CN208297348U - Seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus - Google Patents
Seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus Download PDFInfo
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- CN208297348U CN208297348U CN201820051533.6U CN201820051533U CN208297348U CN 208297348 U CN208297348 U CN 208297348U CN 201820051533 U CN201820051533 U CN 201820051533U CN 208297348 U CN208297348 U CN 208297348U
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- phycocyanin
- phycoerythrin
- seawater
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- chlorophyll
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Abstract
The utility model belongs to Portable seawater fluorescence in situ detection technology field, disclose seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus and method, including minitype piezoelectric pump, flow cell, excitation light source, lens, optical splitter, filter and photodiode, excitation light source can excite flow cell.The technical program uses microfluidic chip technology, will pre-process, and reacts, the units such as separation and monitoring are integrated into chip, the measurement of seawater chlorophyll a, phycocyanin, phycoerythrin concentration, automatic sampling, analysis, correction, high degree of automation are rapidly completed on chip, it is high-efficient, equipment instrument reduces optimization, and level of integrated system is high, low in energy consumption, it is at low cost, it is suitble in situ measurement, rapid survey, continuous measurement, laboratory measurement and Portable field measurement.
Description
Technical field
The invention belongs to Portable seawater fluorescence in situ detection technology field, and in particular to a kind of based on micro-fluidic chip
Seawater chlorophyll a, phycocyanin and phycoerythrin three-parameter measuring device.
Background technique
In the prior art, the seawater chlorophyll a based on chlorophyll fluorescence techniques, phycocyanin, phycoerythrin three parameter
Measuring device and method not yet making breakthroughs property are in progress, and tri- parameter of research Multi-Color-PAM, Trilux of related fields is glimmering
Optical sensor etc. foreign countries instrument can satisfy needs, however the quasi-instrument is more complicated, expensive, sensitivity is low cannot
It is advantageously applied in live seawater.Moreover, the research of the photosynthesis measurement method based on fluorescent technique is mostly all with fresh water
Blue, green alga is research object, is not appropriate for the research of marine algae.Traditional instrument and measurement method time of measuring are long, and operation is multiple
It is miscellaneous, error is big, power consumption is big, volume is big, at high cost, and can not long-term on-line measurement.Marine field not yet occurs green based on leaf
The research of the chlorophyll a, phycocyanin, phycoerythrin three-parameter measuring method of plain fluorescent technique.Therefore it provides a kind of efficiently low
Portable ocean chlorophyll a cost, in situ, phycocyanin, phycoerythrin three-parameter measuring device and method are necessary
's.
Summary of the invention
It is a kind of based on micro-fluidic chip measurement seawater chlorophyll it is an object of the invention in view of the above technical problems, provide
A, phycocyanin, phycoerythrin concentration device and method.To achieve the above object, using following technical scheme:
Seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus include sampling apparatus and detection device, sampling
Device includes the micro-fluidic chip with circulation structure, filtering head, seawater sample introduction pipeline, waste discharge pipeline, miniature electromagnetic valve, miniature
Air pump, micro-injection pump, flow cell；Detection device includes the excitation LED light of tri- kinds of different-wavebands of 470nm, 610nm, 530nm
Source, optical fiber, photodiode, lens, optical filter and signal processing unit.It is gentle that micro-fluidic chip sampling apparatus is equipped with liquid
The chip channel of body loop, filtering head and chip directly pass through the seawater sample introduction pipeline connection, and chip channel is equipped with multiple
The miniature electromagnetic valve of control channel on-off is additionally provided with miniature voltage pump, flow cell and recoil channel in chip channel.Detection device
In the excitation LED of three kinds of different wave lengths pass through optical fiber respectively and be connected to lens and optical filter, light splitting is equipped in front of lens and optical filter
Mirror, spectroscope front are connected to measurement photodiode by optical fiber and compare photodiode, and micro-fluidic chip is additionally provided with waste discharge
Pipeline and signal processing unit, signal processing unit are used based on the inspection synchronous with orthogonal digital of programmable-gain trans-impedance amplifier
The light path detections such as wave energy circuit composition, the circuit of the special research and development can effectively eliminate environment light, different frequency noise
Influence to measurement result.
Device carries out the inspection of three paths of LEDs optical paths after seawater after filtering is injected into flow cell one loop of formation by microchannel
It surveys, the LED light source of three kinds of different wave lengths is used to realize the photosynthesis measurement of Phytoplankton & Suspension as excitation light source.It can be improved measurement
Precision and accuracy, integrated level is high, and replaceability is strong, portable to be used for laboratory, the station and buoy in situ detection simultaneously.Swash
Light emitting source LED is replaceable device, can be replaced according to different measurement requests or to excitation light source LED.Device carries
The circuit of special development can effectively shield influence of the external environmental light to measurement result.
Filtering head is the bag type filter that aperture is not more than 10um；
Sample introduction pipeline, waste discharge pipeline and backflushing pipe internal diameter are 0.5mm-1mm, outer diameter 1mm-2mm；
Flow cell detection window internal diameter is 1mm-3mm, and circulation pond length is 30-50mm；
Spectroscope prevents LED because the luminous intensity that works long hours is unstable, wherein LED beam is as exciting light all the way, separately
Light beam compensates dynamic excitation light source as LED reference light all the way outside；
Respectively as fluorescence detector and reference light source receiver, what photodiode was all made of is two-way photodiode
Shore pine, noise is in 2pA or less；
The flow velocity of minitype piezoelectric pump and micro- air pump is 50-500ul/min；
Optical filter, lens, spectroscope pass through optical fiber and are integrated in micro-fluidic chip.
Seawater chlorophyll a, phycoerythrin, three parameter of phycocyanin measurement method be the sea based on chlorophyll fluorescence techniques
Water by micro-fluidic chip enter is integrated in the flow cell of chip interior, recycle be integrated on chip 470nm, 610nm,
The LED excitation light source of tri- kinds of different wave lengths of 530nm irradiates phytoplankton suspension, the fluorescence volume that measurement phytoplankton issues respectively
Sub- intensity to realize seawater chlorophyll a, phycocyanin, phycoerythrin concentration real-time measurement.Method includes the following steps:
1) valve 2,4,6,8,9 and minitype piezoelectric pump 1 are opened, other pump valves is closed, enters seawater after filtering head B
Chip channel carries out cleaning and being full of seawater sample introduction pipeline C and flow cell M, and extra seawater is discharged by waste discharge pipeline D；
2) valve 2,4,6,8,9 and minitype piezoelectric pump 1 are closed, realizes that the LED of three road different wave lengths swashs by analog switch
The switching of light emitting source, sense photodiode N and comparison photodiode O are detected irradiate in three kinds of different wave length exciting lights respectively
The beam intensity ratio of fluorescence and reference light, calculation formula are in lower flow cellWherein I is the fluorescence intensity of excitation, I0For
Exciting light luminous intensity；After chlorophyll a, phycocyanin and phycoerythrin in seawater are excited using LED light, difference can be issued
The fluorescence signal of wavelength, the fluorescence intensity issued are as follows:
F=k φ I0(1-e-εcb)
Wherein k is instrumental constant, and Φ is substance fluorescence efficiency, I0For exciting light luminous intensity, c is material concentration, and b is sample
Product optical path difference, ε are molar absorption coefficient.
Find out from formula, when exciting light luminous intensity is stablized, fluorescence intensity only with chlorophyll a, phycocyanin, phycoerythrin
Concentration is related, the relationship of chlorophyll concentration and fluorescence intensity F are as follows: in c=A-Blg (D-F) formula
D=kQI0.The intensity F for detecting the fluorescence signal received accordingly can be detected seawater chlorophyll a, phycoerythrin, algae indigo plant egg
White concentration c.
The technical program uses microfluidic chip technology, will pre-process, and reacts, and the units such as separation and monitoring are integrated into chip
It is interior, the measurement of seawater chlorophyll a, phycocyanin, phycoerythrin concentration, automatic sampling, analysis, school are rapidly completed on chip
Just, high degree of automation, high-efficient, equipment instrument reduces optimization, and level of integrated system is high, low in energy consumption, at low cost, is suitble in situ survey
Amount, rapid survey, continuous measurement, laboratory measurement and Portable field measurement；It is used under the LED exciting light irradiation of different wave length
Variable gain amplifier and digital quadrature synchronization detecting circuit and double photo paths scheme, effectively eliminate environment light and LED current
Change influence of the noise to measurement result caused by different frequency, improves the detection limit and detection accuracy of device；Backwash system
System effectively cleaning filtering head and sample introduction pipeline and circulation pipeline, extend instrument original position runing time；Pure optical means is not necessarily to chemistry
Reagent, it is not damaged to organism.
Detailed description of the invention
Fig. 1: the structural schematic diagram of three-parameter measuring device of the invention.
Wherein: 1. minitype piezoelectric pumps；2-9. valve；10. micro- air pump；A. micro-fluidic chip；B. filtering head；C. sample introduction pipeline；
D. waste discharge pipeline；E. backflushing pipe；F, G, H. excitation light source；I. optical fiber；J. optical filter；K. spectroscope；L. lens；M. it circulates
Pond；N. photodiode；O. photodiode is compared；P. detection circuit.
Specific embodiment
As shown in Figure 1, being done by alga classifying measuring device and method of the embodiment to the technical program micro-fluidic chip
Further instruction.
Using China coastal seas representativeness algae as research object, with the excitation of tri- kinds of different wave lengths of 470nm, 610nm, 530nm
Light source, the fluorescence signal generated respectively to excitation detect, and are joined to realize with chlorophyll a, phycocyanin, phycoerythrin three
Several measurements.
Detection device includes the micro-fluidic chip A with loop structure, filtering head B, seawater sample introduction pipeline C, waste discharge pipeline
D, high brightness excitation LED light F, G, H, optical fiber I, optical filter J, spectroscope K, the lens L, stream of backflushing pipe E, three kinds of different-wavebands
Logical pond M, measurement photodiode N, reference photodiode O and detection circuit P.Wherein micro-fluidic chip A is equipped with multiple liquid
The chip channel of body loop, the chip channel are equipped with the miniature electromagnetic valve 2-9 of multiple control chip channel switches, and chip is logical
Minitype piezoelectric pump 1, flow cell M, the flow cell M are additionally provided on road by optical fiber I with lens L, optical filter J, with reference to photoelectricity two
Pole pipe N, measurement photodiode O are connected.
Filtering head B is mainly the filter device of the metal sintered material of 10um, mainly filters the particulate matter in seawater
Fall so as not to particulate matter blocking pipeline and influence measurement result.The filtering head B is placed in seawater (in situ measurement).Filtering head B
One end is connected by valve 4 with sample introduction pipeline C, and the other end is connected by valve 2 with minitype piezoelectric pump 1, and third section connection valve 3 carries out
Emptying recoil.
Minitype piezoelectric pump 1 is mainly sent filtered seawater to carrying out subsequent inspection in flow cell M by sample introduction pipeline C
It surveys, flow cell M will be discharged full of seawater extra after sample introduction product by waste discharge pipeline D.
Micro- air pump 10 is connected by valve 7, valve 5, valve 4 with filtering head B respectively, mainly flushing-back filtering head, prevents biology attached
, extend the on-line monitoring time.
Excitation light source F, G and H of three kinds of different-wavebands pass through optical fiber I respectively and are connected with spectroscope K, and spectroscope K will be excited
Light source is divided into two-way, is connected by optical fiber I with flow cell M after being connected all the way by optical fiber I with lens filter set, as excitation
Light excites algae in seawater to generate fluorescence signal.In addition it is connected all the way as reference light with reference photodiode O.Reference is main
Effect is small in seawater inside detection error brought by elimination high-brightness LED lamp luminous intensity variations and flow cell
The detection bring of seawater fluorescence signal is interfered caused by particle.
Detection circuit is the circuit based on programmable-gain Yu quadrature synchronization detection function of special research and development, can effectively be disappeared
Except the variation of environment light and excitation light source program the influence generated to measurement result.
It is as follows to seawater measurement key step:
1. opening valve 2,4,6,8,9 and minitype piezoelectric pump 1, closes other pump valves, enters filtered seawater sample
To flow cell, and it is full of flow cell M, extra seawater is flowed out by waste discharge pipeline D.
2. being separately turned on the highlighted excitation LED light of 470nm, 610nm and 530nm, measures swash under three kinds of different wave lengths respectively
The intensity of the fluorescence of hair, and fluorescent value is calculated, bring the fluorescent value being calculated into formula respectively to obtain in three kinds of seawater
The concentration value of algae.
3. on-site measurement seawater sample is respectively in the excitation fluorescence that excitation wavelength is at 470nm, 610nm and 530nm wavelength
Value can obtain the concentration value of three kinds of different algal species in seawater.
4. measurement after the completion of, close valve 2,6,8,9 after, open valve 3,4,5,7, starting micro air pump 10 to filtering head B into
Row recoil, to extend the service life of filtering head.
Embodiment only illustrates technical solution of the present invention, rather than carries out any restrictions to it；Although with reference to the foregoing embodiments
Invention is explained in detail, for those of ordinary skill in the art, still can be to previous embodiment institute
The technical solution of record is modified or equivalent replacement of some of the technical features；And these modifications or substitutions, and
The essence of corresponding technical solution is not set to be detached from the spirit and scope of claimed technical solution of the invention.
Claims (7)
1. seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus, it is characterised in that: including minitype piezoelectric pump, stream
Logical pond, excitation light source, lens, optical splitter, filter and photodiode, excitation light source can excite flow cell, excitation light source and
Flow cell is connect with lens respectively by optical fiber, and lens are connect by optical fiber with spectroscope, and spectroscope passes through optical fiber and photoelectricity two
By sample introduction piping connection, micro- air pump is connected to sample feeding pipe road for pole pipe connection, minitype piezoelectric pump and flow cell, forms recoil tube
Road.
2. seawater chlorophyll a according to claim 1, phycocyanin and phycoerythrin parameter measuring apparatus, feature exist
In: flow cell is additionally provided with waste discharge access.
3. seawater chlorophyll a according to claim 1 or 2, phycocyanin and phycoerythrin parameter measuring apparatus, feature
It is: further includes filtering head.
4. seawater chlorophyll a according to claim 3, phycocyanin and phycoerythrin parameter measuring apparatus, feature exist
In: filtering head is the bag type filter that aperture is not more than 10um.
5. seawater chlorophyll a according to claim 1 or 2, phycocyanin and phycoerythrin parameter measuring apparatus, feature
It is: further includes detection circuit.
6. seawater chlorophyll a according to claim 5, phycocyanin and phycoerythrin parameter measuring apparatus, feature exist
In: detection circuit is the optical path detection system based on programmable-gain trans-impedance amplifier Yu orthogonal digital synchronous detection functional circuit
System.
7. according to claim 1,2,4,6 any seawater chlorophyll as, phycocyanin and phycoerythrin parameter measurement dress
It sets, it is characterised in that: sample introduction pipeline, waste discharge pipeline and backflushing pipe internal diameter are 0.5mm-1mm, outer diameter 1mm-2mm.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108414477A (en) * | 2018-01-12 | 2018-08-17 | 山东省科学院海洋仪器仪表研究所 | Seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus and method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108414477A (en) * | 2018-01-12 | 2018-08-17 | 山东省科学院海洋仪器仪表研究所 | Seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus and method |
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Effective date of registration: 20211019 Address after: 266000 No.2 Huiying street, Shanghe demonstration zone, Jiaozhou City, Qingdao City, Shandong Province Patentee after: Qingdao Mihai Technology Co.,Ltd. Address before: 266000 Building 2, phase I, blue Silicon Valley entrepreneurship center, Qingdao blue Silicon Valley core area, aoshanwei street, Qingdao, Shandong Province Patentee before: INSTITUTE OF OCEANOGRAPHIC INSTRUMENTATION, SHANDONG ACADEMY OF SCIENCES |