CN104864962B - Method using spectrometer to synchronically measure light intensity and light quality - Google Patents

Method using spectrometer to synchronically measure light intensity and light quality Download PDF

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CN104864962B
CN104864962B CN201510274722.0A CN201510274722A CN104864962B CN 104864962 B CN104864962 B CN 104864962B CN 201510274722 A CN201510274722 A CN 201510274722A CN 104864962 B CN104864962 B CN 104864962B
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CN104864962A (en
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樊大勇
付增娟
徐文婷
谢宗强
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Institute of Botany of CAS
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Abstract

The invention relates to a method using a spectrometer to synchronically measure light intensity and light quality; the method comprises the following steps: selecting the spectrometer, and setting up a light energy value-light quantum flux density experience conversion model according to measure parameters of the selected spectrometer; using four LED light sources of different wavelength to combine with a standard light quantum meter so as to calibrate obtained spectrums, thus obtaining values of parameters k and C of the experience conversion module; using the selected spectrometer to synchronically measure and calculate so as to obtain the light intensity and light quality according to the obtained k and C value, and the light energy value-light quantum flux density experience conversion model, i.e., using the selected spectrometer to measure and calculate the light quantum flux density of any light source and light environment. The method is simple in operation, fast in calibration, reliable in measuring, suitable for promotion and application, thus having wide application prospect in the plant physiology, botany and ecology research fields.

Description

A kind of method of employing spectrogrph Simultaneous Determination light intensity and light quality
Technical field
The present invention relates to a kind of method for determining light intensity and light quality, adopts spectrogrph Simultaneous Determination light especially with regard to a kind of The method of strong and light quality.
Background technology
The impact of light intensity and light confrontation plant is constantly subjected to pay high attention in the research fields such as botany, ecology.From Varying level of the molecule to individuality, plant are responded and are adapted to the light intensity and light quality environment of change, and these processes are exactly green Color organ photosynthesis, individual growth, species regeneration coexist and syndynamic one of important inner principles.It is relevant The research of the impact of light intensity and light confrontation photosynthesis and plant growing is very more.By taking light intensity as an example, high light can cause Plant Light Close mechanism and photoinhibition occur, and then inhibit the carbon accumulation and individual growth of plant;The low light level can cause plant leaf blade to occur There is carbon hunger phenomenon and taper off in aetiolation, plant individual;By taking light quality as an example, the plant under complete stand in green glow and Far-red light is accounted under the luminous environment of main component, causes its photosynthetic utilization ratio to green glow higher than crown canopy plant, due to phytochrome The stem elongation growth of underground is caused to be higher than crown canopy plant the sensitivity of far-red light.In addition, light intensity and light confrontation are photosynthetic Effect has significant reciprocal action, and under such as light intensity identical blue and green light and HONGGUANG environment, the light of photosystem 2 (PS II) loses Sensitivity living highest under blue light, the Mn clusters that its reason may put oxygen protein complexes with II oxidant sides of PS are more biased towards absorbing blue light It is related.Obviously, in the related discipline research field such as plant physiology, ecological science, agricultural sciences, environmental science, to light intensity and It is an important job that light quality environment carries out accurate measurement.
Photon hypothesis (PPFD or PAR) refer to unit interval unit area as conventional light intensity quantizating index Upper incident light quantity subnumber, its unit are μm ol photons m-2s-1(or claim μm ol Einstein m-2s-1).400~ Light quantum in 700nm wave-length coverages can be electrochemical energy by chlorophyll molecule sorption enhanced, and for carbon assimilation mistake thereafter Journey.Although the light quantum energy of different wave length is different, for photosynthesis, the light quantum action effect of different wave length It is identical:After one blue light light quantum or HONGGUANG light quantum are by chlorophyll molecule absorption, the low-lying level electronics of chlorophyll molecule is swashed It is dealt into same excited state, the electronics of excited state subsequently carries out electron transmission and the conversion of photochemistry energy, and both unnecessary energy Heat energy is converted into then and can not be utilized again.Therefore it is most concerned with PAR in photosynthesis research, rather than unit area Light radiation flux (W m-2, or W m-2nm-1, energy unit).Other pigments such as carotenoid in chloroplast also 400~ 700nm wave-length coverages absorb light quantum, catch light with auxiliary and carry out the effect of heat dissipation process, this absorption also with light quantum Number rather than light radiation flux correlation.Additionally, research shows the far-red light of the blue violet light and more than 700nm of below 400nm to light With also having a major impact, such as Hakala et al. (2005) studies PSII light under the blue violet light for showing 200~400nm and loses for cooperation Sensitivity highest living, is the main cause of PSII Xanthophyll cycles.
At present, the measure of light intensity is using standardization light quantum meter (PAR pop one's head in, such as LI-190SB serial), but standardization Light quantum meter can only determine 400~700nm light intensity, it is impossible to determine blue violet light that spectral component can not be determined under 400nm and The far-red light of more than 700nm.The quantizating index of light quality generally adopts the unit area light radiation flux under specific wavelength, typically Use spectrophotometer light quality.Spectrogrph can determine spectral component, and its detectable wave-length coverage is very wide, such as certain optical fiber light The detectable wave-length coverage of spectrometer (AvaSpec-ULS2048 × 64) is 300~1100nm, is measured without the spectrogrph demarcated Data are typically the Counts under each wavelength (light energy value) or DN values, are the relative energy values for not having unit.Spectrogrph is not Light intensity i.e. photon hypothesis can be directly determined, and only reflects relative energy values without Counts the or DN values demarcated, its value Size and the time of integration, the quantum efficiency of detector, detector dark current etc. have substantial connection.Therefore, standardization light quantum meter Can not determine spectral component and its for the far-red light of blue violet light and more than 700nm under 400nm does not determine ability, and light Spectrometer but can not directly determine light intensity and be relative energy values without the spectrophotometer data demarcated, and the two can not same pacing Determine light intensity and light quality.
The content of the invention
For the problems referred to above, it is an object of the invention to provide a kind of using the synchronous accurate measure light source of spectrogrph and plant life The method of long light intensity and light quality environment.
For achieving the above object, the present invention takes technical scheme below:It is a kind of to adopt spectrogrph Simultaneous Determination light intensity and light The method of matter, it is characterised in that comprise the following steps:
1) chosen spectrum instrument, and the location parameter based on selected spectrogrph, set up light energy value close with light quantum flux The experience transformation model of degree:
In formula, PARcFor photon hypothesis, Counts is light energy value, QE(λ) the detector quantum for spectrogrph is imitated Rate, 1-λ of λ 2 are required wave-length coverage, and λ is wavelength;
2) light selected spectrogrph obtained using the LED light source combined standard light quantum meter of four different wave lengths Spectrum is demarcated, and obtains the value of parameter k and C in experience transformation model;
3) according to the transformation model for obtaining k and C values and light energy value and photon hypothesis, using the spectrum chosen Instrument Simultaneous Determination is calculated light quantity and light quality, i.e.,:By the spectrophotometer and calculating any light source and luminous environment of selection Photon hypothesis.
Further, also include calculating that using calculated k and C values the absolute light radiation corresponding to single Counts is led to Value, specially:The gross energy of the incident 1mol light quantum of per area per time is thenWherein, h is general Bright gram of constant, c are the light velocity, and λ is wavelength, and NA is Avogadro constant;For example:For PAR is 1000 μm of ol m-2s-1, wavelength be The HONGGUANG of 600nm, its gross energy are 199.6J m-2s-1, Counts value of the same light according under the conditions of is calculated according to parameter k and C Light radiation amount of flux for single Counts under 3842372, therefore 600nm light is 199.6/3842372 ≈ 5.2 × 10-5W m-2
Further, the step 2) using the LED light source combined standard light quantum meter of four different wave lengths to selected The spectrum obtained by spectrogrph is demarcated, and obtains the value of parameter k and C in experience transformation model, and detailed process is:
2.1) selection standard light quantum meter;
2.2) choose the LED light source of four different wave lengths;
2.3) light intensity of four LED light sources is adjusted respectively, makes each LED light source produce a series of light quantum flux close Degree, while each LED light source produces each photon hypothesis value, measures the LED using standard light quantum measurement Photon hypothesis PAR of the light source at the photon hypothesis valueλ, standard light quantum meter is removed after being measured, Same measurement position adopts spectral component Counts of the spectrophotometer LED light source at the photon hypothesis value;
2.4) obtain under single wavelength λ difference photo flux densities
2.5) pass throughWithBetween linear relationship chart be calculated parameter value k and C.
Further, the step 2.2) the middle narrow-band LED light source for choosing four different wave lengths, respectively including blue-ray LED light Source, green light LED light source, yellow light LED light source and red-light LED light source, wherein, the wavelength of blue light LED light source is 420~470nm, green The wavelength of light LED light source is 490~540nm, and the wavelength of yellow light LED light source is 550~600nm, and the wavelength of red-light LED light source is 640~690nm, the half-peak breadth of four kinds of waveform LED light sources are less than 24nm;Four kinds of waveform LED light source power are 10w, blue-ray LED The largest light intensity of light source is 400 μm of ol m-2s-1, the largest light intensity of green light LED light source is 800 μm of ol m-2s-1, yellow light LED light source Largest light intensity be 500 μm of ol m-2s-1, the largest light intensity of red-light LED light source is 1000 μm of ol m-2s-1
Due to taking above technical scheme, which has advantages below to the present invention:1st, the present invention is based on spectrophotometer parameter The experience transformation model of light energy value and photon hypothesis is set up, and the LED light source using four different wave lengths combines mark The spectrum obtained to spectrogrph by quasi-optical quantum meter is demarcated, and can calculate any wave-length coverage using calibrated spectrogrph Interior photon hypothesis, only need to be capable of achieving to experimental light sources and plant life using spectrogrph when experiment interior or field usage Long light intensity and light quality environment synchronize accurate measure, greatly widen the scope of application of spectrogrph.2nd, the present invention is using meter K the and C values for obtaining can calculate the absolute light/radiant flux value drawn corresponding to single Counts, therefore need not be using mark Quasi-optical source can obtain the absolute light/radiant flux value of unit Counts.The present invention is simple to operate, demarcate quick, determines reliable, For the research fields such as plant physiology, botany, ecology are with a wide range of applications.
Description of the drawings
Fig. 1 is that the quantum efficiency of the spectral component and spectrometer detector of four kinds of different wave length LED light sources of the present invention is bent Line, wherein, scheme the spectral component schematic diagram that (a) is four kinds of different wave length LED light sources, abscissa is wavelength, and unit is nm, indulges and sits Counts is designated as, the quantum efficiency curve synoptic diagram of (b) for spectrometer detector is schemed, abscissa is wavelength, and unit is nm, indulges and sits It is designated as quantum efficiency;
Fig. 2 is the PAR of the present inventionλWith CountsλRelation schematic diagram, abscissa are photon hypothesis, and unit is μm ol m-2s-1, vertical coordinate is accumulation Counts, wherein, scheme the accumulation Counts that (a) is that wavelength is 464nm blue lights, figure (b) is wavelength For the accumulation Counts of the green glow of 524nm, scheme (c) for wavelength for the gold-tinted of 573nm accumulation Counts;Scheming (d) for wavelength is The accumulation Counts of the HONGGUANG of 677nm;
Fig. 3 is the different wave length that the present invention is obtainedWithBetween linear relationship schematic diagram;
Fig. 4 is the spectrum of natural light in the embodiment of the present invention and determines PARcWith calculating PARmBetween relation schematic diagram, its In, it is the spectrogram using spectrophotometer natural light to scheme (a);Figure (b) is the PAR that calculated according to k and C values and use standard Graph of a relation between the PAR that light quantum meter is determined.
Specific embodiment
Detailed description is carried out to the present invention below in conjunction with accompanying drawing.It should be appreciated, however, that accompanying drawing has been provided only more Understand the present invention well, they should not be interpreted as limitation of the present invention.
Employing spectrogrph Simultaneous Determination light intensity and the method for light quality that the present invention is provided, comprise the following steps:
1st, chosen spectrum instrument, and selected spectrophotometer parameter is based on, set up light energy value and photon hypothesis Experience transformation model;
In the embodiment of the present invention, selected spectrogrph is the portable fiber-optic spectrometer of Dutch Avantes companies production, The model of the portable fiber-optic spectrometer:AvaSpec-ULS2048 × 64, AvaSpec-ULS2048 × 64 fiber spectrometer Parameter setting be time of integration 4ms, average time is 2 times, and the detector quantum efficiency of the spectrogrph is QE(λ)。
Spectrogrph is output as Counts (light energy value), and which is meant that the relative light radiation received by unit area CCD Amount of flux (energy of unit area), for visible ray, the light quantum energy of different wave length is different, can adopt following letter Number is indicated:
In formula, E is luminous energy, and unit is J;H is Planck's constant, and its value is 6.63 × 10-34J s;C is the light velocity, and its value is 3.0×108ms-1;λ is wavelength, and unit is m (in practical study, the unit of λ is nm).
The light radiation flux Counts of a certain specific wavelength λλThe light quantum of (light energy value) and per area per time Number PARλFunctional relation between (photon hypothesis):
As detector C CD of spectrogrph has the dark current of energy threshold, i.e. CCD, it is the detector constant of spectrogrph It is unrelated with wavelength, it is set as C (C is relevant with the dark current of the detector of spectrogrph), therefore, formula (2) can be changed into:
As spectrogrph has integrating effect, that is, absorbed luminous energy is tired out within a certain specific time of integration Product.Additionally, CCD itself has specific quantum efficiency QE, therefore the hc products that directly can not be given with theory, in this case Can be using constant k divided by quantum efficiency QE(λ) characterized.
Therefore, the PAR under certain wavelength XλWith CountsλBetween functional relationship be:
Can be seen that under certain wavelength X by formula (4), the Counts determined by spectrogrphλSurveyed with light quantum probe Fixed PARλBusiness be the constant for depending on λ.If different λ light can be determinedIn known QE(λ) then may be used under the conditions of To obtain the numerical value of k and C.After obtaining the numerical value of k and C, you can there is the light of the light of complex spectrum composition using spectrophotometer Quantum flux density, that is, to Counts in the range of 400~700nm of wavelengthλIt is integrated, based on spectrophotometer parameter Obtain the experience transformation model between light energy value and photon hypothesis:
In formula, PARcIt is photon hypothesis.It is of course also possible to profit is counted by changing the bound that formula (5) is integrated Calculate the PAR values of the light (such as the far-red light of blue violet light and more than 700nm under 400nm) in any wave-length coverage.
2nd, the light selected spectrogrph obtained using the LED light source combined standard light quantum meter of four different wave lengths Spectrum is demarcated, and obtains the value of parameter k and C in experience transformation model, and detailed process is:
1) selection standard light quantum meter
In the embodiment of the present invention, selected standard light quantum is calculated as the standard light quantum meter of Licor companies of U.S. production (LI190SB), for determining incident illumination photon hypothesis.
2) choose the narrow-band LED light source of four different wave lengths
Instrument producer is generally demarcated to spectrogrph using standard light source (usually Halogen light or deuterium lamp), but this The program of calibration process is complicated, and standard light source and determination of the environment is required strict.The blue light very narrow using half-peak breadth of the invention, Green glow, gold-tinted and red-light LED light source are used as Calibrating source.The different ripples of 4 of selected LED light source in the embodiment of the present invention It is long, respectively:, in the range of 420~470nm, the wavelength of green light LED light source is in 490~540nm models for the wavelength of blue light LED light source In enclosing, the wavelength of yellow light LED light source in the range of 550~600nm, the wavelength of red-light LED light source in the range of 640~690nm, The half-peak breadth of four kinds of waveform LED light sources is less than 24nm;Four kinds of waveform LED light source power are 10w, and blue light LED light source is most Big light intensity is 400 μm of ol m-2s-1, the largest light intensity of green light LED light source is 800 μm of ol m-2s-1, the maximum light of yellow light LED light source It is by force 500 μm of ol m-2s-1, the largest light intensity of red-light LED light source is 1000 μm of ol m-2s-1;Electricity needed for four kinds of waveform LED light sources Source should be 5v, and maximum current is 2A.As shown in Fig. 1 (a), the peak value of the LED light source corresponding to blue and green light, gold-tinted and HONGGUANG Wavelength is 464,524,573 and 677nm, and half-peak breadth is respectively 20,24,14 and 6nm.Due to selected four LED light sources half Peak width is very narrow, is especially suitable for carrying out calibration experiment.As shown in Fig. 1 (b), exist in 200~800nm wave-length coverage internal quantum efficiencies Fluctuate in the range of 0.5 to 0.8, drastically decline more than the quantum efficiency of the detector of 800nm spectrogrphs.
3) light intensity of four LED light sources is adjusted respectively, makes each LED light source produce a series of light quantum flux close Degree, produces at each photon hypothesis value in each LED light source, measures the LED light source using standard light quantum measurement Photon hypothesis PAR at the photon hypothesis valueλ, standard light quantum meter is removed after being measured, same Measurement position adopts spectral component Counts of the spectrophotometer LED light source at the photon hypothesis value.
4) obtain under single wavelength λ difference photo flux density gradients
As shown in Fig. 2 a series of photon hypothesis are set under single wavelength LED light source, it is possible to obtain PARλWith CountsλBetween relation.Due to PARλIt is photon hypothesis of the standard light quantum meter to 400~700nm of wave-length coverage Measurement result, therefore (AC is integrated to the Counts in the wave-length coverageλ).Due to this four LED half-peak breadth very It is narrow, it is believed that ACλIt is equal to Countsλ
From figure 2 it can be seen that under single wavelength LED light source CountsλWith PARλWith good proportionate relationship, such as Shown in Fig. 2 (a)~(d), CountsλWith PARλBusiness be respectively 7648 (464nm blue lights), 5690 (524nm green glows), 4275 (yellow Light, 573nm) and 2841 (HONGGUANG, 677nm), the correctness of formula (4) is fully demonstrated, while also illustrating what spectrogrph was measured Numerical value Counts its essence is relative light radiation amount of flux, if Counts is photon hypothesis value, Countsλ With PARλBusiness will not change with the change of wavelength.
5) pass throughWithBetween linear relationship chart be calculated parameter value k and C.
It is the different wave length obtained according to formula (4) as shown in Figure 3WithBetween linear relationship chart, Can calculate that by Fig. 3 the k value for obtaining the spectrogrph is that 5322042.5, C values are -7677.2 (PARλWith a μm ol m-2s-1For unit, λ In units of nm).
As extraneous light intensity affects larger to the scaling method, thus the present invention should indoors under low light condition (<1μ mol m-2m-1) demarcated, the nominal time can be completed in 30 minutes, but not limited to this.
3rd, according to the transformation model for obtaining k and C values and light energy value and photon hypothesis, using the spectrum chosen Instrument Simultaneous Determination is calculated light quantity and light quality, i.e.,:According to adopt by choose spectrophotometer and calculate any light source and The photon hypothesis of luminous environment.
In a preferred embodiment, the present invention can also adopt k and C values to calculate absolute corresponding to single Counts Light radiation amount of flux (absolute value that light energy can be worth), specially:Can be by formula for the energy of a photon of specific wavelength (1) obtain, therefore the gross energy of the incident 1mol light quantum of per area per time is thenWherein, NA be Ah Fu Jiade constants, its numerical value are 6.02 × 1023.For PAR is 1000 μm of ol m-2s-1, wavelength for 600nm HONGGUANG, its total energy Measure as 199.6J m-2s-1(=W m-2).It is 3842372 that Counts value of the same light according under the conditions of is calculated according to parameter k and C, Therefore under 600nm light, the light radiation amount of flux of single Counts should be 199.6/3842372 ≈ 5.2 × 10-5W m-2, according to This, without the need for the relative light radiation flux unit of spectrum being converted into absolute light/radiant flux unit with standard light source.
The employing spectrogrph Simultaneous Determination light intensity present invention provided below by specific embodiment and the method for light quality Correctness is verified that concrete proof procedure is:
Tested using the natural light with complex spectrum composition, natural light is determined as 11 points or so open airs of fine day noon Mensuration mode, incident intensity of being decayed by way of outer housing multilamellar gauze produce a series of photon hypothesis.Using AvaSpec-ULS2048 × 64 fiber spectrometer determines the spectral component of natural light, and calculates PAR according to k and C;Adopt simultaneously Standard light quantum meter determines actual PAR, calculates the accuracy of PAR by comparison test.If Fig. 4 (a) is to adopt fiber spectrometer Determine the spectrogram of natural light, Fig. 4 (b) be the PAR and PAR with standard light quantum meter measure that calculated according to k and C values it Between graph of a relation, by Fig. 4 (b) as can be seen that calculating that the PAR that the PAR that obtains is obtained with measurement has fine linear relationship, its phase Relation number is close to 1, and calculates that PAR, in -2% to+5% range of error of measured value, has absolutely proved k and C parameter values Reliability, effectively demonstrates the present invention by directly determining the correct of photon hypothesis and light quality using spectrogrph is synchronous Property.
The various embodiments described above are merely to illustrate the present invention, and wherein each implementation steps of each method etc. all can be to be varied from , every equivalents carried out on the basis of technical solution of the present invention and improvement should not exclude the protection in the present invention Outside scope.

Claims (2)

1. a kind of method of employing spectrogrph Simultaneous Determination light intensity and light quality, it is characterised in that comprise the following steps:
1) chosen spectrum instrument, and the location parameter based on selected spectrogrph, set up light energy value with photon hypothesis Experience transformation model:
PAR &lambda; = &Integral; &lambda; 1 &lambda; 2 Counts &lambda; &lambda;Q E ( &lambda; ) k + C&lambda;Q E ( &lambda; ) d &lambda;
In formula, PARλFor photon hypothesis, CountsλFor the light energy value of a certain specific wavelength λ, QE(λ) it is spectrogrph Detector quantum efficiency, 1-λ of λ 2 are required wave-length coverage, and λ is wavelength;
2) spectrum obtained by selected spectrogrph is entered using the LED light source combined standard light quantum meter of four different wave lengths Rower is fixed, obtains the value of parameter k and C in experience transformation model, and detailed process is:
2.1) selection standard light quantum meter;
2.2) choose the LED light source of four different wave lengths;
2.3) light intensity of four LED light sources is adjusted respectively, makes each LED light source produce a series of photon hypothesis, While each LED light source produces each photon hypothesis value, the LED light source is measured using standard light quantum measurement Photon hypothesis PAR at the photon hypothesis valueλ, standard light quantum meter is removed after being measured, same Measurement position adopts light energy value Counts of the spectrophotometer LED light source at the photon hypothesis valueλ
2.4) obtain under single wavelength λ difference photo flux densities
2.5) pass throughWithBetween linear relationship chart be calculated parameter value k and C;
3) it is according to the transformation model for obtaining k and C values and light energy value and photon hypothesis, same using the spectrogrph chosen Step measure and calculation obtains light quantity and light quality, i.e.,:By the spectrophotometer chosen and the light quantity for calculating any light source and luminous environment Sub- flux density.
2. the method for a kind of employing spectrogrph Simultaneous Determination light intensity as claimed in claim 1 and light quality, it is characterised in that:It is described Step 2.2) choose four different wave lengths narrow-band LED light source, respectively including blue light LED light source, green light LED light source, yellow light LED Light source and red-light LED light source, wherein, the wavelength of blue light LED light source is 420~470nm, the wavelength of green light LED light source is 490~ 540nm, the wavelength of yellow light LED light source is 550~600nm, and the wavelength of red-light LED light source is 640~690nm, four kinds of wavelength LED The half-peak breadth of light source is less than 24nm;Four kinds of waveform LED light source power are 10w, and the largest light intensity of blue light LED light source is 400 μ mol m-2s-1, the largest light intensity of green light LED light source is 800 μm of ol m-2s-1, the largest light intensity of yellow light LED light source is 500 μm of ol m-2s-1, the largest light intensity of red-light LED light source is 1000 μm of ol m-2s-1
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