) respectively, Raman probe (8) transmission Laser Focusing is irradiated on testing sample (9);
4) Raman probe (8) collects the Raman diffused light filtering Rayleigh scattering light that sample produces, and is transmitted through the fiber to grating beam splitting module (11);
5) grating beam splitting module (11) makes Raman diffused light spatially separate by wavelength, and focus on the light-sensitive surface of detector C CD, obtain spectrum electric signal through opto-electronic conversion, and carry out data acquisition by spectrum data gathering module (12);
6) spectroscopic data is transferred to algorithm processing module (13) by spectrum data gathering module (12).
7) step 3 is repeated)-6), obtain the Raman spectrum of sample under different wave length laser.Last algorithm processing module (13) carries out the algorithm process of being correlated with, and obtains the Raman spectrum of sample.
In aforesaid using method, preferably, the emission wavelength of the first laser instrument (1) is for being 784.5nm, and the emission wavelength of second laser (2) is 785.0nm, and the emission wavelength of the 3rd laser instrument (3) is 785.5nm.
Advantage of the present invention is as follows:
(1) the present invention is directed to the needs of high fluorescence background raman spectroscopy measurement, multiwavelength laser shift frequency is adopted to excite the method for raman spectroscopy measurement, multiple fixed wave length semiconductor laser based on Volume Bragg grating technology is as LASER Light Source, thus realize the shift frequency of sample under multiwavelength laser and excite, carry out raman spectroscopy measurement and algorithm process, can the interference of effective Fluorophotometry.
(2) noise and the dark current that the present invention is directed to the detectors such as CCD itself also produce serious interference to the identification of some weak Raman peaks, adopt highly sensitive slim area array CCD as the detector of spectrum data gathering, and it is freezed, reduce dark current noise, improve the Raman spectrum acquisition performance of device.
Therefore, the present invention adopts based on the fixed wave length semiconductor laser of Volume Bragg grating technology as LASER Light Source, and it has the advantages such as Wavelength stabilized, line width.The temperature of laser instrument and power are controlled, ensures the stability of its output wavelength, compact conformation, efficient.
Accompanying drawing explanation
Fig. 1 is the raman spectroscopy measurement apparatus structure composition diagram that multiwavelength laser shift frequency of the present invention excites.
Fig. 2 is the raman spectroscopy measurement result that the multiwavelength laser shift frequency of the present invention to certain sample excites.
(a) three raman spectroscopy measurement figure of exciting of wavelength laser shift frequency;
B Raman spectrogram that () algorithm process is extracted
In figure
1-first laser instrument
The thermistor temperature sensor of laser emitting source 1-2 first laser instrument of 1-1 first laser instrument
The photodetector of semiconductor cooler 1-4 first laser instrument of 1-3 first laser instrument
2-second laser
The thermistor temperature sensor of the laser emitting source 2-2 second laser of 2-1 second laser
The photodetector of the semiconductor cooler 2-4 second laser of 2-3 second laser
3-the 3rd laser instrument
The thermistor temperature sensor of laser emitting source 3-2 the 3rd laser instrument of 3-1 the 3rd laser instrument
The photodetector of semiconductor cooler 3-4 the 3rd laser instrument of 3-3 the 3rd laser instrument
4-first laser control module 5-second laser control module 6-the 3rd laser control module
7-fiber-optical switch 8-Raman probe 9-testing sample 10-sample cell
11-grating beam splitting module 12-spectrum data gathering module 13-algorithm processing module
Embodiment
See Fig. 1, apparatus of the present invention comprise the first laser instrument 1, second laser 2, the 3rd laser instrument 3, first laser control module 4, second laser control module 5, the 3rd laser control module 6, fiber-optical switch 7, Raman probe 8, testing sample 9, sample cell 10, grating beam splitting module 11, spectrum data gathering module 12, algorithm processing module 13.Wherein the first laser instrument 1 comprises laser emitting source 1-1, thermistor temperature sensor 1-2, semiconductor cooler 1-3, photodetector 1-4.Second laser 2 comprises laser emitting source 2-1, thermistor temperature sensor 2-2, semiconductor cooler 2-3, photodetector 2-4.3rd laser instrument 3 comprises laser emitting source 3-1, thermistor temperature sensor 3-2, semiconductor cooler 3-3, photodetector 3-4.
The first described laser instrument driver module 4 is connected with the laser emitting source 1-1 of the first laser instrument 1, for arranging the electric current of the first laser instrument 1; First laser instrument driver module 4 is connected with the thermistor temperature sensor 1-2 of the first laser instrument 1, for monitoring the temperature of the first laser instrument 1; First laser instrument driver module 4 is connected with the semiconductor cooler 1-3 of the first laser instrument 1, for heating the first laser instrument 1 or freeze; First laser instrument driver module 4 is connected with the photodetector 1-4 of the first laser instrument 1, for gathering the power of the first laser instrument 1.
Described second laser driver module 5 is connected with the laser emitting source 2-1 of second laser 2, for arranging the electric current of second laser 2, second laser driver module 5 is connected with the thermistor temperature sensor 2-2 of second laser 2, for monitoring the temperature of second laser (2); Second laser driver module (5) is connected with the semiconductor cooler 2-3 of second laser (2), for heating second laser 2 or freezing; Second laser driver module 5 is connected with the photodetector 2-5 of second laser 2, for gathering the power of second laser 2.
The 3rd described laser instrument driver module 6 is connected with the laser emitting source 3-1 of the 3rd laser instrument 3, for arranging the electric current of the 3rd laser instrument 3; 3rd laser instrument driver module 6 is connected with the thermistor temperature sensor 3-2 of the 3rd laser instrument 3, for monitoring the temperature of the 3rd laser instrument 3; 3rd laser instrument driver module 6 is connected with the semiconductor cooler 3-3 of the 3rd laser instrument 3, for heating the 3rd laser instrument 3 or freeze; 3rd laser instrument driver module 6 is connected with the photodetector 3-5 of the 3rd laser instrument 3, for gathering the power of the 3rd laser instrument 3.
First laser instrument 1 passes through Fiber connection with fiber-optical switch 7, second laser 2 and fiber-optical switch 7 pass through Fiber connection, 3rd laser instrument 3 passes through Fiber connection with fiber-optical switch 7, fiber-optical switch 7 and Raman probe 8 pass through Fiber connection, testing sample 9 is positioned in sample cell 10, Raman probe 8 and grating beam splitting module 11 pass through Fiber connection, grating beam splitting module 11 is connected for data feedback and optimum configurations with spectrum data gathering module 12, and spectrum data gathering module 12 is connected for data feedback with algorithm processing module 13.
Wherein, grating beam splitting module (11) is inner adopts highly sensitive slim area array CCD.
Utilize multiwavelength laser shift frequency to excite testing sample, carry out raman spectroscopy measurement, comprise the steps:
1) testing sample 9 is placed on sample cell 10 li;
2) the first laser control module 4, second laser control module 5, the 3rd laser control module 6 is started, wait for that laser control module reaches design temperature and power, ensure the wavelength stability of the first laser instrument 1, second laser 2, the 3rd laser instrument 3, wherein, the wavelength of the first laser instrument 1 is 784.5nm, the wavelength of second laser 2 is 785.0nm, and the wavelength of the 3rd laser instrument 3 is 785.5nm; The CCD starting spectrum data gathering module 12 pairs of grating beam splitting modules 11 li freezes, and waits the temperature of setting to be achieved.
3) by fiber-optical switch 7 switched laser device 1, laser instrument 2, laser instrument 3, be transferred to Raman probe 8 respectively, Raman probe 8 transmits Laser Focusing and is irradiated on testing sample 9.
4) Raman probe 8 collects the Raman diffused light filtering Rayleigh scattering light that sample produces, and is transmitted through the fiber to grating beam splitting module 11.
5) grating beam splitting module 11 makes Raman diffused light spatially separate by wavelength, and focuses on the light-sensitive surface of detector C CD, obtains spectrum electric signal through opto-electronic conversion, and carries out data acquisition by spectrum data gathering module 12.
6) spectroscopic data is transferred to algorithm processing module 13 by spectrum data gathering module 12.
7) step 3 is repeated)-6), the Raman spectrum of sample under different wave length laser can be obtained.Last algorithm processing module 13 carries out the algorithm process of being correlated with, and obtains the Raman spectrum of sample;
Described laser control module 4 is made up of driving constant current source, temperature control unit, and the temperature stability realizing laser instrument is 0.1 DEG C, and laser wavelength fluctuated as 8pm in 24 hours.The CCD of described spectrum data gathering module 12 pairs of grating beam splitting modules 11 li carries out refrigeration and can reach-15 DEG C.
In addition, Fig. 2 is measurement under apparatus of the present invention recommended work mode i.e. three wavelength laser shift frequencies excite and algorithm process result, Fig. 2 (a) for device is to the raman spectroscopy measurement result of certain sample, the Raman spectrogram that Fig. 2 (b) extracts for algorithm process.
Claims (7)
1. the raman spectroscopy measurement device that excites of multiwavelength laser shift frequency, is characterized in that: comprise the first laser instrument (1), second laser (2), the 3rd laser instrument (3), the first laser control module (4), second laser control module (5), the 3rd laser control module (6), fiber-optical switch (7), Raman probe (8), sample cell (10), grating beam splitting module (11), spectrum data gathering module (12), algorithm processing module (13);
Wherein the first laser instrument (1), second laser (2) and the 3rd laser instrument (3) comprise a laser emitting source, a thermistor temperature sensor, semiconductor refrigerator, a photodetector respectively;
The electric current that first laser instrument driver module (4) is connected for arranging the first laser instrument (1) with the laser emitting source (1-1) of the first laser instrument (1), the temperature that first laser instrument driver module (4) is connected for monitoring the first laser instrument (1) with the thermistor temperature sensor (1-2) of the first laser instrument (1), first laser instrument driver module (4) is connected with the semiconductor cooler (1-3) of the first laser instrument (1) for heating the first laser instrument (1) or freezing, the power that first laser instrument driver module (4) is connected for gathering the first laser instrument (1) with the photodetector (1-4) of the first laser instrument (1),
The electric current that second laser driver module (5) is connected for arranging second laser (2) with the laser emitting source (2-1) of second laser (2), the temperature that second laser driver module (5) is connected for monitoring second laser (2) with the thermistor temperature sensor (2-2) of second laser (2), second laser driver module (5) is connected with the semiconductor cooler (2-3) of second laser (2) for heating second laser (2) or freezing, the power that second laser driver module (5) is connected for gathering second laser (2) with the photodetector (2-5) of second laser (2),
The electric current that 3rd laser instrument driver module (6) is connected for arranging the 3rd laser instrument (3) with the laser emitting source (3-1) of the 3rd laser instrument (3), the temperature that 3rd laser instrument driver module (6) is connected for monitoring the 3rd laser instrument (3) with the thermistor temperature sensor (3-2) of the 3rd laser instrument (3), 3rd laser instrument driver module (6) is connected with the semiconductor cooler (3-3) of the 3rd laser instrument (3) for heating the 3rd laser instrument (3) or freeze, the power that 3rd laser instrument driver module (6) is connected for gathering the 3rd laser instrument (3) with the photodetector (3-5) of the 3rd laser instrument (3),
First laser instrument (1) and fiber-optical switch (7) pass through Fiber connection, second laser (2) and fiber-optical switch (7) pass through Fiber connection, laser instrument (3) and fiber-optical switch (7) pass through Fiber connection, fiber-optical switch (7) and Raman probe (8) pass through Fiber connection, testing sample (9) is positioned in sample cell (10), Raman probe (8) and grating beam splitting module (11) pass through Fiber connection, grating beam splitting module (11) is connected for data feedback and optimum configurations with spectrum data gathering module (12), spectrum data gathering module (12) is connected for data feedback with algorithm processing module (13).
2. the raman spectroscopy measurement device that excites of a kind of multiwavelength laser shift frequency as claimed in claim 1, it is characterized in that: the emission wavelength of the first laser instrument (1) is for being 784.5nm, the emission wavelength of second laser (2) is 785.0nm, and the emission wavelength of the 3rd laser instrument (3) is 785.5nm.
3. the raman spectroscopy measurement device that excites of a kind of multiwavelength laser shift frequency as claimed in claim 1, it is characterized in that: described laser control module (4) comprises driving constant current source and temperature control unit, the temperature stability of laser instrument is 0.1 DEG C, and laser wavelength fluctuated as 8pm in 24 hours.
4. the raman spectroscopy measurement device that excites of a kind of multiwavelength laser shift frequency as claimed in claim 1, is characterized in that: the CCD that described spectrum data gathering module (12) is inner to grating beam splitting module (11) freezes.
5. the raman spectroscopy measurement device that excites of a kind of multiwavelength laser shift frequency as claimed in claim 4, is characterized in that: described cryogenic temperature reaches-15 DEG C.
6. the using method of raman spectroscopy measurement device that excites of a kind of multiwavelength laser shift frequency as claimed in claim 1, comprises the steps:
1) testing sample (9) is placed on sample cell (10) inner;
2) the first laser instrument (1) control module (4), second laser (2) control module (5), the 3rd laser instrument (3) control module (6) is started, wait for that laser control module reaches design temperature and power, ensure the wavelength stability of the first laser instrument (1), second laser (2), the 3rd laser instrument (3); The CCD starting spectrum data gathering module (12) inner to grating beam splitting module (11) freezes, and waits the temperature of setting to be achieved;
3) the first laser instrument (1), second laser (2), the 3rd laser instrument (3) is switched by fiber-optical switch (7), be transferred to Raman probe (8) respectively, Raman probe (8) transmission Laser Focusing is irradiated on testing sample (9);
4) Raman probe (8) collects the Raman diffused light filtering Rayleigh scattering light that sample produces, and is transmitted through the fiber to grating beam splitting module (11);
5) grating beam splitting module (11) makes Raman diffused light spatially separate by wavelength, and focus on the light-sensitive surface of detector C CD, obtain spectrum electric signal through opto-electronic conversion, and carry out data acquisition by spectrum data gathering module (12);
6) spectroscopic data is transferred to algorithm processing module (13) by spectrum data gathering module (12);
7) step 3 is repeated) to step 6), obtain the Raman spectrum of sample under different wave length laser.Last algorithm processing module (13) carries out the algorithm process of being correlated with, and obtains the Raman spectrum of sample.
7. the using method of raman spectroscopy measurement device that excites of a kind of multiwavelength laser shift frequency as claimed in claim 6, it is characterized in that: the emission wavelength of the first laser instrument (1) is for being 784.5nm, the emission wavelength of second laser (2) is 785.0nm, and the emission wavelength of the 3rd laser instrument (3) is 785.5nm.
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