CN104864962A - Method using spectrometer to synchronically measure light intensity and light quality - Google Patents
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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
Technical field
The present invention relates to a kind of method measuring light intensity and light quality, particularly about a kind of method adopting spectrometer Simultaneous Determination light intensity and light quality.
Background technology
Light intensity and light quality is shown great attention in the research field such as botany, ecology the impact of plant always.Varying level from molecule to individuality, the light intensity and light quality environment of plant to change responds and adapts to, and these processes just green organs photosynthesis, individual growth, species regeneration coexist and one of the important inner principles of syndynamic.About the research of light intensity and light quality on the impact of photosynthesis and plant growth is very many.For light intensity, high light can cause Photosynthetic Apparatus in Plant to occur photoinhibition, and then inhibits carbon accumulation and the individual growth of plant; The low light level can cause plant leaf blade to occur aetiolation, and plant individual occurs carbon hunger phenomenon and tapers off; For light quality, plant under complete stand is under green glow and far-red light account for the luminous environment of principal ingredient, cause it to the photosynthetic utilization ratio of green glow higher than crown canopy plant, because the susceptibility of phytochrome to far-red light makes the stem elongation growth of underground be higher than crown canopy plant.In addition, light intensity and light quality has significant reciprocation to photosynthesis, under the blue light that such as light intensity is identical, green glow and ruddiness environment, the light inactivation susceptibility of photosystem 2 (PS II) is the highest under blue light, and its reason may be oxidized Mn bunch of being sidelong oxygen protein complexes to PS II and more be partial to that to absorb blue light relevant.Obviously, in related discipline research fields such as plant physiology, ecological science, agricultural sciences, environmental sciences, carrying out accurately measuring to light intensity and light quality environment is an important job.
Photon hypothesis (PPFD or PAR) is as conventional light intensity quantizating index, and refer to light quantity subnumber incident in unit interval unit area, its unit is a μm ol photons m
-2s
-1(or claim μm ol Einstein m
-2s
-1).Light quantum in 400 ~ 700nm wavelength coverage can be electrochemical energy by chlorophyll molecule sorption enhanced, and for carbon assimilation process thereafter.Although the light quantum energy of different wave length is different, but for photosynthesis, the light quantum action effect of different wave length is identical: after a blue light light quantum or ruddiness light quantum are absorbed by chlorophyll molecule, by the low-lying level electron excitation of chlorophyll molecule to same excited state, the electronics of excited state carries out electron transmission and photochemistry Conversion of Energy subsequently, and both are then converted into heat energy and can not be utilized again by unnecessary energy.Therefore in photosynthesis research it is of concern that PAR, instead of light/radiation flux (W m of unit area
-2, or W m
-2nm
-1, energy unit).Other pigment such as the carotenoid in chloroplast also absorbs light quantum in 400 ~ 700nm wavelength coverage, and have the auxiliary effect of catching light and carrying out heat dissipation process, this absorption is also relevant to light quantity subnumber instead of light/radiation flux.In addition, research shows that the royal purple light of below 400nm and the far-red light of more than 700nm also have material impact to photosynthesis, as Hakalaet al. (2005) research show the royal purple light of 200 ~ 400nm under PSII light inactivation susceptibility the highest, be the main cause of PSII Xanthophyll cycle.
At present, the mensuration of light intensity adopts standardization light quantum meter (PAR probe, as LI-190SB series), but standardization light quantum meter only can measure 400 ~ 700nm light intensity, can not measure the far-red light that spectral component can not measure royal purple light under 400nm and more than 700nm.The quantizating index of light quality adopts the unit area light/radiation flux under specific wavelength usually, generally uses spectrophotometer light quality.Spectrometer can measure spectral component, its detectable wavelength coverage is very wide, such as certain fiber spectrometer (AvaSpec-ULS2048 × 64) detectable wavelength coverage is 300 ~ 1100nm, Counts (light energy value) under the normally each wavelength of the data that not proven spectrometer records or DN value are the relative energy values not having unit.Spectrometer can not directly measure light intensity and photon hypothesis, and not proven Counts or DN value only reflects relative energy values, and the quantum efficiency, detector dark current etc. of its value size and integral time, detector have substantial connection.Therefore, standardization light quantum meter can not measure spectral component and its far-red light for the royal purple light under 400nm and more than 700nm does not measure ability, and spectrometer directly can not measure light intensity and not proven spectrophotometer data are relative energy values, the two all can not Simultaneous Determination light intensity and light quality.
Summary of the invention
For the problems referred to above, the object of this invention is to provide a kind of method adopting the light intensity and light quality environment of the synchronous Accurate Measurement light source of spectrometer and plant growth.
For achieving the above object, the present invention takes following technical scheme: a kind of method adopting spectrometer Simultaneous Determination light intensity and light quality, is characterized in that comprising the following steps:
1) chosen spectrum instrument, and based on the location parameter of selected spectrometer, set up the experience transformation model of light energy value and photon hypothesis:
In formula, PAR
cfor photon hypothesis, Counts is light energy value, Q
e(λ) be the detecting device quantum efficiency of spectrometer, λ 1-λ 2 is required wavelength coverage, and λ is wavelength;
2) adopt the LED light source combined standard light quantum meter of four different wave lengths to demarcate the spectrum that selected spectrometer obtains, obtain the value of parameter k and C in experience transformation model;
3) according to the transformation model obtaining k and C value and light energy value and photon hypothesis, the spectrometer Simultaneous Determination chosen is adopted to calculate light quantity and light quality, that is: by the spectrophotometer chosen and the photon hypothesis calculating any light source and luminous environment.
Further, also comprise the absolute light/radiant flux value of k and C value reckoning corresponding to single Counts adopting and calculate, be specially: the gross energy of the 1mol light quantum of per area per time incidence is then
wherein, h is Planck's constant, and c is the light velocity, and λ is wavelength, and NA is Avogadro constant; Such as: be 1000 μm of ol m for PAR
-2s
-1, wavelength is the ruddiness of 600nm, its gross energy is 199.6J m
-2s
-1, be 3842372 according to the Counts value that parameter k and C calculates under identical illumination condition, therefore under 600nm light, the light/radiant flux value of single Counts is 199.6/3842372 ≈ 5.2 × 10
-5w m
-2.
Further, described step 2) adopt the LED light source combined standard light quantum meter of four different wave lengths to demarcate the spectrum that selected spectrometer obtains, obtain the value of parameter k and C in experience transformation model, detailed process is:
2.1) selection standard light quantum meter;
2.2) LED light source of four different wave lengths is chosen;
2.3) light intensity of four LED light sources is regulated respectively, each LED light source is made all to produce a series of photon hypothesis, while each LED light source produces each photon hypothesis value, the measurement of standard light quantum is adopted to measure the photon hypothesis PAR of this LED light source at this photon hypothesis value place
λ, remove standard light quantum meter after having measured, adopt the spectral component Counts of this LED light source of spectrophotometer at this photon hypothesis value place in same measuring position;
2.4) under the different photo flux density of acquisition single wavelength λ
2.5) pass through
with
between linear relationship chart calculate parameter value k and C.
Further, described step 2.2) in choose the narrow-band LED light source of four different wave lengths, comprise blue light LED light source, green light LED light source, yellow light LED light source and red-light LED light source respectively, wherein, the wavelength of blue light LED light source is 420 ~ 470nm, and the wavelength of green light LED light source is 490 ~ 540nm, and the wavelength of yellow light LED light source is 550 ~ 600nm, the wavelength of red-light LED light source is 640 ~ 690nm, and the half-peak breadth of four kinds of waveform LED 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 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 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.
The present invention is owing to taking above technical scheme, it has the following advantages: 1, the present invention is based on the experience transformation model that spectrophotometer parameter sets up light energy value and photon hypothesis, and adopt the LED light source combined standard light quantum meter of four different wave lengths to demarcate the spectrum that spectrometer obtains, adopt calibrated spectrometer can calculate photon hypothesis in any wavelength coverage, in laboratory or field only need adopt spectrometer can realize carrying out synchronous Accurate Measurement to the light intensity and light quality environment of experimental light sources and plant growth when using, greatly widen the scope of application of spectrometer.2, the present invention adopts k and the C value calculated can calculate the absolute light/radiant flux value drawn corresponding to single Counts, the absolute light/radiant flux value therefore without the need to adopting standard sources can obtain unit Counts.The present invention is simple to operate, demarcation is quick, measures reliable, is with a wide range of applications for research fields such as plant physiology, botany, ecology.
Accompanying drawing explanation
Fig. 1 is the spectral component of four kinds of different wave length LED light sources of the present invention and the quantum efficiency curve of spectrometer detector, wherein, figure (a) is the spectral component schematic diagram of four kinds of different wave length LED light sources, horizontal ordinate is wavelength, and unit is nm, and ordinate is Counts, the quantum efficiency curve synoptic diagram that figure (b) is spectrometer detector, horizontal ordinate is wavelength, and unit is nm, and ordinate is quantum efficiency;
Fig. 2 is PAR of the present invention
λwith Counts
λrelation schematic diagram, horizontal ordinate is photon hypothesis, and unit is a μm ol m
-2s
-1ordinate is accumulation Counts, wherein, figure (a) for wavelength be the accumulation Counts of 464nm blue light, figure (b) for wavelength be the accumulation Counts of the green glow of 524nm, figure (c) for wavelength be the accumulation Counts of the gold-tinted of 573nm; Figure (d) for wavelength be the accumulation Counts of the ruddiness of 677nm;
Fig. 3 is the different wave length that the present invention obtains
with
between linear relationship schematic diagram;
Fig. 4 is the spectrum of natural light in the embodiment of the present invention and measures PAR
cwith calculating PAR
mbetween relation schematic diagram, wherein, figure (a) is the spectrogram adopting spectrophotometer natural light; Figure (b) is the PAR that calculates according to k and C value and measures the graph of a relation between fixed PAR with standard light quantum.
Embodiment
Below in conjunction with accompanying drawing, detailed description is carried out to the present invention.But should be appreciated that being provided only of accompanying drawing understands the present invention better, they not should be understood to limitation of the present invention.
The method of employing spectrometer Simultaneous Determination light intensity and light quality provided by the invention, comprises the following steps:
1, chosen spectrum instrument, and based on selected spectrophotometer parameter, set up the experience transformation model of light energy value and photon hypothesis;
Spectrometer selected in the embodiment of the present invention is the portable fiber-optic spectrometer that Dutch Avantes company produces, the model of this portable fiber-optic spectrometer is: AvaSpec-ULS2048 × 64, the setting parameter of AvaSpec-ULS2048 × 64 fiber spectrometer is 4ms integral time, average time is 2 times, and the detecting device quantum efficiency of this spectrometer is Q
e(λ).
The output of spectrometer is Counts (light energy value), its implication is light/radiant flux value relatively (energy of unit area) that unit area CCD receives, for visible ray, the light quantum energy of different wave length is different, can adopt and represent with minor function:
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 × 10
8ms
-1; λ is wavelength, and unit is m (in practical study, the unit of λ is nm).
Light/radiation flux the Counts of a certain specific wavelength λ
λthe light quantity subnumber PAR of (light energy value) and per area per time
λfunctional relation between (photon hypothesis):
Because the detector C CD of spectrometer has energy threshold, i.e. the dark current of CCD, be that detecting device constant and the wavelength of spectrometer has nothing to do, be set as C (C is relevant with the dark current of the detecting device of spectrometer), therefore, formula (2) can become:
Because spectrometer has integrating effect, namely within a certain specific integral time, absorbed luminous energy is accumulated.In addition, CCD itself has specific quantum efficiency Q
e, therefore can not, directly with theoretical given hc product, constant k can be adopted in this case divided by quantum efficiency Q
e(λ) characterize.
Therefore, the PAR under certain wavelength X
λwith Counts
λbetween funtcional relationship be:
As can be seen from formula (4), under certain wavelength X, the Counts that spectrometer measures
λto pop one's head in the PAR measured with light quantum
λbusiness be the constant depending on λ.If different λ light can be measured
at known Q
e(λ) numerical value of k and C can then be obtained under condition.Obtain k and C numerical value after, spectrophotometer can be adopted to have the photon hypothesis of the light of complex spectrum composition, namely within the scope of wavelength 400 ~ 700nm to Counts
λcarry out integration, obtain the experience transformation model between light energy value and photon hypothesis based on spectrophotometer parameter:
In formula, PAR
cit is photon hypothesis.Certainly, the PAR value of light in any wavelength coverage (far-red light as the royal purple light under 400nm and more than 700nm) also can be calculated in order to the bound of amendment formula (5) integration.
2, adopt the LED light source combined standard light quantum meter of four different wave lengths to demarcate the spectrum that selected spectrometer obtains, obtain the value of parameter k and C in experience transformation model, detailed process is:
1) selection standard light quantum meter
Standard light quantum selected in the embodiment of the present invention counts the standard light quantum meter (LI190SB) that Licor company of the U.S. produces, for measuring incident light photon hypothesis.
2) the narrow-band LED light source of four different wave lengths is chosen
Instrument producer adopts standard sources (being generally Halogen lamp LED or deuterium lamp) to demarcate spectrometer usually, but the program of this calibration process is complicated, and to standard sources and measure environmental requirement strict.The blue light that the present invention adopts half-peak breadth very narrow, green glow, gold-tinted and red-light LED light source are as Calibrating source.4 different wave lengths of LED light source selected in the embodiment of the present invention, be respectively: the wavelength of blue light LED light source is within the scope of 420 ~ 470nm, the wavelength of green light LED light source is within the scope of 490 ~ 540nm, the wavelength of yellow light LED light source is within the scope of 550 ~ 600nm, the wavelength of red-light LED light source is within the scope of 640 ~ 690nm, and the half-peak breadth of four kinds of waveform LED 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 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 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; Needed for four kinds of waveform LED light source, power supply should be 5v, and maximum current is 2A.As shown in Fig. 1 (a), the peak wavelength of the LED light source corresponding to blue light, green glow, gold-tinted and ruddiness is 464,524,573 and 677nm, and half-peak breadth is respectively 20,24,14 and 6nm.Because selected four LED light source half-peak breadths are all very narrow, be applicable to very much carrying out calibration experiment.As shown in Fig. 1 (b), fluctuate in 0.5 to 0.8 scope at 200 ~ 800nm wavelength coverage internal quantum efficiency, sharply decline more than the quantum efficiency of the detecting device of 800nm spectrometer.
3) light intensity of four LED light sources is regulated respectively, each LED light source is made all to produce a series of photon hypothesis, produce each photon hypothesis value place at each LED light source, adopt the measurement of standard light quantum to measure the photon hypothesis PAR of this LED light source at this photon hypothesis value place
λ, remove standard light quantum meter after having measured, adopt the spectral component Counts of this LED light source of spectrophotometer at this photon hypothesis value place in same measuring position.
4) under the different photo flux density gradient of acquisition single wavelength λ
As shown in Figure 2, under single wavelength LED light source, set a series of photon hypothesis, can PAR be obtained
λwith Counts
λbetween relation.Due to PAR
λbe standard light quantum meter to the measurement result of the photon hypothesis of wavelength coverage 400 ~ 700nm, therefore integration (AC has been carried out to the Counts in this wavelength coverage
λ).Because the half-peak breadth of these four LED is all very narrow, AC can be thought
λbe equal to Counts
λ.
As can be seen from Figure 2, Counts under single wavelength LED light source
λwith PAR
λthere is good proportionate relationship, as shown in Fig. 2 (a) ~ (d), Counts
λwith PAR
λbusiness be respectively 7648 (464nm blue lights), 5690 (524nm green glows), 4275 (gold-tinteds, 573nm) He 2841 (ruddiness, 677nm), fully demonstrate the correctness of formula (4), also illustrate that its essence of numerical value Counts that spectrometer records is light/radiant flux value relatively simultaneously, if Counts is photon hypothesis value, Counts
λwith PAR
λbusiness can not change with the change of wavelength.
5) pass through
with
between linear relationship chart calculate parameter value k and C.
Be illustrated in figure 3 the different wave length that obtains according to formula (4)
with
between linear relationship chart, can calculate that the k value obtaining this spectrometer be 5322042.5, C value is-7677.2 (PAR by Fig. 3
λwith a μm olm
-2s
-1for unit, λ is in units of nm).
Because extraneous light intensity is comparatively large on the impact of this scaling method, therefore the present invention should under indoor weak light condition (<1 μm of olm
-2m
-1) demarcate, the nominal time can complete in 30 minutes, but was not limited thereto.
3, according to the transformation model obtaining k and C value and light energy value and photon hypothesis, the spectrometer Simultaneous Determination chosen is adopted to calculate light quantity and light quality, that is: according to the spectrophotometer adopted by choosing and the photon hypothesis calculating any light source and luminous environment.
In a preferred embodiment, the present invention can also adopt k and C value to calculate absolute light/radiant flux value corresponding to the single Counts absolute value of value (luminous energy can), be specially: the energy for a photon of specific wavelength can obtain by formula (1), and the gross energy of the 1mol light quantum of therefore per area per time incidence is then
wherein, NA is Avogadro constant, and its numerical value is 6.02 × 10
23.Be 1000 μm of ol m for PAR
-2s
-1, wavelength is the ruddiness of 600nm, its gross energy is 199.6J m
-2s
-1(=W m
-2).The Counts value calculated under identical illumination condition according to parameter k and C is 3842372, and therefore under 600nm light, the light/radiant flux value of single Counts should be 199.6/3842372 ≈ 5.2 × 10
-5w m
-2, accordingly, without the need to can be absolute light/radiation flux unit by the light relatively/radiation flux unit conversion of spectrum by standard sources.
Verify below by the correctness of specific embodiment to the method for employing spectrometer Simultaneous Determination light intensity and light quality provided by the invention, concrete proof procedure is:
Adopt the natural light with complex spectrum composition to test, natural light is determined as fine day about 11 outdoor mensuration modes at noon, by the mode attenuate incident light intensity of outer cover multilayer gauze, produces a series of photon hypothesis.Adopt AvaSpec-ULS2048 × 64 fiber spectrometer to measure the spectral component of natural light, and calculate PAR according to k and C; Adopt the fixed actual PAR of standard light quantum measurement simultaneously, calculated the accuracy of PAR by comparison test.If Fig. 4 (a) is the spectrogram adopting fiber spectrometer to measure natural light, Fig. 4 (b) is the PAR that calculates according to k and C value and measures the graph of a relation between fixed PAR with standard light quantum, can be found out by Fig. 4 (b), the PAR that the PAR that reckoning obtains obtains with measurement has fine linear relationship, its related coefficient is close to 1, and calculate that PAR is in-2% to+5% error range of measured value, absolutely prove the reliability of k and C parameter value, effectively demonstrate the correctness of the present invention by adopting spectrometer synchronously directly to measure photon hypothesis and light quality.
The various embodiments described above are only for illustration of the present invention, and wherein each implementation step etc. of each method all can change to some extent, and every equivalents of carrying out on the basis of technical solution of the present invention and improvement, all should not get rid of outside protection scope of the present invention.
Claims (4)
1. adopt a method for spectrometer Simultaneous Determination light intensity and light quality, it is characterized in that comprising the following steps:
1) chosen spectrum instrument, and based on the location parameter of selected spectrometer, set up the experience transformation model of light energy value and photon hypothesis:
In formula, PAR
cfor photon hypothesis, Counts is light energy value, Q
e(λ) be the detecting device quantum efficiency of spectrometer, λ 1-λ 2 is required wavelength coverage, and λ is wavelength;
2) adopt the LED light source combined standard light quantum meter of four different wave lengths to demarcate the spectrum that selected spectrometer obtains, obtain the value of parameter k and C in experience transformation model;
3) according to the transformation model obtaining k and C value and light energy value and photon hypothesis, the spectrometer Simultaneous Determination chosen is adopted to calculate light quantity and light quality, that is: by the spectrophotometer chosen and the photon hypothesis calculating any light source and luminous environment.
2. a kind of method adopting spectrometer Simultaneous Determination light intensity and light quality as claimed in claim 1, it is characterized in that: also comprise the absolute light/radiant flux value of k and C value reckoning corresponding to single Counts adopting and calculate, be specially: the gross energy of the 1mol light quantum of per area per time incidence is then
wherein, h is Planck's constant, and c is the light velocity, and λ is wavelength, and NA is Avogadro constant; Such as: be 1000 μm of ol m for PAR
-2s
-1, wavelength is the ruddiness of 600nm, its gross energy is 199.6J m
-2s
-1, be 3842372 according to the Counts value that parameter k and C calculates under identical illumination condition, therefore under 600nm light, the light/radiant flux value of single Counts is 199.6/3842372 ≈ 5.2 × 10
-5w m
-2.
3. a kind of method adopting spectrometer Simultaneous Determination light intensity and light quality as claimed in claim 1 or 2, it is characterized in that: described step 2) adopt the LED light source combined standard light quantum meter of four different wave lengths to demarcate the spectrum that selected spectrometer obtains, the value of parameter k and C in acquisition experience transformation model, detailed process is:
2.1) selection standard light quantum meter;
2.2) LED light source of four different wave lengths is chosen;
2.3) light intensity of four LED light sources is regulated respectively, each LED light source is made all to produce a series of photon hypothesis, while each LED light source produces each photon hypothesis value, the measurement of standard light quantum is adopted to measure the photon hypothesis PAR of this LED light source at this photon hypothesis value place
λ, remove standard light quantum meter after having measured, adopt the spectral component Counts of this LED light source of spectrophotometer at this photon hypothesis value place in same measuring position;
2.4) under the different photo flux density of acquisition single wavelength λ
2.5) pass through
with
between linear relationship chart calculate parameter value k and C.
4. a kind of method adopting spectrometer Simultaneous Determination light intensity and light quality as claimed in claim 3, it is characterized in that: described step 2.2) choose the narrow-band LED light source of four different wave lengths, comprise blue light LED light source, green light LED light source, yellow light LED light source and red-light LED light source respectively, 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, the wavelength of red-light LED light source is 640 ~ 690nm, and the half-peak breadth of four kinds of waveform LED 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 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 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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