CN108152252B - Integral femtosecond time resolution fluorescence life measuring spectrometer - Google Patents
Integral femtosecond time resolution fluorescence life measuring spectrometer Download PDFInfo
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- 238000001514 detection method Methods 0.000 claims abstract description 31
- 230000003287 optical effect Effects 0.000 claims abstract description 22
- 238000001228 spectrum Methods 0.000 claims abstract description 21
- 238000005086 pumping Methods 0.000 claims abstract description 14
- 239000000523 sample Substances 0.000 claims description 48
- 238000005259 measurement Methods 0.000 claims description 18
- 230000010354 integration Effects 0.000 claims description 9
- 239000013307 optical fiber Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 2
- 238000002189 fluorescence spectrum Methods 0.000 abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 6
- WISWLZYHZLVSMO-UHFFFAOYSA-N 6-phenyl-2-(4-phenylphenyl)-1,3-benzoxazole Chemical compound C1=CC=CC=C1C1=CC=C(C=2OC3=CC(=CC=C3N=2)C=2C=CC=CC=2)C=C1 WISWLZYHZLVSMO-UHFFFAOYSA-N 0.000 description 4
- 230000005855 radiation Effects 0.000 description 4
- 230000003595 spectral effect Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005316 response function Methods 0.000 description 2
- 229910000737 Duralumin Inorganic materials 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 238000013032 photocatalytic reaction Methods 0.000 description 1
- 238000006552 photochemical reaction Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 238000000215 ultra-fast laser spectroscopy Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6408—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence
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Abstract
The invention relates to a time-resolved fluorescence spectrum measuring instrument, in particular to an integral femtosecond time-resolved fluorescence lifetime measuring spectrometer which comprises a detection and pumping laser optical path system, a sample fluorescence acquisition system and a spectrum detection system. The spectrometer excites a sample by using one beam of femtosecond laser, the other beam of femtosecond laser enables the excited sample to be excited and radiated, and all collected sample fluorescence is integrated in a spectrum detection system, so that the fluorescence life is measured.
Description
Technical Field
The invention relates to a time-resolved spectral measuring instrument, in particular to an integral femtosecond time-resolved fluorescence lifetime measuring spectrometer, which excites a sample by using one beam of femtosecond laser, then excites the excited sample to be radiated by using the other beam of femtosecond laser, so that the fluorescence quantity of free radiation of the sample is reduced, a spectral detection system integrates all collected sample fluorescence, compares the fluorescence integral intensity when the excited radiation exists and the fluorescence integral intensity when the excited radiation does not exist, obtains a fluorescence lifetime signal point, and realizes the measurement of the femtosecond time-resolved fluorescence lifetime by matching with a time delay technology.
Background
The time-resolved pumping-detection technology [ ultrafast laser spectroscopy principle and technology basis, compiled by the Wengxiang and Chenhailong, 2013, the chemical industry publisher ], also known as excitation-detection double-pulse technology [ time-resolved spectroscopy basis, Guo basis, 2012, advanced education publisher ], has wide application in various fields of modern science, such as photophysical process, photochemical reaction, biochemical process, photocatalytic reaction, energy and charge transmission process, nano material characterization and the like. The fluorescence lifetime measurement has many applications in the aspects of biological fluorescence probe analysis, solar energy utilization, fluorescent molecule photophysical photochemical property, substance excited state dynamics research and the like; the femtosecond time resolution spectrum instrument related by the invention also applies the pumping-detection technology. Korean clely et al [ chinese patent application No.: 201310392018.6, a femtosecond time-resolved transient absorption and fluorescence depletion two-in-one spectrometer is built, two pumping-detecting technologies are combined into one spectrometer, and the application of the femtosecond time-resolved spectrometer is expanded.
The current commonly used fluorescence life measuring instruments mainly have time-related single photon counters, stripe cameras, femtosecond time resolution fluorescence up-conversion spectrometers, femtosecond time resolution fluorescence depletion spectrometers and the like. The time resolution of the time correlation single photon counter is also dozens of picoseconds at most; the time resolution of the stripe camera can reach picosecond level at most, but the stripe camera is very expensive; the time resolution of the femtosecond time resolution fluorescence upconversion spectrometer is limited by the upconversion crystal and is generally two times larger than the pulse width of the used femtosecond laser; the femtosecond time resolution fluorescence depletion spectrum instrument has high time resolution, but only one fluorescence wavelength is usually measured when the femtosecond time resolution fluorescence depletion spectrum instrument is used for measuring fluorescence, so that the application of the femtosecond time resolution fluorescence depletion spectrum instrument in the aspect of measuring the fluorescence life of a weak fluorescent substance is reduced.
Disclosure of Invention
The invention aims to obtain an integral femtosecond time-resolved fluorescence lifetime measurement spectrometer which has good time resolution and fluorescence intensity detection limit.
In order to achieve the purpose, the invention adopts the technical scheme that:
an integral femtosecond time-resolved fluorescence life measurement spectrometer comprises a detection and pumping laser light path system, a sample fluorescence acquisition system and a spectrum detection system;
all the components are arranged on the optical flat plate;
the femtosecond laser detection beam and the femtosecond laser pumping beam are overlapped in a space intersection manner in the sample pool;
the femtosecond laser probe beam and the femtosecond laser pumping beam are crossed in space and have a certain time interval, and the time interval is adjusted by a time delayer;
the femtosecond laser detection beam is adjusted by a time delayer, passes through a delayer rear reflector and a sample cell front reflector, and is spatially crossed and superposed with the femtosecond laser pumping beam on the sample cell;
the sample fluorescence collection system is characterized in that fluorescence radiated by a sample is collected by a fluorescence collection mirror, reflected to a fluorescence focusing mirror by a fluorescence reflecting mirror, focused to an optical fiber inlet by the fluorescence focusing mirror and then guided into a spectrometer by an optical fiber;
the spectrum measurement system is that after the fluorescence is split by the spectrometer, a polychromatic spectrum is formed on the detector, and the polychromatic spectrum on the detector is integrated by a computer to obtain a fluorescence integral signal;
after the optical chopper operates, two fluorescence integral signals of a femtosecond-free laser detection beam and a femtosecond laser detection beam are obtained on a computer, and the difference value of the two fluorescence integral signals is a fluorescence life signal point;
and drawing a curve of the fluorescence lifetime signal point to the delay time on a computer to obtain an integral femtosecond time-resolved fluorescence lifetime curve of the sample.
The optical flat plate material is duralumin or stainless steel, and the size specification is as follows: a length of about 120cm, a width of about 60cm and a thickness of about 1 cm;
the fluorescence collecting mirror and the fluorescence focusing mirror can be but are not limited to a lens, a paraboloidal mirror and a spherical mirror;
the sample cell is a cuvette with an optical path of 1mm or 2mm or 10 mm;
the trigger frequency signal of the optical chopper is derived from a femtosecond laser, and the chopping frequency of the optical chopper is set between 20Hz and 1000 Hz;
the detector may be, but is not limited to, a photodiode array, a charge coupled device.
Drawings
FIG. 1 is a schematic view of the structure of an integrated femtosecond time-resolved fluorescence lifetime measurement spectrometer according to the present invention.
FIG. 2 is an integrated femtosecond time-resolved fluorescence lifetime measurement curve of PBBO dye in ethanol solution with a concentration of 1mmol/L in example 1.
FIG. 3 is an integrated femtosecond time-resolved fluorescence lifetime measurement curve of 0.1mmol/L OX750 dye in ethanol solution in example 2. (a) A spectral evolution curve of the fluorescence spectrum of the OX750 solution along with the change of time; (b) the curve of the fluorescence intensity at 660nm as a function of the delay time (filled box) and the curve of the total fluorescence intensity after integration of the fluorescence intensities at all wavelengths as a function of the delay time (empty box).
Detailed Description
Referring to the drawings, fig. 1 is a schematic structural diagram of the present invention. The integrated femtosecond time-resolved fluorescence life measuring spectrometer is characterized in that an optical flat plate is about 120cm long and 60cm wide and is provided with two light inlets, wherein a first light inlet 2 is a femtosecond laser detection beam inlet and is used for stimulated radiation of a sample; the second light inlet 3 is a femtosecond laser pumping beam inlet and is used for exciting a sample.
All components are arranged on an optical flat plate 1, the femtosecond laser detection beam 2 and the femtosecond laser pumping beam 3 are spatially crossed and superposed, the femtosecond laser detection beam 2 is adjusted by a time delayer 5 and then passes through a delayer rear reflector 6 and a sample pool front reflector 7, and the sample in the sample pool 4 is spatially crossed and superposed with the femtosecond laser pumping beam 3;
the femtosecond laser probe beam 2 and the femtosecond laser pump beam 3 are crossed and superposed in space at the position of a sample in the sample pool 4 and have a specific time interval, and the time interval is adjusted by a time delayer 5;
the sample fluorescence collection system is characterized in that fluorescence radiated by a sample is collected by a fluorescence collection mirror 8, reflected to a fluorescence focusing mirror 10 by a fluorescence reflection mirror 9, focused to an inlet of an optical fiber 11 by the fluorescence focusing mirror 10, and then guided to a spectrometer 12 through the optical fiber;
in the spectrum measurement system, after fluorescence is split by the spectrometer 12, a polychromatic spectrum is formed on the detector 13, and the polychromatic spectrum on the detector 13 is subjected to integration processing by the computer 14 to obtain a fluorescence integration signal;
after the optical chopper 15 operates, two fluorescence integral signals of no femtosecond laser detection beam and the femtosecond laser detection beam are obtained on a computer, and the difference value of the two fluorescence integral signals is a fluorescence life signal point;
the fluorescence lifetime signal points are plotted into a curve for the delay time on the computer 14 and fitted to obtain the integral femtosecond time-resolved fluorescence lifetime of the sample.
The fluorescence collection mirror 8 and the fluorescence focusing mirror 10 may be, but are not limited to, a lens, a parabolic mirror, or a spherical mirror;
the sample cell 4 is a cuvette with an optical path of 1mm or 2mm or 10 mm;
an optical chopper 15 whose trigger frequency signal is derived from a femtosecond laser, and whose chopping frequency is set between 20Hz and 1000 Hz;
the detector 13 may be, but is not limited to, a photodiode array or a charge coupled element.
Example 1
Aiming at the time resolution capability of the integrated femtosecond time resolution fluorescence life measuring spectrometer in the performance of measuring the integrated femtosecond time resolution fluorescence life, the integrated femtosecond time resolution fluorescence life signal is measured by adopting 1m mol/L PBBO dye ethanol solution as a test sample. The peak of the absorption spectrum of PBBO is around 320nm, and the fluorescence of PBBO can be excited only when two light pulses of 490nm pumping beam and 800nm detecting beam coincide at the sample in the sample cell in space and time, so that the time-resolved response function of the integrated femtosecond time-resolved fluorescence lifetime measurement spectrometer can be obtained. The instrument response function obtained in this example was fitted using a gaussian function, resulting in a standard deviation of 0.104 picosecond and a full width at half maximum of 0.246 picosecond. The experimental data is shown in figure 2.
Example 2
Aiming at the investigation of the fluorescence intensity detection limit performance of the integral femtosecond time-resolved fluorescence lifetime measurement spectrometer, 0.1m mol/L ethanol solution of OX750 dye is used as a sample; the pump light is 490nm, 1.5 mW; the detection light is 800nm and 1 mW. The integrated femtosecond time-resolved fluorescence lifetime measurement spectrum of the measurement sample has experimental data shown in figure 3. Wherein the left panel (a) is the evolution of the measured OX750 fluorescence spectrum as a function of delay time; the right panel (b) includes the decay curve of fluorescence intensity with delay time at a single wavelength of 660nm without integration of the OX750 fluorescence peak (filled boxes), and the decay curve of integrated fluorescence intensity with delay time obtained after integration of the OX750 fluorescence peak (empty boxes). It can be seen that the noise of the curve after integration is significantly less than before integration. This is a curve showing the generation of OX750 fluorescence signal.
Claims (4)
1. An integral femtosecond time-resolved fluorescence life measurement spectrometer comprises a detection and pumping laser light path system, a sample fluorescence acquisition system and a spectrum detection system; the method is characterized in that:
the detection and pumping laser optical path system comprises a sample cell, a time delayer (5) and an optical chopper (15), and the sample fluorescence acquisition system comprises a spectrometer (12), a detector (13) and a computer (14); the femtosecond laser probe beam (2) emitted by the femtosecond laser irradiates a sample in the sample cell (4) through the time delayer (5), the optical chopper (15) and the femtosecond laser pump beam (3) emitted by the femtosecond pump laser, and the femtosecond laser pump beam (3) enables the sample to generate fluorescence; after sample fluorescence is split by a light splitting spectrometer (12), a multicolor spectrum is formed on a detector (13), and the multicolor spectrum on the detector (13) is integrated by a computer (14) to obtain a fluorescence integration signal;
when the optical chopper (15) is operated, integral signals of two fluorescence intensities to wavelengths of a femtosecond-free laser detection beam and a femtosecond laser detection beam are obtained on a computer (14), and the difference value of the two fluorescence integral signals is a fluorescence life signal point; drawing a curve on a computer (14) by taking the delay time as an abscissa and the fluorescence lifetime signal point as an ordinate to obtain an integral femtosecond time-resolved fluorescence lifetime measurement curve of the sample;
the sample fluorescence collection system is characterized in that fluorescence radiated by a sample is collected by a fluorescence collection mirror (8), reflected to a fluorescence focusing mirror (10) by a fluorescence reflection mirror (9), focused to an optical fiber (11) by the fluorescence focusing mirror (10), and then enters a spectrometer (12) from the optical fiber (11); the fluorescence collecting mirror (8) and the fluorescence focusing mirror (10) can be but are not limited to a lens, a paraboloidal mirror or a spherical mirror respectively;
the sample cell (4) is a cuvette with an optical path of 1mm, 2mm or 10 mm; the femtosecond laser detection beam (2) and the femtosecond laser pumping beam (3) are in cross coincidence in the space where the sample is located in the sample pool, and the femtosecond laser detection beam (2) is adjusted by a time delayer (5) and an optical chopper (15), passes through a retarder rear reflector (6) and a sample pool front reflector (7), and is in cross coincidence with the femtosecond laser pumping beam (3) in the space on the sample pool (4);
the spectrum detection system is characterized in that after fluorescence is split by the spectrometer (12), a polychromatic spectrum is formed on the detector (13), and the polychromatic spectrum on the detector (13) is integrated by the computer (14) to obtain a fluorescence integration signal; the detector (13) may be, but is not limited to, a photodiode array or a charge coupled element.
2. The integrated femtosecond time-resolved fluorescence lifetime measurement spectrometer according to claim 1, wherein:
all components in the detection and pumping laser optical path system, the sample fluorescence acquisition system and the spectrum detection system are arranged on the optical flat plate (1).
3. The integrated femtosecond time-resolved fluorescence lifetime measurement spectrometer according to claim 1, wherein:
the trigger frequency of the optical chopper (15) is derived from a femtosecond laser, and the chopping frequency of the optical chopper is set between 20Hz and 1000 Hz.
4. The integrated femtosecond time-resolved fluorescence lifetime measurement spectrometer according to claim 1 or 3, wherein:
the computer (14) is connected with the time delayer (5), the femtosecond laser and the detector (13) through data lines.
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