CN103868901B - Based on phytoplankton identification assay method and the device of discrete three-dimensional fluorescence spectrum - Google Patents
Based on phytoplankton identification assay method and the device of discrete three-dimensional fluorescence spectrum Download PDFInfo
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Abstract
The invention discloses a kind of phytoplankton identification assay method based on discrete three-dimensional fluorescence spectrum and device, assay method is according to the fingerprint characteristic of phytoplankton fluorescence spectrum, select different photosynthetic pigments characteristic of correspondence fluorescence peak as analytic target, farthest remain Phytoplankton Characteristic information in Three-Dimensional Fluorescence Spectra, with each characteristic spectrum in normal scatter three-dimensional fluorescence spectrum storehouse, matching is carried out to sample characteristic spectrum, be finally inversed by phytoplankton species and content thereof in sample; Determinator using multiband LED array as excitation source, using multiband filter set as receiving the band selector of fluorescence.Apparatus of the present invention are simple and compact for structure, volume is little, easy to carry, be suitable for the identification for on-the-spot phytoplankton and mensuration, in conjunction with invention in propose phytoplankton identification and assay method, phytoplankton scene category measurement fast can be realized.
Description
Technical field
The present invention relates to environmental science, be specifically related to a kind of phytoplankton identification assay method based on discrete three-dimensional fluorescence spectrum and device.
Background technology
Phytoplankton species and quantity can reflect the ecologic regime in this region, phytoplankton is just used as the indicator organism of water quality for a long time, special in water field of big area, measure species and the quantity of phytoplankton, have even more important Practical significance to water quality assessment.Along with China's inland lake and offshore sea waters body eutrophication, cause water phytoplankton excessive proliferation, red tide and wawter bloom take place frequently, and can carry out phytoplankton species and data analysis technique and equipment directly, fast, for red tide and wawter bloom monitoring and warning provide important means in the urgent need to development.
The identification assay method that phytoplankton is conventional mainly contains high performance liquid chromatography (HPLC), spectrophotometric method, fluorometry.Spectrophotometric method and HPLC have that selectivity is good, the feature such as separating property and sensitivity, in effluent monitoring analysis and the lake eutrophication investigation widespread use of China, wherein HPLC is U.S. EPA Plays planktonic algae chlorophyll-a concentration detection method 447.0, but this two prescribing method all needs the detecting instrument of complexity and costliness and sample needs pre-service, analysis time is long, equipment set volume is large, high to experimental situation conditional request, be only suitable for measuring in laboratory; Fluorometry have highly sensitive, be easy to realize real-time live and detect and material had to advantages such as good distinctive and extensively concerned, be that the most potential phytoplankton identifies mensuration means fast.Cowles etc. (1993), by measuring the displacement of phycoerythrin fluorescence emission spectrum wavelength, have inferred the change containing phycoerythrin phytoplankton species and abundance in seawater; The feature fluorescence excitation spectrum that Lee etc. (1995) send according to the phycocyanin that blue-green algae has, establishes the fluorescence analysis of on-the-spot monitored in vivo blue-green algae content; Kolbowski etc. (1995) have separated three main algal populations by initial chlorophyll phosphor region; Beutler etc. (2002), utilize phytoplankton Chlorophyll in vivo fluorescence excitation spectrum, are divided into by phytoplankton four large classes (green alga, blue-green algae, hidden algae, mixed algae (containing dinoflagellate and diatom)) to identify and measure, establish phytoplankton composition measuring technology.But, independent utilization excites or emitting fluorescence spectrum carries out phytoplankton category measurement, the defect existed is that identification is inadequate with estimating precision, pigment cannot be formed close phytoplankton (as Bacillariophyta and Pyrrophyta) or with the different types of phytoplankton Classification and Identification of class, be difficult to the needs meeting China's red tide and the common phytoplankton monitoring of wawter bloom.(2007), Su Rongguo etc. (2008) such as Zhang Qianqian etc. (2006), Wang Zhigang utilize phytoplankton live body three-dimensional fluorescence spectrum, in conjunction with the spectral model such as parallel factor, wavelet transformation algorithm, have carried out category measurement to the concentration of phytoplankton.Because three-dimensional fluorescence spectrum provides the finger print information of more horn of plenty, so can distinguish phytoplankton in more sub-layers time, phytoplankton identification and estimating precision have and largely improve.But because the mechanisms such as conventional three-dimensional fluorescence spectral measuring instrument volume is large, power consumption is high, grating transmission require high to operation stability, make the phytoplankton identification assay method based on three-dimensional fluorescence spectrum rest on the laboratory sample analysis phase, be difficult to the demand meeting phytoplankton on-site identification mensuration.
Goal of the invention
For China's body eutrophication present situation and country to the urgent demand of red tide and wawter bloom disaster monitoring, in conjunction with the developing direction of phytoplankton method and technique, the present invention proposes a kind of phytoplankton identification assay method based on discrete three-dimensional fluorescence spectrum, and for phytoplankton fluorescence spectral characteristic, devise a kind of discrete three-dimensional fluorescence spectrum determinator, this device volume is little, low in energy consumption, simple and stable structure, can the discrete three-dimensional fluorescence spectrum of feature of Site Detection phytoplankton, realize phytoplankton species identification and assay.
Summary of the invention
The invention provides a kind of phytoplankton identification assay method based on discrete three-dimensional fluorescence spectrum and device, directly can measure group's composition and the pigment content of phytoplankton in water body, whole operating process, without the need to pre-service, can be used for the on-the-spot fast investigation of field water phytoplankton.
The technical solution used in the present invention is:
Based on a phytoplankton identification assay method for discrete three-dimensional fluorescence spectrum, it is characterized in that, specifically comprise the following steps:
(1) the continuous three-dimensional fluorescence spectrum of the different class phytoplanktons of fluorescence spectrophotometer measurement is first utilized, the fluorescence spectral characteristic of different class phytoplankton is obtained in conjunction with phytoplankton dynamic processes composition, in conjunction with technical merit and the optical filter processing technology of current single color LED, select 412nm, 435nm, 455nm, 470nm, 507nm, 535nm, 567nm, 624nm, 635nm, 665nm etc. 10 excite wave band, select 620nm, 654nm, 670nm, 683nm, 692nm, 6 emission band such as 725nm, build the discrete three-dimensional fluorescence spectrum be made up of 36 characteristic spectrum points,
(2) then, take chlorophyll-a concentration as unified standard, by characteristic fluorescence spectrum to chlorophyll-a concentration normalization, set up different class planktonic algae normal scatter three-dimensional fluorescence spectrum storehouse;
(3) three-dimensional fluorescence spectrum discrete to testing sample characteristic spectrum the most similar or characteristic spectrum is found to combine in phytoplankton normal scatter three-dimensional fluorescence spectrum storehouse, be converted into linear regression problem again, namely carry out matching with each characteristic spectrum in normal scatter three-dimensional fluorescence spectrum storehouse to sample characteristic spectrum, concrete computation process is as follows:
1) the normal scatter three-dimensional fluorescence spectrum storehouse needing the phytoplankton of measuring is set up;
2) the discrete three-dimensional fluorescence spectrum of mixing phytoplankton sample is measured;
3) with each class phytoplankton normal scatter three-dimensional fluorescence spectrum for independent variable, mixing phytoplankton fluorescent spectrum is that dependent variable carries out multiple linear regression analysis, draws each class phytoplankton chlorophyll-a concentration.
Based on a phytoplankton identification determinator for discrete three-dimensional fluorescence spectrum, it is characterized in that: include excitation-emission driving circuit, excitation-emission optical system, fluorescence signal testing circuit, data acquisition and control circuit, described excitation-emission driving circuit includes digital regulation resistance, motor driver, and digital regulation resistance connects constant current source, analog switch, is connected with metal-oxide-semiconductor between digital regulation resistance and analog switch, described excitation-emission optical system includes columniform sample cell, bottom sample cell, 10mm place is equipped with the LED array of multistage ripple, low pass filter is placed between LED array and sample cell, photodetector is placed with bottom sample cell, be provided with fluorescence bands at the bottom of sample cell and between photodetector and select filter set, fluorescence bands selects filter set to be arranged on optical filter wheel, optical filter wheel is rotated by DC motor Driver, optical filter wheel is provided with photoelectricity pilot hole, optocoupler, each switch in parallel of analog switch and with each LED one_to_one corresponding control linkage of LED array, described fluorescence signal testing circuit includes the I/V change-over circuit, bandwidth-limited circuit, variable-gain amplification circuit, full-wave rectifying circuit, the low-pass filter circuit that link successively, and the signal output part of photodetector is connected with the signal input part of I/V change-over circuit, described data acquisition and control circuit include master controller, master controller is connected with data processing centre (DPC), analog to digital converter, the signal output part of data processing centre (DPC) is connected with display screen, the signal output part of low-pass filter circuit is connected with the signal input part of analog to digital converter, master controller and metal-oxide-semiconductor, analog switch, motor driver control linkage, under the control of master controller, excitation-emission driving circuit switches the LED array of multistage ripple by analog switch, band selection optical filter is switched by DC MOTOR CONTROL optical filter wheel, realize multiband timesharing to excite and receive with multiband fluorescence timesharing, fluorescence signal is converted into electric signal by photodetector, electric signal is amplified by fluorescence signal testing circuit nurses one's health into DC level, DC level signal converts data word signal to by analog to digital converter 44 and hands over data processing centre's analyzing and processing, the final discrete three-dimensional fluorescence spectrum measuring acquisition sample, and inverting obtains phytoplankton species and content in sample, measurement result is exported by display screen.
Described a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum, is characterized in that: described sample cell adopts diameter to be 10mm standard cylindrical quartz sample pool.
Described a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum, it is characterized in that: described LED array is made up of 10 LED altogether, excite the centre wavelength of LED to be respectively 412nm, 435nm, 455nm, 470nm, 507nm, 535nm, 567nm, 624nm, 635nm, 665nm.
Described a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum, it is characterized in that: a little higher than LED centre wavelength of low pass cutoff wavelength of the low pass filter placed between described LED array and sample cell, fluorescence bands light in filtering exciting light, reduces light source scattered light to the impact of fluorescence measurement.
Described a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum, is characterized in that: described photodetector adopts S3590-18 type PIN pipe, receives fluorescent with LED array direction in 90 °.
Described a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum, is characterized in that: described fluorescence bands selects the wave band of filter set to be respectively 620nm, 654nm, 670nm, 683nm, 692nm, 725nm.
Described a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum, is characterized in that: described excitation-emission optical system is arranged in optical texture shell, is covered with sun-shading cap above optical texture shell.
Advantages of the present invention is as follows:
(1) in order to distinguish and identify the phytoplankton of not fellow disciple, the present invention is according to the fingerprint characteristic of phytoplankton dynamic processes fluorescence spectrum, select different photosynthetic pigments characteristic of correspondence fluorescence peak as analytic target, propose a kind of phytoplankton identification based on discrete three-dimensional fluorescence spectrum and assay method, the inventive method farthest remains Phytoplankton Characteristic information in Three-Dimensional Fluorescence Spectra, has higher identification measure ability to phytoplankton.
(2) for phytoplankton fluorescence spectrum fingerprint characteristic, the present invention is using multiband LED array as excitation source, using multiband filter set as the band selector receiving fluorescence, devise a kind of discrete three-dimensional fluorescence spectrum determinator, this apparatus structure is simply compact, volume is little, easy to carry, be applicable to identification and the mensuration of on-the-spot phytoplankton.
Accompanying drawing explanation
Fig. 1 is phytoplankton discrete three-dimensional fluorescence spectrum point schematic diagram.
Fig. 2 is discrete three-dimensional fluorescence spectrum assay device structures schematic diagram.
Fig. 3 is excitation-emission driver' s timing figure.
Fig. 4 is excitation-emission optical system structure schematic diagram.
Embodiment
One, based on the phytoplankton identification assay method of discrete three-dimensional fluorescence spectrum
Phytoplankton living body fluorescent results from cell photosynthesis, closely related with Photosynthesis Pigment, and the exciting of fluorescence to depend on that with emission wavelength pigment is to the absorption of luminous energy, transmission and release characteristics.Due to the photosynthetic pigments composition of variety classes phytoplankton and content different, so photosynthetic pigments fluorescence has significant phytoplankton species finger print information, be the mark of desirable phytoplankton identification mensuration.
1, first the continuous three-dimensional fluorescence spectrum of the different class phytoplanktons of fluorescence spectrophotometer measurement is utilized, the fluorescence spectral characteristic of different class phytoplankton is obtained in conjunction with phytoplankton dynamic processes composition, in conjunction with technical merit and the optical filter processing technology of current single color LED, the present invention selects 412nm, 435nm, 455nm, 470nm, 507nm, 535nm, 567nm, 624nm, 635nm, 665nm etc. 10 excite wave band, select 620nm, 654nm, 670nm, 683nm, 692nm, 6 emission band such as 725nm, build the discrete three-dimensional fluorescence spectrum be made up of 36 characteristic spectrum points, as shown in Figure 1.
2, then, take chlorophyll-a concentration as unified standard, by characteristic fluorescence spectrum to chlorophyll-a concentration normalization, set up different class planktonic algae normal scatter three-dimensional fluorescence spectrum storehouse.
3, the basic thought based on the phytoplankton identification assay method of discrete three-dimensional fluorescence spectrum finds three-dimensional fluorescence spectrum discrete to testing sample characteristic spectrum the most similar or characteristic spectrum to combine, linear regression problem can be converted into, namely carry out matching with each characteristic spectrum in normal scatter three-dimensional fluorescence spectrum storehouse to sample characteristic spectrum, concrete computation process is as follows:
1) the normal scatter three-dimensional fluorescence spectrum storehouse needing the phytoplankton of measuring is set up;
2) the discrete three-dimensional fluorescence spectrum of mixing phytoplankton sample is measured;
3) with each class phytoplankton normal scatter three-dimensional fluorescence spectrum for independent variable, mixing phytoplankton fluorescent spectrum is that dependent variable carries out multiple linear regression analysis, draws each class phytoplankton chlorophyll-a concentration.
Two, based on the phytoplankton identification determinator of discrete three-dimensional fluorescence spectrum
According to the fingerprint characteristic of phytoplankton fluorescence spectrum, the present invention devises a kind of discrete three-dimensional fluorescence spectrum determinator, and apparatus structure as shown in Figure 2.Device forms primarily of four parts: excitation-emission driving circuit 10, excitation-emission optical system 20, fluorescence signal testing circuit 30, data acquisition and control circuit 40.Under the control of master controller 41, excitation-emission driving circuit by analog switch 13 switch multistage ripple LED array 21, control optical filter wheel 26 by direct current generator 25 and switch band selection optical filter 24, realize multiband timesharing to excite and multiband fluorescence timesharing reception, driver' s timing as shown in Figure 3.Fluorescence signal is converted into electric signal by photodetector 27, electric signal is amplified by fluorescence signal testing circuit 30 nurses one's health into DC level, DC level signal converts data word signal to by analog to digital converter 44 and hands over data processing centre (DPC) 42 analyzing and processing, the final discrete three-dimensional fluorescence spectrum measuring acquisition sample, and inverting obtains phytoplankton species and content in sample, measurement result is exported by display screen 43.
In Fig. 2: excitation-emission driving circuit 10 includes constant current source 11, analog switch 12, digital regulation resistance 13, metal-oxide-semiconductor 14, motor driver 15, excitation-emission optical system 20 includes LED array 21, low pass filter 22, sample cell 23, fluorescence bands selects filter set 24, direct current generator 25, optical filter wheel 26, photoelectricity pilot hole 27, optocoupler 28, photodetector 29, fluorescence signal testing circuit 30 includes I/V change-over circuit 31, bandwidth-limited circuit 32, variable-gain amplification circuit 33, full-wave rectifying circuit 34, low-pass filter circuit 35, data acquisition and control circuit 40 include master controller 41, data processing centre (DPC) 42, display screen 43 analog to digital converter 44.
As shown in Figure 4, optical system center is diameter 10mm standard cylindrical quartz sample pool 23 to excitation-emission optical system 20; Excitation source is made up of 10 LED altogether, 10mm place bottom distance sample cell, evenly be centered around around sample cell, the centre wavelength of LED is excited to be respectively 412nm, 435nm, 455nm, 470nm, 507nm, 535nm, 567nm, 624nm, 635nm, 665nm, low pass filter 22 is placed between LED and sample cell 23, the a little higher than LED centre wavelength of low pass cutoff wavelength, fluorescence bands light in filtering exciting light, reduces light source scattered light to the impact of fluorescence measurement; Photodetector 29 adopts S3590-18 type PIN to manage, be placed on bottom sample cell, become 90 directions to receive fluorescent with excitation source, fluorescence bands selects filter set 24 at the bottom of sample cell and between detector, wave band is respectively 620nm, 654nm, 670nm, 683nm, 692nm, 725nm, be arranged on optical filter wheel 26, coordinated by direct current generator 25, photoelectricity pilot hole 27 and optocoupler 28 and realize optical filter switching; Optical texture shell 2 combines with sun-shading cap 1 and realizes sample chamber, excitation-emission optical system has carried out optical encapsulant design, the fluorescent signals of phytoplankton is easily by extraneous illumination effect, for phytoplankton fluorescence spectral characteristic, adopt excitation light emission unit and the fluorescence detecting unit design of discretize wavelength, abandon the beam splitting system of conventional three-dimensional fluorescence spectrophotometer complexity, greatly simplified optical construction, reduce device volume and power consumption.
Excitation-emission driving circuit 10 selects the switching of filter set 24 for the excitation source LED array 21 and fluorescence bands controlling excitation-emission optical system 20.Excitation source driving circuit adopts constant current source type of drive, improves the stability of light source, uses many grades of digital regulation resistances 13 to regulate the drive current of each paths of LEDs, allow each paths of LEDs light intensity all at 1500 μm of ol/m
2about/s, ensures that phytoplankton active somatic cell can normal photosynthesis under being excited; Analog switch 12 is used to control the switching of 10 paths of LEDs; Use metal-oxide-semiconductor switch 14 pairs of light sources to carry out 10kHz modulation, allow the fluorescence exciting generation carry specific frequency information, to improve the signal to noise ratio (S/N ratio) that fluorescence signal is measured.Optical filter wheel driving circuit uses optocoupler to detect the position of optical filter wheel photoelectricity pilot hole determination optical filter wheel, is rotated, switch fluorescence bands and select optical filter by DC MOTOR CONTROL optical filter wheel.The all Control timing sequence of excitation-emission driving circuit are provided by master controller 41.
Fluorescence signal testing circuit 30 is made up of I/V change-over circuit 31, bandwidth-limited circuit 32, variable-gain amplification circuit 33, full-wave rectifying circuit 34 and low-pass filter circuit 35.I/V change-over circuit uses low noise, high-precision amplifier OPA637 carries out circuit voltage and transforms amplification, translated resistance 200k Ω; Bandwidth-limited circuit uses 4 rank Butterworth filtering circuits, and band passband rate 10kHz, carries out frequency-selecting output to signal; Variable-gain amplification circuit has 100,300,900,2400 fourth gear enlargement factors, and circuit regulates enlargement factor by the gain resistor of analog switch selective amplifier, and analog switch is controlled by master controller 41; Full-wave rectifying circuit and low-pass filter circuit composition demodulator circuit, nurse one's health into direct current signal by 10kHz sinusoidal signal, direct current signal size direct ratio and sinusoidal signal amplitude.
Data acquisition and control circuit 40 are made up of master controller 41, analog to digital converter 44, data processing centre (DPC) 42, display screen 43.The sequential of the driving of main controller controls excitation-emission and fluoroscopic examination circuit, receives the data-signal gathered by analog to digital converter, measures the discrete three-dimensional fluorescence spectrum obtaining sample; Data processing centre (DPC) utilizes the phytoplankton normal scatter three-dimensional fluorescence spectrum prestored to carry out matching to the fluorescent spectrum measured, and inverting obtains the Chlorophyll-a Content of different class phytoplankton; Sample variation three-dimensional fluorescence spectrum and concentration result of calculation are finally by display screen display translation.
Embodiment
Select the microcystic aeruginosa (Microcystisaeruginosa) of Cyanophyta, the chlorella (Chlorellavulgaris) of Chlorophyta, the mast algae (Fragilariasp) of Bacillariophyta, the light dinoflagellate (Glenodiniumgymnodinium) of Pyrrophyta and the avette hidden algae (Cryptomonasovata) of Cryptophyta test algae kind, get above-mentioned five kinds of algae purebred cultivation body of 15 days, use each purebred cultivation body chlorophyll-a concentration of high-efficient liquid phase chromatogram technique measuring, by purebred cultivation body using distilled water diluting certain multiple as sample mother liquor, sample mother liquor chlorophyll-a concentration is calculated according to dilution ratio, mother liquor chlorophyll-a concentration controls in 80-100 μ g/L scope.Then, with distilled water, five kinds of algae mother liquors are half-and-half diluted to respectively the purebred sample of 3 concentration gradients, then are hybridly prepared into 26 kinds of planktonic algae biased samples by volume ratio shown in table 2.
The volume ratio of planktonic algae biased sample prepared by table 1
Table 1 note: numeral 0 represents distilled water, and alphabetical A, B, C, D, E represent microcystic aeruginosa, chlorella, mast algae, light dinoflagellate and avette hidden algae respectively.
After the planktonic algae biased sample dark adatpation 30min of preparation, the planktonic algae identification determinator utilizing this problem to develop measures the discrete three-dimensional fluorescence spectrum of above-mentioned each sample, and calculate the chlorophyll-a concentration of each class planktonic algae of compound sample kind, each sample measures 3 times, mean value as measurement result, by the measurement result of fluorescence method and high-efficient liquid phase chromatogram technique measuring results contrast.The statistical study display of result: to purebred algae sample, fluorescence method measurement result is accurately high, closely, the relative error of microcystic aeruginosa, chlorella, mast algae, light dinoflagellate and avette hidden concentration of algae measurement result is respectively 0.7%, 7.64%, 5.73%, 4.66%, 8.68% for measured value and HPLC measured value; In biased sample, to concentration rate higher than 50% advantage algae, fluorescence method concentration measurement and HPLC measured value close, the relative error of microcystic aeruginosa, chlorella, mast algae, light dinoflagellate and avette hidden concentration of algae measurement result is respectively 6.42%, 10.26%, 5.78%, 4.66%, 9.84%.
Claims (8)
1., based on a phytoplankton identification assay method for discrete three-dimensional fluorescence spectrum, it is characterized in that, specifically comprise the following steps:
(1) the continuous three-dimensional fluorescence spectrum of the different class phytoplanktons of fluorescence spectrophotometer measurement is first utilized, the fluorescence spectral characteristic of different class phytoplankton is obtained in conjunction with phytoplankton dynamic processes composition, in conjunction with technical merit and the optical filter processing technology of current single color LED, select 10 excitation wavelength: 412nm, 435nm, 455nm, 470nm, 507nm, 535nm, 567nm, 624nm, 635nm, 665nm, select 6 emission wavelength: 620nm, 654nm, 670nm, 683nm, 692nm, 725nm, build the discrete three-dimensional fluorescence spectrum be made up of 36 characteristic spectrum points,
(2) then, take chlorophyll-a concentration as unified standard, by characteristic fluorescence spectrum to chlorophyll-a concentration normalization, set up different class planktonic algae normal scatter three-dimensional fluorescence spectrum storehouse;
(3) in phytoplankton normal scatter three-dimensional fluorescence spectrum storehouse, three-dimensional fluorescence spectrum discrete to testing sample characteristic spectrum the most similar or characteristic spectrum is found to combine, be converted into linear regression problem again, namely carry out matching with each characteristic spectrum in normal scatter three-dimensional fluorescence spectrum storehouse to sample characteristic spectrum, concrete computation process is as follows:
1) the normal scatter three-dimensional fluorescence spectrum storehouse needing the phytoplankton of measuring is set up;
2) the discrete three-dimensional fluorescence spectrum of mixing phytoplankton sample is measured;
3) with each class phytoplankton normal scatter three-dimensional fluorescence spectrum for independent variable, mixing phytoplankton fluorescent spectrum is that dependent variable carries out multiple linear regression analysis, draws each class phytoplankton chlorophyll-a concentration.
2. based on a phytoplankton identification determinator for discrete three-dimensional fluorescence spectrum, it is characterized in that: include excitation-emission driving circuit, excitation-emission optical system, fluorescence signal testing circuit, data acquisition and control circuit, described excitation-emission driving circuit includes digital regulation resistance, motor driver, and digital regulation resistance connects constant current source, analog switch, is connected with metal-oxide-semiconductor between digital regulation resistance and analog switch, described excitation-emission optical system includes columniform sample cell, bottom sample cell, 10mm place is equipped with the LED array of multi-wavelength, low pass filter is placed between LED array and sample cell, photodetector is placed with bottom sample cell, be provided with fluorescence bands at the bottom of sample cell and between photodetector and select filter set, fluorescence bands selects filter set to be arranged on optical filter wheel, optical filter wheel is rotated by DC motor Driver, optical filter wheel is provided with photoelectricity pilot hole, optocoupler, each switch in parallel of analog switch and with each LED one_to_one corresponding control linkage of LED array, described fluorescence signal testing circuit includes the I/V change-over circuit, bandwidth-limited circuit, variable-gain amplification circuit, full-wave rectifying circuit, the low-pass filter circuit that connect successively, and the signal output part of photodetector is connected with the signal input part of I/V change-over circuit, described data acquisition and control circuit include master controller, master controller is connected with data processing centre (DPC), analog to digital converter, the signal output part of data processing centre (DPC) is connected with display screen, the signal output part of low-pass filter circuit is connected with the signal input part of analog to digital converter, master controller and metal-oxide-semiconductor, analog switch, motor driver control linkage, under the control of master controller, excitation-emission driving circuit switches the LED array of multistage ripple by analog switch, band selection optical filter is switched by DC MOTOR CONTROL optical filter wheel, realize multiband timesharing to excite and receive with multiband fluorescence timesharing, fluorescence signal is converted into electric signal by photodetector, electric signal is amplified by fluorescence signal testing circuit nurses one's health into DC level, DC level signal converts digital signal to by analog to digital converter and hands over data processing centre's analyzing and processing, the discrete three-dimensional fluorescence spectrum of final acquisition sample, and inverting obtains phytoplankton species and content in sample, measurement result is exported by display screen.
3. a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum according to claim 2, is characterized in that: described sample cell employing diameter is the standard cylindrical quartz sample pool of 10mm.
4. a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum according to claim 2, it is characterized in that: described LED array is made up of 10 LED altogether, excite the centre wavelength of LED to be respectively 412nm, 435nm, 455nm, 470nm, 507nm, 535nm, 567nm, 624nm, 635nm, 665nm.
5. a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum according to claim 2 or 4, is characterized in that: a little higher than LED centre wavelength of low pass cutoff wavelength of the low pass filter placed between described LED array and sample cell.
6. a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum according to claim 2, is characterized in that: described photodetector adopts S3590-18 type PIN pipe, receives fluorescent with LED array direction in 90 °.
7. a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum according to claim 2, is characterized in that: what described fluorescence bands selected filter set is respectively 620nm, 654nm, 670nm, 683nm, 692nm, 725nm through wavelength.
8. a kind of phytoplankton identification determinator based on discrete three-dimensional fluorescence spectrum according to claim 2, is characterized in that: described excitation-emission optical system is arranged in optical texture shell, is covered with sun-shading cap above optical texture shell.
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| JP2006284335A (en) * | 2005-03-31 | 2006-10-19 | Univ Nagoya | Chlorophyll fluorescence measuring method and chlorophyll fluorescence measuring device |
| CN1696663A (en) * | 2005-05-20 | 2005-11-16 | 中国海洋大学 | Method for quick distinguishing and determining phytoplankton in red tide |
| CN101561395B (en) * | 2009-03-20 | 2012-05-23 | 中国海洋大学 | Phytoplankton composition quick determination method |
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