CN109991197A - Measure the spectrophotometer and method of fluorescent characteristic - Google Patents
Measure the spectrophotometer and method of fluorescent characteristic Download PDFInfo
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- CN109991197A CN109991197A CN201711486648.4A CN201711486648A CN109991197A CN 109991197 A CN109991197 A CN 109991197A CN 201711486648 A CN201711486648 A CN 201711486648A CN 109991197 A CN109991197 A CN 109991197A
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- 238000000034 method Methods 0.000 title claims abstract description 9
- 238000001228 spectrum Methods 0.000 claims abstract description 38
- 230000003287 optical effect Effects 0.000 claims abstract description 19
- 238000002189 fluorescence spectrum Methods 0.000 claims abstract description 12
- 238000012360 testing method Methods 0.000 claims abstract description 12
- 238000005259 measurement Methods 0.000 claims description 12
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 claims description 3
- 229910052805 deuterium Inorganic materials 0.000 claims description 3
- 238000004611 spectroscopical analysis Methods 0.000 claims description 3
- WFKWXMTUELFFGS-UHFFFAOYSA-N tungsten Chemical compound [W] WFKWXMTUELFFGS-UHFFFAOYSA-N 0.000 claims description 3
- 229910052721 tungsten Inorganic materials 0.000 claims description 3
- 239000010937 tungsten Substances 0.000 claims description 3
- 230000003321 amplification Effects 0.000 abstract description 3
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 3
- 230000005540 biological transmission Effects 0.000 description 5
- 238000000862 absorption spectrum Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000000985 reflectance spectrum Methods 0.000 description 2
- 238000009738 saturating Methods 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000000411 transmission spectrum Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/44—Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
- G01J3/4406—Fluorescence spectrometry
-
- 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
-
- 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
- G01N2021/6417—Spectrofluorimetric devices
-
- 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
- G01N2021/6491—Measuring fluorescence and transmission; Correcting inner filter effect
Abstract
The present invention relates to a kind of spectrophotometers and method for measuring fluorescent characteristic, characterized in that includes: light source, for issuing required spectrum；The first sample room is all incident on for placing sample to be tested, with the unselected light that the light source issues in first sample room, first sample room；Monochromator, monochromator is by the complex light of light source or is decomposed into monochromatic light through the complex light of the first sample room and can arbitrarily change wavelength；Second sample room is incident to the second sample room for placing sample to be tested with the light of whole wavelength of monochromator selection；Detector, the optical signal for will transmit through sample to be tested is converted into electric signal, and electric signal is amplified；And display and storage system, the electric signal for will test device amplification is shown by instrument, and stores optical signal.The present invention solves the problems, such as that fluorescence measures transmitted spectrum and generates interference, while obtaining the transmitted spectrum and fluorescence spectrum of sample to be tested, and improves quantitative accuracy.
Description
Technical field
The present invention relates to a kind of spectrophotometer, especially a kind of spectrophotometer for measuring fluorescent characteristic and side
Method.
Background technique
Spectrophotometer is using spectrophotometry, by measurement sample in certain wave strong point or a wavelength range
The trap or reflectance of interior light carry out qualitative and quantitative analysis to sample.Referring to Fig.1, spectrophotometer mainly by light source,
Monochromator, sample room, detector and display are formed with storage system.Spectrophotometer generallys use double light path photometry at present
Framework is measured, as shown in Fig. 2, mainly including light source 1, monochromator 2, diaphragm 3, polarizing film 4, beam splitter 5, attenuator 6, focusing is thoroughly
Mirror 7, with reference to optical detector 8, specimen holder 9, sample 10, condenser lens 11, test optical detector 12.What secondary color light source 1 was emitted answers
Coloured light forms monochromatic light required for measurement after monochromator 2, and monochromatic light forms measurement institute after diaphragm 3 and polarizing film 4
The linearly polarized light needed, linearly polarized light form a branch of measurement light and a branch of reference light after beam splitter 5, and reference light passes through attenuator
6 and condenser lens 7 after be referenced optical detector 8 collection, test light after sample 10 and condenser lens 11 be tested optical detection
Device is received.Existing spectrophotometer, the test method and principle of sample transmission rate are as follows: first in measuring beam and reference light
Shu Zhongjun does not place sample room, acquires the light of reference beam and test beams simultaneously with reference optical detector and test optical detector
By force, it is denoted as I respectively1And I2, then sample to be tested is placed in optical system for testing, again using with reference to optical detector and test light
Detector acquires the light intensity of reference beam and test beams simultaneously, is denoted as I respectively1 *And I2 *, then the transmissivity T of sample to be tested can
It is calculated according to formula (1.1):
I2 *×I1/I1 *×I2(1.1)；
The wavelength X of incident light is gradually changed using monochromator, successively measures the transmissivity of sample at different wavelengths
Obtain the transmitted spectrum T (λ) of sample to be tested.
And for fluorescent material, the incident light irradiation through certain wavelength enters excitation state after absorbing luminous energy, and stands
I.e. de excitation is sent out and issues the emergent light longer than the wavelength of incident light, as fluorescence or phosphorescence.If with existing spectrophotometric
Meter and measurement method, measure its a certain wave band transmission perhaps reflectance spectrum because fluorescence or phosphorescence influence, it is measured
Transmission or reflectance spectrum in fluorescence area have biggish error.Therefore a new spectrophotometer apparatus is needed to differentiate
With eliminate this error, while measuring the fluorescence spectrum of sample.
Summary of the invention
The purpose of the present invention is overcoming the deficiencies in the prior art, a kind of light splitting for measuring fluorescent characteristic is provided
Photometer and method solve the problems, such as that fluorescence or phosphorescence measure transmitted spectrum and generate interference, while obtaining sample to be tested
Transmitted spectrum and fluorescence spectrum, and improve quantitative accuracy.
According to technical solution provided by the invention, a kind of spectrophotometer measuring fluorescent characteristic, characterized in that packet
It includes:
Light source, for issuing required spectrum；
First sample room, first sample room are issued unselected for placing sample to be tested with the light source
Light is all incident on the first sample room；
The complex light of the first sample room of the complex light of light source or transmission is decomposed into monochromatic light and can by monochromator, monochromator
Arbitrarily change wavelength；
Second sample room is incident to second for placing sample to be tested with the light of whole wavelength of monochromator selection
Sample room；
Detector, the optical signal for will transmit through sample to be tested is converted into electric signal, and electric signal is amplified；
And display and storage system, the electric signal for will test device amplification is shown by instrument, and stores light
Signal；
The light source, the first sample room, monochromator, the second sample room, detector and display and storage system are suitable according to optical path
Secondary connection.
Further, the light source is tungsten lamp or deuterium lamp.
Further, the monochromator is prism or grating.
Further, the detector is photoelectric tube or photomultiplier tube.
The present invention also provides a kind of methods for measuring fluorescent characteristic, characterized in that the following steps are included:
(1) it does not place sample in the first sample room, the second sample room first, carries out baseline correction；
(2) after the completion of baseline correction, sample to be tested is placed in the first sample room, the light that light source issues is not chosen complete
Portion is incident in the sample to be tested of the first sample room, and the light of subband is absorbed；Through the saturating of the first sample room sample to be tested
It penetrates fluorescence that light and sample to be tested may issue or phosphorescence mixes, resolve into monochromatic light exposure to detection through monochromator
On device, the first transmitted spectrum of sample to be tested in the first sample room is obtained；
(3) sample to be tested is placed in the second sample room, the full spectrum light that light source issues is by becoming single after monochromator
Coloured light, monochromatic light are incident in the sample to be tested of the second sample room, and the monochromatic light of some wave bands is absorbed；Through the second sample room
It is irradiated on detector after fluorescence or the phosphorescence superposition that the monochromatic light and sample to be tested of sample to be tested may issue, obtains second
Second transmitted spectrum of sample to be tested in sample room.
(4) spectroscopic data of the first transmitted spectrum, the second transmitted spectrum is analyzed, the two is identical, illustrates that sample to be tested does not have
There is fluorescent characteristic；If the two data have differences in a certain wave band, illustrate that sample to be tested has fluorescent characteristic；First transmission
Difference between spectrum and the second transmitted spectrum is the fluorescence spectrum for being approximately sample to be tested.
Beneficial effects of the present invention:
The present invention provides a kind of spectrophotometers for being capable of measuring fluorescent characteristic, eliminate fluorescence or phosphorescence to saturating
The interference of spectral measurement is penetrated, while obtaining the transmitted spectrum and fluorescence spectrum of sample to be tested, and improves measurement sensitivity and determines
Measure accuracy.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of existing spectrophotometer.
Fig. 2 is the optical path schematic diagram of existing spectrophotometer.
Fig. 3 is the structural schematic diagram of spectrophotometer of the invention.
Fig. 4 is the transmitted spectrum and fluorescence spectrum of spectrophotometer measurement of the invention.
Specific embodiment
Below with reference to specific attached drawing, the invention will be further described.
As shown in figure 3, the spectrophotometer of measurement fluorescent characteristic of the present invention, including it is sequentially connected according to optical path
Light source 20, the first sample room 21, monochromator 22, the second sample room 23, detector 24 and display and storage system 25.
The light source 20 can be tungsten lamp or deuterium lamp for issuing required spectrum；
First sample room 21 is all entered for placing sample to be tested with the unselected light that the light source issues
It is mapped to the first sample room 21；
The monochromator 22, for being decomposed into monochrome by the complex light of light source 20 or through the complex light of the first sample room 21
Light simultaneously can arbitrarily change wavelength, can be prism or grating；
Second sample room 23 is entered for placing sample to be tested with the light of whole wavelength of the monochromator 22 selection
It is incident upon the second sample room 23；
The detector 24, the optical signal for will transmit through sample to be tested is converted into electric signal, and electric signal is amplified, can
For photoelectric tube or photomultiplier tube；
The display and storage system 25, the electric signal for will test the amplification of device 24 is shown by instrument, and is deposited
Store up optical signal.
The present invention measures fluorescent material transmitted spectrum and the method for fluorescence spectrum is:
(1) it does not place sample in the first sample room 21, the second sample room 23 first, carries out baseline correction；
(2) after the completion of baseline correction, sample to be tested is placed in the first sample room 21, measures the first transmitted spectrum A；
(3) sample to be tested is placed in the second sample room 23, measures the second transmitted spectrum B；
(4) data are analyzed, and the first transmitted spectrum A and the second transmitted spectrum B are compared, as shown in figure 4, judging sample to be tested
Whether there is fluorescent characteristic, and measures the fluorescence spectrum of sample to be tested.
The working principle of the spectrophotometer of measurement fluorescent characteristic of the present invention:
Firstly, sample to be tested is placed in the first sample room 21, the not chosen whole of light that light source 20 issues is incident on
In sample to be tested, the light of subband is absorbed, if sample to be tested has fluorescent characteristic, is absorbed the light of certain wavelength, will be sent out
The fluorescence or phosphorescence longer than the wavelength of incident light out, the fluorescence that transmitted light and sample to be tested through sample to be tested may issue
Or phosphorescence mixes, and resolves on monochromatic light exposure to detector 24 through monochromator 22, is gradually changed with monochromator 22
The wavelength of transmitted light successively measures the transmissivity of sample to be tested different wave length, obtains the first transmitted spectrum A of sample to be tested；
Secondly, sample to be tested is placed in the second sample room 23, the full spectrum light that light source 20 issues passes through monochromator 22
After become monochromatic light, the monochromatic light of seleced specific wavelength is incident in sample to be tested, according to sample absorption Spectrum characteristic, one
The monochromatic light of a little wave bands is absorbed, if sample to be tested has fluorescent characteristic, absorbs monochromatic light, will be issued more monochromatic than incident
The long fluorescence of wavelength fluorescence or phosphorescence that perhaps phosphorescence may be issued through the monochromatic light and sample to be tested of sample to be tested are superimposed
After be irradiated on detector, the sum of light intensity of the two is by as the monochromatic transmitted light intensity of this wavelength.Gradually changed using monochromator
Become the wavelength of incident light, successively measures the transmissivity of sample to be tested at different wavelengths, obtain the second transmitted light of sample to be tested
Compose B.
Data analysis: the spectroscopic data of the first transmitted spectrum A of analysis, the second transmitted spectrum B, the two is identical, illustrates to be measured
Sample does not have fluorescent characteristic；If the two data have differences in a certain wave band, illustrate that sample to be tested has fluorescent characteristic.
First transmitted spectrum A is that sample to be tested own absorption spectrum and being stimulated under full spectral illumination generates the folded of fluorescence spectrum
Add, the absorption spectrum for showing as sample has been superimposed fluorescence spectrum；A certain monochromatic transmitance is to be measured in second transmitted spectrum B
Sample monochromatic absorption and the combined influence in this monochromatic light institute exciting light herein, show as the absorption spectrum of sample because
Test absorbance is resulted in by the compensation of sample excitation light and is less than real absorbance；If the fluorescence quantum efficiency of sample is enough
Small, the difference between the first transmitted spectrum A and the second transmitted spectrum B is the fluorescence spectrum for being approximately sample to be tested.
Claims (5)
1. a kind of spectrophotometer for measuring fluorescent characteristic, characterized in that include:
Light source (20), for issuing required spectrum；
First sample room (21), first sample room (21) are issued not for placing sample to be tested with the light source (20)
Seleced light is all incident on the first sample room (21)；
Monochromator (22), monochromator (22) are decomposed by the complex light of light source (20) or through the complex light of the first sample room (21)
Monochromatic light simultaneously can arbitrarily change wavelength；
Second sample room (23) is incident to for placing sample to be tested with the light of whole wavelength of the monochromator (22) selection
Second sample room (23)；
Detector (24), the optical signal for will transmit through sample to be tested is converted into electric signal, and electric signal is amplified；
And show with storage system (25), it is shown for will test the electric signal that device (24) amplify by instrument, and deposit
Store up optical signal；
The light source (20), the first sample room (21), monochromator (22), the second sample room (23), detector (24) and display with
Storage system (25) is sequentially connected with according to optical path.
2. the spectrophotometer of measurement fluorescent characteristic as described in claim 1, it is characterized in that: the light source (20) is tungsten
Lamp or deuterium lamp.
3. the spectrophotometer of measurement fluorescent characteristic as described in claim 1, it is characterized in that: the monochromator (22) is
Prism or grating.
4. the spectrophotometer of measurement fluorescent characteristic as described in claim 1, it is characterized in that: the detector (24) is
Photoelectric tube or photomultiplier tube.
5. a kind of method for measuring fluorescent characteristic, characterized in that the following steps are included:
(1) it does not place sample in the first sample room (21), the second sample room (23) first, carries out baseline correction；
(2) after the completion of baseline correction, sample to be tested is placed in the first sample room (21), the light that light source (20) issues is without choosing
It selects and is all incident in the sample to be tested of the first sample room (21), the light of subband is absorbed；Through the first sample room (21)
The fluorescence or phosphorescence that the transmitted light and sample to be tested of sample to be tested may issue mix, and resolve into through monochromator (22)
On monochromatic light exposure to detector (24), the first transmitted spectrum (A) of sample to be tested in the first sample room (21) is obtained；
(3) sample to be tested is placed in the second sample room (23), the full spectrum light that light source (20) issues passes through monochromator (22)
After become monochromatic light, monochromatic light is incident in the sample to be tested of the second sample room (23), and the monochromatic light of some wave bands is absorbed；Thoroughly
It is irradiated to after fluorescence or the phosphorescence superposition that the monochromatic light and sample to be tested for crossing the second sample room (23) sample to be tested may issue
On detector (24), the second transmitted spectrum (B) of sample to be tested in the second sample room (23) is obtained.
(4) spectroscopic data of the first transmitted spectrum (A), the second transmitted spectrum (B) is analyzed, the two is identical, illustrates sample to be tested not
With fluorescent characteristic；If the two data have differences in a certain wave band, illustrate that sample to be tested has fluorescent characteristic；First thoroughly
The difference penetrated between spectrum (A) and the second transmitted spectrum (B) is approximately the fluorescence spectrum of sample to be tested.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201711486648.4A CN109991197A (en) | 2017-12-29 | 2017-12-29 | Measure the spectrophotometer and method of fluorescent characteristic |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711486648.4A CN109991197A (en) | 2017-12-29 | 2017-12-29 | Measure the spectrophotometer and method of fluorescent characteristic |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111829971A (en) * | 2020-08-10 | 2020-10-27 | 广州标旗光电科技发展股份有限公司 | Method for reducing measurement error of wide spectrum transmittance |
| CN112557362A (en) * | 2020-12-04 | 2021-03-26 | 厦门大学 | Synchronous fluorescence spectrum detection method using LED light source as continuous wave excitation light source |
| CN112782131A (en) * | 2019-11-11 | 2021-05-11 | 成都辰显光电有限公司 | Spectrum detection system and spectrum detection method |
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| CN105842221A (en) * | 2016-05-31 | 2016-08-10 | 北京金索坤技术开发有限公司 | Atomic fluorescence spectrometer and background subtraction method thereof |
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| CN103424358A (en) * | 2013-07-10 | 2013-12-04 | 中国科学院安徽光学精密机械研究所 | Multi-parameter temperature-controlled absorption cell device capable of simulating vertical atmospheric environment |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112782131A (en) * | 2019-11-11 | 2021-05-11 | 成都辰显光电有限公司 | Spectrum detection system and spectrum detection method |
| CN111829971A (en) * | 2020-08-10 | 2020-10-27 | 广州标旗光电科技发展股份有限公司 | Method for reducing measurement error of wide spectrum transmittance |
| CN112557362A (en) * | 2020-12-04 | 2021-03-26 | 厦门大学 | Synchronous fluorescence spectrum detection method using LED light source as continuous wave excitation light source |
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