CN104749135B - Mid-infrared light spectrometer - Google Patents
Mid-infrared light spectrometer Download PDFInfo
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- CN104749135B CN104749135B CN201510150145.4A CN201510150145A CN104749135B CN 104749135 B CN104749135 B CN 104749135B CN 201510150145 A CN201510150145 A CN 201510150145A CN 104749135 B CN104749135 B CN 104749135B
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Abstract
The invention discloses a kind of mid-infrared light spectrometer that nanoseconds resolution is realized using temperature-jump, the dynamic structure for measuring protein changes.The mid-infrared light spectrometer includes：Wideband is tunable CO laser, for sending middle infrared acquisition laser to testing protein solution；Nanosecond adjusts Q holmium lasers, for sending pumping laser to heat testing protein solution；Pump probe light path system, for detecting the intensity of the middle infrared acquisition laser after passing through testing protein solution；And data collecting system, the data of the intensity detected with collection from pump probe light path system.The transient state ir data and the dynamic information of protein dynamic structure change that the mid-infrared light spectrometer of the present invention can be changed with accurate measurement protein dynamic structure, the measurement of high-precision secondary protein structure dynamic change can be realized, ir data precision can reach 1 × 10‑4OD。
Description
Technical field
The present invention relates to Superfast time resolution middle infrared spectrum fields of measurement, more particularly, to a kind of mid-infrared light spectrometer.
Background technology
Protein science is 21 century human sciences science with fastest developing speed and most popular.Infrared spectrum technology is due to it
Include substantial amounts of structure of matter information, be widely used in protein science research.At present, using infrared spectrum skill
The structure change of art research protein mainly has two fields：The Stable structure change of protein and the dynamic structure of protein become
Change.The Stable structure change of infrared spectrum technology research protein depends on FTIS (FTIR).
It can provide the basic structure information of protein, but can not provide the dynamic structure information of protein.
In order to study the change of the dynamic structure of protein, people have developed reaction-stop method, and temperature-jump method and laser lure
The method for leading chemical reaction is used for the structure change for triggering protein.Reaction-stop method has the dead time of at least musec order,
It is extremely difficult to the time resolution of nanosecond order.And the method for laser induced chemical reaction is used for, it is necessary to be added in protein solution
The material of laser excitation, there is certain damage to protein.And laser pulse temperature-raising method is because of a coctoprotein solution and to egg
White matter is not almost damaged, and becomes the method for relatively having advantage of inducible protein change.Laser technology is limited to, it is current quick
Heating-up time differentiates infrared spectrum technology, more using tune Q Nd:The 1064nm laser of YAG laser output passes through high pressure hydrogen pond
Produce 1900nm laser method come produce heavy water heating clock.This method is due to needing by a Raman frequency shift
Non-linear process, cause caused by 1900nm laser, optical quality is poor, and spatial jitter is very big, system
Long-term working stability is also poor.
The content of the invention
The purpose of the present invention aims to provide a kind of middle infrared spectrum that temperature-jump can be utilized to realize nanoseconds resolution
Instrument, it is possible to achieve the measurement of high-precision secondary protein structure dynamic change.
In order to realize foregoing invention purpose, the invention provides a kind of mid-infrared light spectrometer, for measuring the dynamic of protein
State structure change, the mid-infrared light spectrometer include：Wideband is tunable CO laser, for being sent out to testing protein solution
Go out middle infrared acquisition laser；Nanosecond adjusts Q holmium lasers, for sending pumping laser to heat testing protein solution；Pumping is visited
Light path system is surveyed, for detecting the intensity of the middle infrared acquisition laser after passing through testing protein solution；And data acquisition system
System, for gathering the data of the intensity detected from pump probe light path system；The pumping that nanosecond adjusts Q holmium lasers to send swashs
The spot diameter that light is formed on sample cell is 2mm.
Further, pump probe light path system includes light path part, detector, amplifier and the filtering set gradually
Device.
Further, detector is liquid nitrogen refrigerating type high speed mercury-cadmium tellurid detector；Amplifier is DC-100MHz amplifiers.
Further, the output wavelength of the tunable CO laser of wideband is 5.2 μm~6.2 μm.
Further, the output pulse width for adjusting Q holmium lasers is 50ns, output wavelength 2080nm.
Further, the mid-infrared light spectrometer also includes being used for the sample cell for placing testing protein solution, sample cell tool
There is the second area of the first area for placing reference heavy water and the heavy aqueous solution for placing the solution containing testing protein.
Further, sample cell is rotary structure, and can be connect by its rotation first area and second area
The pumping for receiving the tunable CO laser of the wideband middle infrared acquisition laser sent and nanosecond adjusting Q holmium lasers to send swashs
Light.
Wideband is tunable, and middle infrared acquisition laser that CO laser sends forms the spot diameter on sample cell
For 0.4mm.
Further, data collecting system includes 8 high-speed collection cards, and minimum is distinguished as 0.36mV.
Further, data collecting system is arranged to the down-sampled type collection data with nanosecond to second time frame.
Beneficial effects of the present invention：Mid-infrared light spectrometer is differentiated using the nanosecond pulse heating-up time provided by the invention, can
Believed with the dynamics of the transient state ir data of accurate measurement protein dynamic structure change and the change of protein dynamic structure
Breath, its ir data precision can reach 1 × 10-4OD.Wherein, using high stability low temperature flow formula wideband tunable one
Carbon oxide laser device is as detection optical detection Secondary structure information, it is possible to achieve wide mid-infrared light spectrometry, it is complete
The absorption of all standing secondary protein structure acid amides I '.Q holmium lasers are adjusted using nanosecond short pulse, pumping laser can be used as straight
The heating that nanoseconds are carried out to the heavy aqueous solution containing protein is connect, there is the longitudinal direction heated to heavy aqueous solution uniformly, laterally
The advantages that uniformly and prolonged stabilization heats.Pump probe process to sample is realized using pump probe light path system,
It is stable with easy to operate and system, effectively reduce the advantages such as veiling glare.Realize that nanoseconds arrive using data collecting system
The high-speed data acquisition of time second.
According to the accompanying drawings will be brighter to the detailed description of the specific embodiment of the invention, those skilled in the art
Above-mentioned and other purposes, the advantages and features of the present invention.
Brief description of the drawings
Some specific embodiments of the present invention are described in detail by way of example, and not by way of limitation with reference to the accompanying drawings hereinafter.
Identical reference denotes same or similar part or part in accompanying drawing.It should be appreciated by those skilled in the art that these
What accompanying drawing was not necessarily drawn to scale.In accompanying drawing：
Fig. 1 is the structural frames of the nanosecond pulse heating-up time resolution mid-infrared light spectrometer employed in the embodiment of the present invention
Figure；
Fig. 2 is the tunable carbon monoxide of high stability low temperature flow formula wideband employed in invention embodiment
The structural representation of laser；
Fig. 3 is the structural representation that the nanosecond short pulse of exemplary embodiments of the present invention adjusts the optics cavity of Q holmium lasers；
Fig. 4 is the long-time result schematic diagram of the data acquisition by the way of down-sampled in the embodiment of the present invention；
Heavy aqueous solution is in 1645cm when Fig. 5 is moment heating in the embodiment of the present invention-1The absorbance dynamic changing curve at place
Schematic diagram；
Fig. 6 is that heavy aqueous solution and heavy aqueous solution containing Cyt C protein exist when moment heats up in the embodiment of the present invention
1645cm-1The absorbance dynamic changing curve figure at place；
Fig. 7 is heavy aqueous solution in the embodiment of the present invention in 1645cm-1The dynamic accuracy schematic diagram of the absorbance at place；
Cyt C protein is in 1645cm when Fig. 8 is moment heating in the embodiment of the present invention-1The absorbance dynamic change at place
Schematic diagram；And
Fig. 9 is the transient state spectral schematic of the cromoci after 2 microseconds of moment heating in the embodiment of the present invention.
Embodiment
As shown in figure 1, the mid-infrared light spectrometer provided by the invention that nanoseconds resolution can be realized using temperature-jump
Mainly include the tunable CO laser 10 of wideband, nanosecond short pulse adjusts Q holmium lasers 20, pump probe light path system and
Data collecting system 40.The transient state infrared light that the mid-infrared light spectrometer of the present invention can be changed with accurate measurement protein dynamic structure
Modal data and the dynamic information of protein dynamic structure change, its ir data precision can reach 1 × 10-4OD。
Each part of centering infrared spectrometer describes in detail separately below：
High stability low temperature flow formula wideband is tunable CO laser 10 is used for red in being sent to testing protein
Outer exploring laser light, to detect the secondary structure information of testing protein, it is possible to achieve wide mid-infrared light spectrometry, be completely covered
The absorption of secondary protein structure.Specifically, the tunable CO laser 10 of the wideband is solid using containing for liquid nitrogen cooling
Surely the mixed gas of the helium, nitrogen, carbon monoxide and the oxygen that match is as laser medium, the proportioning of the gas wavelength needed for
The difference of scope is adjusted.As shown in Fig. 2 during work, the gas mixed via electronic flowmeter enters quartzy tube passage.
One end that quartzy tube passage connects quartzy helix tube is gas input, and the other end is the outlet side for being connected with vavuum pump.By electricity
Sub- valve control and vavuum pump (not shown) realize the steady flow of gas, and air pressure is about 26.5torr during work.Work gas
Body is cooled down by liquid nitrogen.
In terms of protein structure change is detected, it is used directly for detecting the infrared laser of Protein secondary structure, mainly
Have lead salt laser and carbon monoxide (CO) laser and it is most recently newly go out QCL.Relative to lead salt laser and
For QCL, carbon monoxide (CO) laser 10 exports with wide spectrum, can be completely covered the two of protein
The advantages of infrared indication of the band of level structure acid amides I '.
Fig. 2 is the basic structure of the tunable CO laser of high stability low temperature flow formula wideband.The wideband is adjustable
Humorous CO laser 10 is using energisation mode of the bidirectional high-pressure electric pump as laser.The concrete structure of high pressure electric pump
For：Laser cavity both ends are anodes, and two anodes share middle negative electrode.Voltage between anode and cathode is about 7KV.The one of laser cavity
The gold-plated concave mirror for r=5m is held, one end is Electronic control rotating shutter.Grating is combined perpendicular to the gold-plated of grating surface placement
Level crossing and corresponding light path design can be realized in grating rotation process, export the wavelength change of laser but export laser
Direction does not change.R=5m concave surface gold mirror end mirror, for being totally reflected to infrared light.The concave surface gold mirror is mounted in one
Can be with the mirror holder of electronic fine setting, in order to be finely adjusted to the optical cavity of laser.
The laser dexterously employs outgoing mirror of the grating as laser.1 grade of light of grating returns to laser cavity, and 0
Level light output, delivery efficiency is about 10%.In order that the direction of output light does not change, some are optically carried out and have set
Meter：Grating and a plate plane gold mirror are disposed vertically, and the two is fixed on same turntable.Grating and plane gold mirror are simultaneously
Rotated.The deviation in the direction of the light caused by the rotation of grating is returned by the plane gold mirror compensation rotated therewith, from
And ensure that the exit direction of light will not change.In one embodiment of the invention, use can carry out the beautiful jade of temperature control
Steel pipe come ensure laser chamber length stabilization.The air distribution system of laser, grating control and high voltage control are all by labview programs
Control electronics element is realized.
Using the tunable CO laser 10 of high stability low temperature flow formula wideband of the present invention, there is following property
Energy：1) in 1580cm-1~1980cm-1In the range of output energy be more than 10mw；2) wavelength interval is about 4cm-1；3) short time
Stability is one thousandth, illustrates that the laser output of tunable CO laser 10 is more stable；4) prolonged shake
＜ 20%, illustrate the steady in a long-term of its working air current and voltage.
Nanosecond, short pulse was when adjusting the Q holmium lasers 20 can be used as pump light to heavy aqueous solution progress nanosecond containing protein
Between heating, have the advantages that to the vertical and horizontal of heavy aqueous solution heating uniformly and prolonged stable heating, Ke Yizhi
Connect and temperature-jump is carried out to heavy water.Fig. 3 adjusts the structure chart of the optics cavity of Q Ho lasers 20 for nanosecond short pulse.Laser cavity is using flat
Flat cavity configuration, one end are total reflective mirror 21, and one end is the output coupling mirror 22 that delivery efficiency is 20%.Nanosecond short pulse adjust Q holmiums swash
Light device is used doped with 1.5%Cr, 5.8%Tm, the Cr of 0.35%Ho elements, Tm, Ho:YAG crystal dodges as laser crystal 23
Light lamp 24 is used as pumping source.Electro-optic Q-switched crystal is：La3Ga5SiO14(LGS) crystal 25, Q mode of operations are adjusted using pressure type is moved back.Rise
Inclined device is a pair of aluminum oxide (Al that Brewster's angle is placed2O3) crystal 26, modulate Depolarization with the quartz wave-plate 27 of λ/4.
The power supply (nothing is shown in the two figure) that flash generator and adjusting Q crystal use derives from business procurement.Laser crystal
Water cooling realizes that water cooling unit water cooling temperature is 9 during work by the high-power water-cooling machine (not shown) that temperature-controlled precision is 0.1 degree
℃.Being primarily due to nanosecond short pulse using water-cooling pattern adjusts the gain of laser crystal 23 of Q Ho lasers relatively low, it is necessary to larger
Pumping could realize that laser exports, temperature is higher when laser crystal 23 works, and easily forms heat aggregation, it is necessary to accurate control
Its temperature ensures the stabilization of laser work.In one embodiment of the invention, nanosecond short pulse adjusts Q Ho lasers 20
Service behaviour is as follows：Working frequency:3Hz, output wavelength 2080nm, output pulse width：50ns, export energy：20mj, energy stabilization
Property:About 1%.
In the exemplary embodiments of the present invention, in addition to for placing the sample cell 50 of testing protein.The sample
Pond can use the structure of the double samples in a pond.That is sample cell can have two regions, for placing reference heavy water
First area 51 and the second area 52 for placing the heavy aqueous solution containing testing protein.Sample cell is preferably rotary knot
Structure, so the tunable oxygen of wideband can be received by the rotation first area 51 of sample cell and second area 52
Change the middle infrared acquisition laser that carbon laser 10 is sent and the pumping laser for adjusting Q holmium lasers 20 to send nanosecond.Sample cell 50
Structure is：A polytetrafluoro thin slice with two circular holes, the thickness of polytetrafluoro thin slice are placed among the plain film of two blocks of calcirm-fluoride
For 50 microns, so as to realize that the thickness of sample is 50 microns.
In order to reach higher moment increasing extent of temperature, the laser that nanosecond adjusts Q Ho lasers 20 to export is carried out by lens
Focus on, it is 2mm to control spot size of the nanosecond short pulse Q-switch laser at sample.Meanwhile in order to allow exploring laser light to be completely in
In heated sample scope, and the metastable region of temperature is in, detection light is focused with short lens, spot diameter
Control is in 0.2mm.In a preferred embodiment of the invention, the pumping laser for adjusting Q holmium lasers 20 to send is formed in sample
Spot diameter on pond is 2mm, and wideband is tunable, and middle infrared acquisition laser that CO laser 10 sends is formed in sample
Spot diameter on pond 50 is 0.4mm.
Pump probe light path system is used for the intensity for detecting the middle infrared acquisition laser after passing through testing protein solution.Adopt
The pump probe process to protein example to be measured is realized with pump probe light path system, wherein, the pump probe light path system
Employed in all speculums be Jin Jing 32, all lens are calcium fluoride lens 33, using wiregrating 31 be used for adjust
Up to the detection light energy at sample cell 50.
In a preferred embodiment of the invention, pump probe light path system includes light path part, the spy set gradually
Survey device 34, amplifier 35 and wave filter 36.Wherein, detector 34 swashs to detect the middle infrared acquisition after passing through testing protein
The intensity of light.The preferred liquid nitrogen refrigerating type high speed mercury-cadmium tellurid detector of detector.Amplifier 35 is to centering infrared acquisition laser
Intensity is amplified.Amplifier is preferably DC-100MHz amplifiers.Wave filter 36 is to the middle infrared acquisition laser after amplification
Strength signal filtered, the electromagnetic interference in filtering appts and space, with the electrical signal of frequency needed for obtaining.
Using data collecting system 40 nanoseconds can be realized to the high-speed data acquisition of time second.In order to realize nanosecond
To the data acquisition of second-time, in a preferred embodiment of the invention, data collecting system 40 employs exponential drop
Sampling techniques, so both can guarantee that the high time resolution precision of short-time dynamics, and and can ensures prolonged dynamics data
Collection.Down-sampled data acquisition long-time result is as shown in Figure 4.Figure 4, it is seen that acquisition system is within 1 millisecond
Acquisition precision is higher, can obtain detailed dynamic variation information early stage, while realizes and adopted up to 10 milliseconds of data again
Collect scope, prolonged dynamic-change information can be obtained roughly, while and can ensures the smoothness of data acquisition.
Specifically, the mode of operation of pump probe light path system and data collecting system 40 is as follows：Liquid nitrogen refrigerating type is at a high speed
The amplifier 35 for the current-voltage that the signal that mercury-cadmium tellurid detector 34 exports first passes around a DC-100MHz amplifies, Ran Houjing
The filtering for crossing 0~500MHz low pass filters 36 is input in the capture card (not shown) of NI5152 types, is used afterwards
Labview data acquisition softwares, which are realized the data acquisition of nanosecond-second-time and entered, carries out data preservation in computer.
To sum up, mid-infrared light spectrometer is differentiated using the nanosecond pulse heating-up time provided by the present invention to have the following effects that：
1) wide mid-infrared light spectrometry can be realized, the absorption of secondary protein structure is completely covered；2) can obtain high-precision
Transient state ir data, measurement accuracy can reach 1 × 10-4OD；3) data acquisition of the nanosecond to second-time can be realized.
The running of mid-infrared light spectrometer is described in detail with reference to the measurement to Cyt C protein transient state infrared spectrum
Journey.
1) preparation of samples：The double sample sample cells in a pond are sampled, and heavy water is loaded onto as reference, second area in its first area
The heavy aqueous solution dissolved with 1% cytochrome c protein is loaded onto as sample.
2) experiment condition：25 DEG C of basal temperature, Ho laser single-pulse energies 17mj, warm skip frame degree is about 11 DEG C, detects light
Spot diameter is less than 100 microns, and pumping optical beam spot diameter is about 1mm.
3) by the wavelength regulation of tunable CO lasers to the wavelength for wanting measurement：Such as 1645cm-1At wave number, respectively
Nanosecond pulse heating experiment is carried out to reference and sample.Now, as pulse heats up to the moment of heavy water, through reference and sample
1645cm-1Laser energy can change, call the collection of NI data collecting cards to pass through reference using labview capture programs
Changed with time with the laser energy of sample, and be converted into absorption intensity (OD) value.The reference and the signal of sample collected
As shown in figures 5-9.
Wherein, Fig. 5 is heavy water in 1645cm-1The measurement result of the Transient Dynamics signal at place twice.Fig. 6 is heavy water and sample
Product are in 1645cm-1In signal schematic representation, as can be seen from Figure 6 the signal of sample be greater than the signal of reference.Fig. 7 is twice
The difference of the dynamic signal of heavy water, it can be seen from figure 7 that the signal to noise ratio of the mid-infrared light spectrometer of the present invention has reached 10- 4OD precision.
4) sample signal of the obtained protein in heavy water is subtracted into the signal of heavy water with regard to that can obtain 1645cm-1That locates is thin
Change kinetics information of the Cytochrome C albumen after the rise of temperature moment, as shown in Figure 8.As can be seen from Figure 8 temperature wink
Between when raising, cromoci is in 1645cm-1The absorbance at place reduces, and reaches stable after stabilizing the temperature.
5) and then the output wavelengths of tunable CO lasers, repeat step 3 are changed) and 4) with regard to that can obtain at different wave numbers
Protein dynamics information.
6) the same time such as 2 after the dynamic information at the different wave numbers for obtaining step 5) takes heating pulse next
Value at microsecond is mapped (for example, 1645cm-1Locate as 2.5 × 10-3OD), it is possible to it is thin to obtain 2 microseconds after heating pulse reaches
The transient state infrared spectrum of Cytochrome C albumen, as shown in Figure 9.As can be seen from Figure 9 after moment heating, cromoci exists
1620cm-1-1645cm-1The absorbance at place reduces, in 1662cm-1And 1683cm-1The absorbance at place rises, it is meant that cell color
Plain C is after moment heats up, hydrophilic helical structure (1620cm-1-1645cm-1) disappearance and disordered structure (1662cm-1)
With corner (1683cm-1) generation.
So far, although those skilled in the art will appreciate that detailed herein have shown and described multiple showing for the present invention
Example property embodiment, still, without departing from the spirit and scope of the present invention, still can be straight according to present disclosure
Connect determination or derive many other variations or modifications for meeting the principle of the invention.Therefore, the scope of the present invention be understood that and
Regard as covering other all these variations or modifications.
Claims (9)
1. a kind of mid-infrared light spectrometer, the dynamic structure for measuring protein changes, and the mid-infrared light spectrometer includes：
Wideband is tunable CO laser (10), it is described for sending middle infrared acquisition laser to testing protein solution
Wideband is tunable, and CO laser (10) uses energisation mode of the bidirectional high-pressure electric pump as laser, high pressure electric pump
Concrete structure is：Laser cavity both ends are anodes, and two anodes share middle negative electrode, and one end of laser cavity is gold-plated concave mirror,
The other end is the grating that Electronic control rotates, and the gold-plated concave mirror is on fine-tuning mirror holder, to enter to the laser cavity
Row fine setting, the grating is vertically arranged with a plane gold mirror, and is fixed on same turntable, to export when grating rotates
The direction of wavelength change but output laser does not change；
Nanosecond adjusts Q holmium lasers (20), and for sending pumping laser to heat the testing protein solution, the nanosecond adjusts Q
Holmium laser (20) laser cavity uses flat cavity configuration, and one end is total reflective mirror (21), and one end is the output coupling that delivery efficiency is 20%
Close mirror (22), nanosecond short pulse adjust Q holmium lasers to use doped with 1.5%Cr, 5.8%Tm, Cr, the Tm of 0.35%Ho elements,
Ho:YAG crystal is used as pumping source, electro-optic Q-switched crystal La as laser crystal (23), flash lamp (24)3Ga5SiO14(LGS)
Crystal (25), Q mode of operations are adjusted using pressure type is moved back, the polarizer is a pair of alpha-alumina crystals (26) that Brewster's angle is placed, and is used
λ/4 quartz wave-plate (27) modulates Depolarization；
Pump probe light path system, for detecting through the strong of the middle infrared acquisition laser after the testing protein solution
Degree；And
Data collecting system (40), for gathering the data of the intensity detected from the pump probe light path system；
For placing the sample cell (50) of the testing protein solution, the sample cell (50), which has, to be used to place reference weight
The first area (51) of water and the second area (52) for placing the heavy aqueous solution containing the testing protein solution；
The pumping laser for adjusting Q holmium lasers (20) to send the nanosecond forms the spot diameter on the sample cell (50)
For 2mm.
2. mid-infrared light spectrometer according to claim 1, it is characterised in that the pump probe light path system is included successively
Light path part, detector, amplifier and the wave filter of setting.
3. mid-infrared light spectrometer according to claim 2, it is characterised in that the detector is liquid nitrogen refrigerating type high speed tellurium
Cadmium mercury detector；The amplifier is DC-100MHz amplifiers.
4. the mid-infrared light spectrometer according to any one of claim 1-3, it is characterised in that the tunable oxygen of wideband
The output wavelength for changing carbon laser is 5.2 μm~6.2 μm.
5. mid-infrared light spectrometer according to claim 4, it is characterised in that the nanosecond adjusts the output arteries and veins of Q holmium lasers
A width of 50ns, output wavelength 2080nm.
6. mid-infrared light spectrometer according to claim 1, it is characterised in that the sample cell (50) is rotary structure,
And the wideband tunable one can be received by its described first area (51) of rotation and the second area (52)
The middle infrared acquisition laser and the nanosecond that carbon oxide laser device (10) is sent adjust the pump that Q holmium lasers (20) are sent
Pu laser.
7. mid-infrared light spectrometer according to claim 6, it is characterised in that
The middle infrared acquisition laser that the tunable CO laser of wideband (10) sends is formed in the sample cell
(50) spot diameter on is 0.4mm.
8. the mid-infrared light spectrometer according to any one of claim 1-3,5-7, it is characterised in that the data acquisition system
System (40) includes 8 high-speed collection cards, and minimum is distinguished as 0.36mV.
9. mid-infrared light spectrometer according to claim 8, it is characterised in that the data collecting system (40) be arranged to
The down-sampled type collection data of nanosecond to second time frame.
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| Title |
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| Dynamics of thermal microeffects rate constants of thermal denaturation measured by a temperature-jump experiment;Gereon Huttmann，Reginald Birngruber;《Biomedical Optical Spectroscopy and Diagnostics/Therapeutic Laser Applications》;19981231;第300-305页 * |
| 蛋白质快速折叠动力学的脉冲升温时间分辨时间红外光谱研究;叶满萍;《中国科学院物理研究所博士学位论文》;20081231;第13-38页 * |
| 高稳定性宽频带一氧化碳激光器的研究;陈潇为;《中国优秀硕士学位论文全文数据库 信息科技辑》;20110915;第4-5、45页 * |
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