CN102095686A - Method for detecting and analyzing blue-green algae - Google Patents
Method for detecting and analyzing blue-green algae Download PDFInfo
- Publication number
- CN102095686A CN102095686A CN 201010598251 CN201010598251A CN102095686A CN 102095686 A CN102095686 A CN 102095686A CN 201010598251 CN201010598251 CN 201010598251 CN 201010598251 A CN201010598251 A CN 201010598251A CN 102095686 A CN102095686 A CN 102095686A
- Authority
- CN
- China
- Prior art keywords
- blue
- green algae
- sample
- liquid
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000192700 Cyanobacteria Species 0.000 title claims abstract description 127
- 238000000034 method Methods 0.000 title claims abstract description 15
- 239000007788 liquid Substances 0.000 claims abstract description 55
- 238000001514 detection method Methods 0.000 claims abstract description 35
- 239000012530 fluid Substances 0.000 claims abstract description 23
- 238000004458 analytical method Methods 0.000 claims abstract description 20
- 230000003287 optical effect Effects 0.000 claims abstract description 20
- 238000012545 processing Methods 0.000 claims abstract description 20
- 238000007405 data analysis Methods 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000001914 filtration Methods 0.000 claims abstract description 8
- 238000006243 chemical reaction Methods 0.000 claims abstract description 5
- 239000000523 sample Substances 0.000 claims description 100
- 230000035479 physiological effects, processes and functions Effects 0.000 claims description 21
- 230000011664 signaling Effects 0.000 claims description 13
- 238000005259 measurement Methods 0.000 claims description 10
- 239000002245 particle Substances 0.000 claims description 7
- 239000004576 sand Substances 0.000 claims description 6
- 238000013459 approach Methods 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 4
- 230000003760 hair shine Effects 0.000 claims description 4
- 230000005693 optoelectronics Effects 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 3
- 239000012780 transparent material Substances 0.000 claims description 3
- 238000004904 shortening Methods 0.000 claims description 2
- 230000012010 growth Effects 0.000 abstract description 10
- 230000001678 irradiating effect Effects 0.000 abstract 1
- 239000003643 water by type Substances 0.000 description 13
- 108010053210 Phycocyanin Proteins 0.000 description 7
- 238000010586 diagram Methods 0.000 description 6
- 238000005286 illumination Methods 0.000 description 5
- 238000002795 fluorescence method Methods 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000005791 algae growth Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000012851 eutrophication Methods 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000000695 excitation spectrum Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 238000013316 zoning Methods 0.000 description 1
Images
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention relates to a method for detecting and analyzing blue-green algae, which comprises the following detection and analysis steps that: a sample adsorbing and liquid path control unit transfers two kinds of liquid into a flowing chamber, and sample liquid to be detected is coated with sheath fluid to flow through a detection region of the flowing chamber; a light source irradiates on the sample liquid to be detected which flows through the detection region of the flowing chamber; cells of blue-green algae in the sample liquid to be detected are irradiated by the irradiating light in the detection region to generate an optical signal, the optical signal is received by a photoelectric detector and is subjected to photoelectric conversion to form an electric signal, and the electric signal is sent to a signal processing and data analysis unit; and the signal processing and data analysis unit carries out filtering processing on the electric signal of the photoelectric detector and analyzes statistical data of a sample. By the method, the cells of blue-green algae in the sample to be detected can be measured one by one. The concentration of the blue-green algae can be accurately obtained, and different physiological periods of the blue-green algae can be analyzed. The method contributes to carrying out early warning on the growth condition of the blue-green algae in a water area.
Description
Technical field
The present invention relates to the check and analysis method of a kind of blue-green algae, be used for the different physiology phase of blue-green algae is analyzed, help the blue-green algae situation in the waters is given warning in advance.
Background technology
The lake is one of human most important water resource, also is China's population and industrial accumulation area on every side in the lake.Because the influence of mankind's activity, this year, the eutrophication in lake was on the rise, and Resource Properties such as Taihu Lake, big-and-middle-sized lake, Chaohu, Dian Chi, Dongting Lake, Hongchehu Lake, Baiyang Lake are on the hazard.The direct result of eutrophication is exactly the generation of blue-green alga bloom, and the blue-green alga bloom of Taihu Lake outburst in 2008 has had a strong impact on peripheral resident's normal life.How before the blue-green alga bloom outburst, to carry out early warning accurately and become extremely important.It is practicable approach that blue-green algae content in the water is carried out continuous monitoring.
Monitoring to blue-green algae all is to judge its concentration by detecting the absorption intensity or the fluorescence intensity of blue-green algae under specific wavelength at present, and its technology is:
1. utilize the phycocyanin in the blue-green algae that the absorption characteristic of the light of specific wavelength is carried out the absorbance log measurement to determine blue-green algae concentration, the quantitative test foundation is a Lambert-Beer's law, we are simply referred to as absorption process, as Fig. 1, comprise light source 1, sample cell 2, optical receiver 3, illumination beam 4, transmitted light beam 5.
2. utilize the fluorescence of phycocyanin specific wavelength of stimulated emission after absorbing light is shone, determine blue-green algae concentration, abbreviate fluorescence method as according to the size of detected fluorescence volume.The excitation wavelength peak value of phycocyanin is at 621nm, and the peak value of emission wavelength is at 646nm, is that the light source with this wave band shines mostly in the prior art.Fig. 2 has provided the synoptic diagram of fluorescence method, comprises light source 1, sample cell 2, and, illumination beam 6, fluorescent light beam 7, detector 8, collecting lens 9.Fig. 3 is another embodiment of fluorescence method, and ultimate principle is the same, all is the concentration of judging blue-green algae by the size of fluorescence intensity, comprises light source 1, sample cell 2, illumination beam 6, fluorescent light beam 7, detector 8, color filter 10.
The defective of said method is:
1. what detect is the overall of a sample, but not the individuality of each blue-green algae cell, intensity variations amount (absorbing light or fluorescence) can not be directly corresponding with blue-green algae concentration, need realize by the calibration system of complexity, conventional method is a sediments microscope inspection, waste time and energy very much, and same detecting instrument all needs to calibrate again to different waters.
2. because the blue-green algae fluorescence intensity of different physiology phases is different, can cause the inaccurate of concentration calibration.For example: the blue-green algae cell fluorescence amount in growth period is greater than the blue-green algae cell fluorescence amount in maturity stage, so low concentration growth period the blue-green algae cell and the maturity stage blue-green algae cell of high concentration just may obtain same fluorescence volume, this moment, calibration lost meaning.
3. the just concentration by the blue-green algae cell that detects can't judge that the early warning effect is relatively poor to the physiology phase of the blue-green algae in waters.
The best method that addresses the above problem is to adopt flow cytometer that sample to be tested is measured, but flow cytometer is bulky, heavy, cost an arm and a leg, and need outside machine, carry out sample process, only be adapted at lab investigation, can not carry out on-the-spot test and on-line real time monitoring.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, thereby the check and analysis method of a kind of blue-green algae is provided, this method is fit to the blue-green algae of in-site measurement to be analyzed, and can realize the contained blue-green algae cell of sample to be tested is measured one by one; Not only accurately obtain the concentration of blue-green algae, and the different physiology phase of blue-green algae is analyzed, help the blue algae growth situation in the waters is given warning in advance.
According to technical scheme provided by the invention, the check and analysis method of a kind of blue-green algae comprises following check and analysis step:
(1), sheath stream forms: measurand is formed by two kinds of liquid, and a kind of is sample to be tested liquid, at first gathers water sample with sample-sucking device, and particle diameter is greater than the sand grain of 1mm in the filtering water sample; Inner through screening deaeration and sand grains at sample-sucking device then, remaining phytoplankton is as sample to be tested liquid; Another kind is a sheath fluid; By inhaling sample and liquid road control module two kinds of liquid are delivered in the flow chamber, sample to be tested liquid is at the dirty via flow of the parcel of sheath fluid chamber detection zone, under the effect of fluid focus, sample liquid is compressed to the width of 2 blue-green algae cells, makes the blue-green algae cell one by one by the flow chamber detection zone;
(2), rayed part:
Light source forms an oval hot spot through first condenser lens or lens combination, and oval hot spot shines on the sample to be tested liquid that flows through in the flow chamber detection zone; Sheath fluid forms laminar flow in the flow chamber commutating zone, in the flow chamber accelerating region, sample liquid is focused on gradually, be sample flow until in the flow chamber detection zone, shortening sample to be tested hydraulic pressure into diameter less than 2 blue-green algae cell yardsticks, blue-green algae cell in the sample to be tested liquid just can only pass through the irradiation of detection zone irradiates light one by one like this, can only detect a cell in each detection incident;
(3), light-receiving probe portion: the blue-green algae cell one by one is subjected to the irradiation of detection zone irradiates light and produces optical signalling, and described optical signalling is collected by second condenser lens, enters photodetector by long logical color filter again; Optical signalling is received by photodetector, form electric impulse signal through after the opto-electronic conversion, deliver to signal Processing and data analysis unit, a stream of cells just forms an electric impulse signal when crossing detection zone, has so just realized the measurement one by one of individual cells in the sample;
(4) signal Processing and data analysis part: signal Processing and data analysis unit are carried out Filtering Processing to the electric signal of photodetector, and the statistics of sample analyzed, in the blue-green algae statistics of one dimension at least, divide at least 2 zones according to analytical approach, the different physiology phases of representing blue-green algae respectively, blue-green algae cell in the zone is counted respectively and analyzed, obtain the blue-green algae concentration of the blue-green algae total concentration of sample liquid, different physiology phases, the shared number percent of different physiology phase blue-green algaes; The total number that described blue-green algae total concentration is all pulses in the one-shot measurement process is divided by tested volume.
Described flow chamber comprises the guide hole that a flow of liquid is crossed, and this guide hole is divided into three zones: commutating zone, accelerating region, detection zone; Described detection zone is that the rayed zone is square hole or the rectangular opening that a length of side is 200um～400um, is made by optically transparent material; The part of the inlet flaring of described square hole or rectangular opening forms accelerating region, and the part that sheath fluid passage and sample liquid passage are nested is a commutating zone.
Described sample liquid channel outlet is the circular hole of diameter 0.3mm.
Described light source adopts light emitting diode; Described photodetector adopts photodiode.
Wavelength 620 ± the 5nm of described light emitting diode, printing opacity wavelength are 640nm～850nm.
The liquid stream of described flow chamber internal flow will satisfy laminar flow condition, i.e. Reynolds number＜2300; Sheath stream carries out fluid focus to sample flow, and sample flow is compressed to width less than 2 blue-green algae cell dias.
The length of described oval hot spot minor axis is that the length of 20 ± 5um, major axis is 200 ± 20um; The direction of oval hot spot minor axis is consistent with the cell flow direction, and oval hot spot long axis direction is perpendicular to cell flow direction and direction of beam propagation.
Described oval hot spot in the size requirements of flow chamber center is: less than 2 blue-green algae diameters, be not less than hole width in the flow chamber on the direction that flows perpendicular to beam propagation and sample at the same time on the sample flow direction; On perpendicular to the direction of beam propagation, light signal is collected on the photodetector with second condenser lens of a large-numerical aperture.
The optical signalling that produces after described blue-green algae cell is subjected to shining comprises: forward scattering light, side scattered light and fluorescence, described forward scattering light refer to the scattered light in 1 ~ 5 degree scope along the light path direction of propagation, have reflected the volume size of cell; Side scattered light refers to the scattered light perpendicular to optical propagation direction, and angular range is 75 degree ~ 115 degree, has reflected the inside complexity of cell; The angular range of fluorescence is consistent with the side scattered light angle, and the wavelength coverage of its fluorescence is 640nm ~ 850nm.
Advantage of the present invention is:
1. solve the calibration problem of traditional instrument.Blue-green algae cell number contained in the sample to be tested to designated volume is counted one by one, obtains blue-green algae concentration accurately, and need not obtain blue-green algae concentration by the scaling system of complexity, and measurement result is more accurate.
2. when measuring concentration, can analyze, the blue algae growth situation in waters is given warning in advance the different physiology phase of blue-green algae in the sample.
3. with the LED illumination, greatly reduce instrument cost and size, help the field condition operation.
Description of drawings
Fig. 1 is existing transmission beam method schematic diagram.
Fig. 2 is existing fluorescence method schematic diagram.
Fig. 3 is a blue-green algae fluorescence detecting sensor schematic diagram of using optical fiber.
Fig. 4 is the systematic schematic diagram that adopts the present invention to detect.
Fig. 5 is the structural drawing of photoelectric sensor of the present invention.
Fig. 6 (a) is the flow chamber sectional view.
Fig. 6 (b) is the A-A sectional view of Fig. 6 (a).
Fig. 7 is the detection synoptic diagram of detection zone in the flow chamber.
Fig. 8 is the spectral curve of color filter.
Fig. 9 is a blue-green algae cell statistic histogram.
Embodiment
The invention will be further described below in conjunction with drawings and Examples.
As Fig. 4, shown in Figure 5, check and analysis of the present invention system comprises photoelectric sensor 100, signal Processing and data analysis unit 200, inhales sample and liquid road control module 300; Described photoelectric sensor 100 comprises light irradiation unit 11, flow chamber 12 and light-receiving probe unit 13.Light-receiving probe unit 13 output terminals of described photoelectric sensor 100 are connected with data analysis unit 200 input ends with signal Processing, described suction sample and the control of liquid road
Shown in Fig. 6 (a), Fig. 6 (b), described flow chamber 12 is placed in the flow chamber seat, includes a guide hole that allows sheath fluid and sample liquid flow through, and this guide hole is divided into three zones: commutating zone 121, accelerating region 122, detection zone 123.Wherein detection zone 123, and just the rayed zone is square hole or the rectangular opening that a length of side is 200um～400um, and the profile of detection zone is a cube or rectangular parallelepiped, is made by optically transparent material.The inlet flaring of described square hole is the sheath fluid passage, parallel insertion sample liquid passage in the sheath fluid passage, sample liquid channel outlet is in described square hole inlet flaring zone, with the square hole inlet over against, described sample liquid channel outlet is the circular hole of diameter 0.3mm.The part of the inlet flaring of square hole forms accelerating region 122, and the part that sheath fluid passage and sample liquid passage are nested is a commutating zone 121.
Described light irradiation unit 11 comprises light source 1 and first condenser lens or lens combination 16, places the endoporus of light source assembly; The light beam scioptics 16 that light source sends form a focal beam spot and shine on the sample flow that flows through in the flow chamber; Described light irradiation unit 11 comprises a red LED at least as light source, and its wavelength is at 620nm ± 5nm;
3) light-receiving probe unit 13 comprises second condenser lens 7, long logical color filter 10 and photodetector 8, and described second condenser lens 7 is placed in the lens mount; Logical color filter 10 of described length and photodetector 8 are placed in the probe assembly, and the light signal that produces after the particle in the sample flow is subjected to shining is collected on the detector, and carries out opto-electronic conversion.
The check and analysis method of a kind of blue-green algae of the present invention may further comprise the steps:
1, sheath stream forms: measurand is formed by two kinds of liquid.The one, sample to be tested liquid is at first gathered water sample with sample-sucking device, and particle diameter is greater than the sand grain of 1mm in the filtering water sample; Inner through screening deaeration and sand grains at sample-sucking device then, remaining phytoplankton is as sample to be tested liquid; Another kind is a sheath fluid, by inhaling sample and liquid road control module 300 two kinds of liquid is delivered in the flow chamber 12, and two kinds of liquid flow through in flow chamber 12, and sample to be tested liquid is at the dirty via flow of the parcel of sheath fluid chamber detection zone 123.
The fluid of described flow chamber 12 internal flows will satisfy laminar flow condition, i.e. Reynolds number＜2300.Sheath stream carries out fluid focus to sample flow, and sample flow is compressed to width less than 2 blue-green algae cells, so that blue-green algae cell passing through one by one.
The sheath fluid that relates among the present invention is a physiological saline.Sample liquid promptly is filtered the back, contains the lake water of tested blue-green algae cell.
Described suction sample and liquid road control module 300 comprise: pressure source, gas, liquid pipeline, valve, control circuit, sample filtering device.Pressure source is air pump or syringe.
2, rayed: light source 1 adopts light emitting diode, through first condenser lens or lens combination 16, form an oval hot spot 14, oval hot spot shines on the sample to be tested liquid that flows through in the flow chamber detection zone 123, when the blue-green algae cell through above-mentioned fluid focus passed through one by one, they will be one by one illuminated;
3, light-receiving probe portion: blue-green algae cell 15 is by the irradiation of detection zone 123 irradiates lights, through producing optical signalling after the irradiation, described optical signalling promptly by second condenser lens 7, enters photodetector 8 by long logical color filter 10 by light-receiving probe unit 13 again; Optical signalling is received by photodetector 8, form electric impulse signal through after the opto-electronic conversion, deliver to signal Processing and data analysis unit 200, each cell will produce the one or more one to one electric impulse signals of cell characteristic therewith when passing through, so just realized the measurement of individual cells in the sample;
Described blue-green algae cell 15 is subjected to shining the back and produces optical signalling, and optical signalling comprises: forward scattering light, side scattered light and fluorescence.Forward scattering light FSC refers to the scattered light in 1 ~ 5 degree (± 0.3) scope along the light path direction of propagation, has reflected the volume size of cell.Side scattered light SSC refers to the scattered light perpendicular to optical propagation direction, and angular range is 75 degree ~ 115 degree (± 5 degree), has reflected the inside complexity of cell.The angular range of fluorescence is consistent with the side scattered light angle, and the wavelength coverage of its fluorescence is 640nm ~ 850nm.
In order to reduce cost and to reduce size, make system can be applicable to on-the-spot the detection, the present invention adopts LED as light source 1.The fluorescence excitation spectrum of phycocyanin is near 620nm.
The length of described oval hot spot 14 minor axises is that the length of 20 ± 5um, major axis is 200 ± 20um; The direction of oval hot spot minor axis is consistent with the cell flow direction, and oval hot spot long axis direction is perpendicular to cell flow direction and direction of beam propagation.As shown in Figure 7.The size requirements of oval hot spot 14 in the flow chamber center is: less than 2 blue-green algae diameters, be not less than hole width in the flow chamber on the direction that flows perpendicular to beam propagation and sample at the same time on the sample flow direction.On perpendicular to the direction of beam propagation, light signal is collected on the photodetector 8 with second condenser lens 7 of a large-numerical aperture,
Described photodetector 8 adopts photoelectric diode.In order to eliminate bias light, obtain good signal-to-noise, before photodetector 8, increase a long logical color filter 10, bias light (mainly being irradiates light) is suppressed to greatest extent, allow fluorescence pass through simultaneously.At the excitation spectrum and the characteristic spectral emission of phycocyanin, the curve of spectrum of long logical color filter 10 as shown in Figure 8: less than 0.01%, the percent of pass of 640nm ~ 850nm wavelength is greater than 80% less than the wavelength percent of pass of 620nm.
4, signal Processing and data analysis part: the electric signal of signal Processing and 200 pairs of photodetectors of data analysis unit carries out Filtering Processing, and the statistics of sample analyzed: data analysing method is divided at least 2 zones in the blue-green algae statistics of one dimension at least, the different physiology phases of representing blue-green algae respectively, blue-green algae cell in the zone is counted respectively and analyzed, obtain the blue-green algae concentration of the blue-green algae total concentration of sample liquid, different physiology phases, the shared number percent of different physiology phase blue-green algaes; Described blue-green algae total concentration is the set of all single blue-green algae cell measurement numbers in the tested sample liquid.So just realized the detection of individual cells in the sample.Detect up to ten thousand blue-green algae cells, forming statistical property is exactly the general characteristic of this sample.
In the present invention, not only can carry out quantitative measurment, further, can carry out analysis and judgement the different physiology phase of blue-green algae to the concentration of blue-green algae in the waters.Data analysis module is added up the fluorescence volume of single blue-green algae cell together, forms histogram, as shown in Figure 9, divides three zones---growth period, luxuriant phase, maturity stage on histogram.
Fig. 9 has provided the embodiment that only analyzes with the one dimension fluorescence signal.
For frustule, content growth period of phycocyanin〉the luxuriant phase maturity stage, therefore on fluorescence volume the maturity stage＜luxuriant phase＜growth period.The excitation wavelength peak value 620nm of phycocyanin, emission wavelength peak 640nm.
The transverse axis of Fig. 9 is the fluorescence volume fl of particle, and the longitudinal axis is that fluorescence volume is the number n of the particle of fl.The fluorescence that produces when a blue-green algae cell enters area of illumination along with sample flow is received by photodetector 8, and the final signal that forms in signal Processing and data analysis unit 200 is fl
i, then in histogram shown in Figure 9, fluorescence volume is fl
iThe longitudinal axis height n in place
iIncrease by 1.By that analogy, each blue-green algae cell all can increase by 1 at the longitudinal axis height (number) that its corresponding transverse axis (fluorescence volume) is located by detection zone the time, has just formed histogram shown in Figure 9 when finishing when the blue-green algae cell of all tested samples all detects.
Signal Processing is provided with different threshold value th with the fluorescence volume direction of data analysis unit 200 in histogram
Bk, th
Ma, th
Gr, when being in, the fluorescence volume of blue-green algae cell just reflected this physiological status of cells in certain threshold value, be in th as the fluorescence volume of certain particle
Bk, and th
MaBetween think that then it is in maturity stage M, if the fluorescence volume of cell is positioned at th
MaAnd th
GrBetween think that then it is in maturity stage B, if the fluorescence volume of cell is greater than th
GrThink that then it is in G in growth period.M, B, G just represent different physiological periods---the maturity stage of blue-green algae, luxuriant phase and growth period.
Signal Processing and data analysis unit 200 are in the same place the optical quantities statistics of single blue-green algae cell, form the multidimensional data space, on data space with the several zones of specific boundary demarcation---maturity stage M, luxuriant phase B, growth period G.Drop on three blue-green algae cell numbers in the zone and be respectively N
M, N
B, N
G, the volume of supposing tested sample be V then total blue-green algae concentration of this tested sample be:
C=(N
M+N
B+N
G)/V
The blue-green algae concentration of different physiology phases is respectively:
C
M=N
M/V
C
B=N
B/V
C
G=N
G/V
The number percent that the blue-green algae of different physiology phases accounts for this waters blue-green algae total amount is:
%M=N
M/(N
M+N
B+N
G)
%B=N
B/(N
M+N
B+N
G)
%G=N
G/(N
M+N
B+N
G)。
In Fig. 9, the histogram entire area is represented all blue-green algae number of cells in this sample, can obtain the blue-green algae total concentration of sample to be tested divided by sample volume.With the different physiology phase of different threshold value sign blue-green algaes, the area under each zone is exactly the blue-green algae total cellular score in the Ben Shengli phase.Use C, C
M, C
B, C
G, %M, %B, %G characterize the blue-green algae situation in this sheet waters.
Utilize above parameter to judge to concentration and each physiology phase concentration of this waters blue-green algae.When the maturity stage, the blue-green algae cell concentration was too high, C
MVery big, indicating that blue-green alga bloom is about to outburst, the blue-green algae content that just means the one's respective area when the blue-green algae excessive concentration in luxuriant phase and growth period has the trend of increase, should take measures to suppress, and general inhibition method enters for nutritive salt such as restriction nitrogen, phosphorus.Therefore not only can measure by analytic system of the present invention, and can analyze the upgrowth situation of blue-green algae in this waters to blue-green algae concentration.
Another analytical approach is to divide two zones on data space---maturity stage M and growth period G, computing method are the same.By data presentation and on data the zoning, in zones of different, count, add up, realize that to the analysis of physiology phase of sample blue-green algae can obtain than only the total counting number more information of blue-green algae, the blue-green algae that helps comprehensively to grasp the waters is dynamic.
Use the present invention in once complete measuring process, each " incident " only detects a cell, obtains the character of whole sample by the statistical property of a plurality of incidents, makes measurement more accurate, need not calibrate.And not only obtain blue-green algae total concentration in the sample, and can also analyze blue-green algae concentration and the ratio of different physiology phase, grasp more sample information, help the growing state of the blue-green algae in this waters is carried out early warning.
Claims (9)
1. the check and analysis method of a blue-green algae is characterized in that: comprise following check and analysis step:
(1), sheath stream forms: measurand is formed by two kinds of liquid, and a kind of is sample to be tested liquid, at first gathers water sample with sample-sucking device, and particle diameter is greater than the sand grain of 1mm in the filtering water sample; Inner through screening deaeration and sand grains at sample-sucking device then, remaining phytoplankton is as sample to be tested liquid; Another kind is a sheath fluid; By inhaling sample and liquid road control module two kinds of liquid are delivered in the flow chamber, sample to be tested liquid is at the dirty via flow of the parcel of sheath fluid chamber detection zone, under the effect of fluid focus, sample liquid is compressed to the width of 2 blue-green algae cells, makes the blue-green algae cell one by one by the flow chamber detection zone;
(2), rayed part:
Light source forms an oval hot spot through first condenser lens or lens combination, and oval hot spot shines on the sample to be tested liquid that flows through in the flow chamber detection zone; Sheath fluid forms laminar flow in the flow chamber commutating zone, in the flow chamber accelerating region, sample liquid is focused on gradually, be sample flow until in the flow chamber detection zone, shortening sample to be tested hydraulic pressure into diameter less than 2 blue-green algae cell yardsticks, blue-green algae cell in the sample to be tested liquid just can only pass through the irradiation of detection zone irradiates light one by one like this, can only detect a cell in each detection incident;
(3), light-receiving probe portion: the blue-green algae cell one by one is subjected to the irradiation of detection zone irradiates light and produces optical signalling, and described optical signalling is collected by second condenser lens, enters photodetector by long logical color filter again; Optical signalling is received by photodetector, form electric impulse signal through after the opto-electronic conversion, deliver to signal Processing and data analysis unit, a stream of cells just forms an electric impulse signal when crossing detection zone, has so just realized the measurement one by one of individual cells in the sample;
(4) signal Processing and data analysis part: signal Processing and data analysis unit are carried out Filtering Processing to the electric signal of photodetector, and the statistics of sample analyzed, in the blue-green algae statistics of one dimension at least, divide at least 2 zones according to analytical approach, the different physiology phases of representing blue-green algae respectively, blue-green algae cell in the zone is counted respectively and analyzed, obtain the blue-green algae concentration of the blue-green algae total concentration of sample liquid, different physiology phases, the shared number percent of different physiology phase blue-green algaes; The total number that described blue-green algae total concentration is all pulses in the one-shot measurement process is divided by tested volume.
2. the check and analysis method of a kind of blue-green algae as claimed in claim 1, it is characterized in that: described flow chamber comprises the guide hole that a flow of liquid is crossed, and this guide hole is divided into three zones: commutating zone, accelerating region, detection zone; Described detection zone is that the rayed zone is square hole or the rectangular opening that a length of side is 200um～400um, is made by optically transparent material; The part of the inlet flaring of described square hole or rectangular opening forms accelerating region, and the part that sheath fluid passage and sample liquid passage are nested is a commutating zone.
3. the check and analysis method of a kind of blue-green algae as claimed in claim 2, it is characterized in that: described sample liquid channel outlet is the circular hole of diameter 0.3mm.
4. the check and analysis method of a kind of blue-green algae as claimed in claim 1 is characterized in that: described light source employing light emitting diode; Described photodetector adopts photodiode.
5. the check and analysis method of a kind of blue-green algae as claimed in claim 4, it is characterized in that: the wavelength 620 ± 5nm of described light emitting diode, printing opacity wavelength are 640nm～850nm.
6. the check and analysis method of a kind of blue-green algae as claimed in claim 1, it is characterized in that: the liquid stream of described flow chamber internal flow will satisfy laminar flow condition, i.e. Reynolds number＜2300; Sheath stream carries out fluid focus to sample flow, and sample flow is compressed to width less than 2 blue-green algae cell dias.
7. the check and analysis method of a kind of blue-green algae as claimed in claim 1, it is characterized in that: the length of described oval hot spot minor axis is that the length of 20 ± 5um, major axis is 200 ± 20um; The direction of oval hot spot minor axis is consistent with the cell flow direction, and oval hot spot long axis direction is perpendicular to cell flow direction and direction of beam propagation.
8. the check and analysis method of a kind of blue-green algae as claimed in claim 1, it is characterized in that: described oval hot spot in the size requirements of flow chamber center is: less than 2 blue-green algae diameters, be not less than hole width in the flow chamber on the direction that flows perpendicular to beam propagation and sample at the same time on the sample flow direction; On perpendicular to the direction of beam propagation, light signal is collected on the photodetector with second condenser lens of a large-numerical aperture.
9. the check and analysis method of a kind of blue-green algae as claimed in claim 1, it is characterized in that: the optical signalling that produces after described blue-green algae cell is subjected to shining comprises: forward scattering light, side scattered light and fluorescence, described forward scattering light refers to the scattered light in 1 ~ 5 degree scope along the light path direction of propagation, has reflected the volume size of cell; Side scattered light refers to the scattered light perpendicular to optical propagation direction, and angular range is 75 degree ~ 115 degree, has reflected the inside complexity of cell; The angular range of fluorescence is consistent with the side scattered light angle, and the wavelength coverage of its fluorescence is 640nm ~ 850nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010598251 CN102095686A (en) | 2010-12-21 | 2010-12-21 | Method for detecting and analyzing blue-green algae |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010598251 CN102095686A (en) | 2010-12-21 | 2010-12-21 | Method for detecting and analyzing blue-green algae |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102095686A true CN102095686A (en) | 2011-06-15 |
Family
ID=44128867
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010598251 Pending CN102095686A (en) | 2010-12-21 | 2010-12-21 | Method for detecting and analyzing blue-green algae |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102095686A (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104713839A (en) * | 2013-12-13 | 2015-06-17 | 中国科学院大连化学物理研究所 | Automatic online microalgae OD determinator |
| CN105527268A (en) * | 2016-02-26 | 2016-04-27 | 中国科学院南京地理与湖泊研究所 | Surface sediment phycocyanin in-situ fast detecting device and system |
| CN105527260A (en) * | 2015-12-21 | 2016-04-27 | 江南大学 | Online detection device of concentration of blue-green algae in water body |
| CN106053302A (en) * | 2016-07-25 | 2016-10-26 | 中国科学院苏州生物医学工程技术研究所 | System for detecting floating algae |
| CN107192657A (en) * | 2017-06-29 | 2017-09-22 | 太湖流域水文水资源监测中心（太湖流域水环境监测中心） | A kind of algae on-line detecting system |
| CN108318424A (en) * | 2018-04-09 | 2018-07-24 | 浙江大学 | A kind of underwater planktonic organism automatic imaging device and imaging method |
| CN110617804A (en) * | 2019-09-25 | 2019-12-27 | 浙江海洋大学 | Marine ecological environment detection system and method based on remote sensing technology |
| CN113337385A (en) * | 2021-08-05 | 2021-09-03 | 于吉(南京)生物科技有限公司 | Automatic monitoring method and device for cell culture |
| CN115015178A (en) * | 2022-08-05 | 2022-09-06 | 天津迈科隆生物科技有限公司 | Optical detection device and blood analyzer |
| CN116076348A (en) * | 2014-03-04 | 2023-05-09 | 绿玛瑙有限公司 | System and method for growing and distributing aquatic organisms |
| CN118421457A (en) * | 2024-04-29 | 2024-08-02 | 广东海洋大学 | A microalgae density detection system and method based on visible light communication |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101153868A (en) * | 2006-09-30 | 2008-04-02 | 深圳迈瑞生物医疗电子股份有限公司 | Stream type cell analyzer |
| CN101275898A (en) * | 2007-03-29 | 2008-10-01 | 希森美康株式会社 | Biological sample analyzer |
| CN101285824A (en) * | 2008-04-22 | 2008-10-15 | 济南市供排水监测中心 | Blue algae polluted water microcystic toxins checking method |
| CN101726454A (en) * | 2008-10-28 | 2010-06-09 | 深圳市普康电子有限公司 | Sheath flow chamber of blood cell analyzer |
| US20100248293A1 (en) * | 2009-03-31 | 2010-09-30 | Sysmex Corporation | Sample analyzer and sample analyzing method |
-
2010
- 2010-12-21 CN CN 201010598251 patent/CN102095686A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101153868A (en) * | 2006-09-30 | 2008-04-02 | 深圳迈瑞生物医疗电子股份有限公司 | Stream type cell analyzer |
| CN101275898A (en) * | 2007-03-29 | 2008-10-01 | 希森美康株式会社 | Biological sample analyzer |
| CN101285824A (en) * | 2008-04-22 | 2008-10-15 | 济南市供排水监测中心 | Blue algae polluted water microcystic toxins checking method |
| CN101726454A (en) * | 2008-10-28 | 2010-06-09 | 深圳市普康电子有限公司 | Sheath flow chamber of blood cell analyzer |
| US20100248293A1 (en) * | 2009-03-31 | 2010-09-30 | Sysmex Corporation | Sample analyzer and sample analyzing method |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104713839A (en) * | 2013-12-13 | 2015-06-17 | 中国科学院大连化学物理研究所 | Automatic online microalgae OD determinator |
| CN104713839B (en) * | 2013-12-13 | 2017-04-05 | 中国科学院大连化学物理研究所 | A kind of microalgae automatic on-line OD analyzers |
| CN116076348A (en) * | 2014-03-04 | 2023-05-09 | 绿玛瑙有限公司 | System and method for growing and distributing aquatic organisms |
| CN105527260A (en) * | 2015-12-21 | 2016-04-27 | 江南大学 | Online detection device of concentration of blue-green algae in water body |
| CN105527268A (en) * | 2016-02-26 | 2016-04-27 | 中国科学院南京地理与湖泊研究所 | Surface sediment phycocyanin in-situ fast detecting device and system |
| CN106053302A (en) * | 2016-07-25 | 2016-10-26 | 中国科学院苏州生物医学工程技术研究所 | System for detecting floating algae |
| CN107192657A (en) * | 2017-06-29 | 2017-09-22 | 太湖流域水文水资源监测中心（太湖流域水环境监测中心） | A kind of algae on-line detecting system |
| CN108318424A (en) * | 2018-04-09 | 2018-07-24 | 浙江大学 | A kind of underwater planktonic organism automatic imaging device and imaging method |
| CN110617804A (en) * | 2019-09-25 | 2019-12-27 | 浙江海洋大学 | Marine ecological environment detection system and method based on remote sensing technology |
| CN113337385A (en) * | 2021-08-05 | 2021-09-03 | 于吉(南京)生物科技有限公司 | Automatic monitoring method and device for cell culture |
| CN115015178A (en) * | 2022-08-05 | 2022-09-06 | 天津迈科隆生物科技有限公司 | Optical detection device and blood analyzer |
| CN118421457A (en) * | 2024-04-29 | 2024-08-02 | 广东海洋大学 | A microalgae density detection system and method based on visible light communication |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102095686A (en) | Method for detecting and analyzing blue-green algae | |
| CN101498646B (en) | Forward-scattering signal inspection device and method, cell or particle analyzer | |
| CN111610175B (en) | Flow-through type phytoplankton type and cell density detection device and detection method | |
| CN102279174B (en) | Alga identification and measurement sensor and method | |
| CN102192872B (en) | Optical measuring device and optical measuring method | |
| JP6019039B2 (en) | Electro-optic flow measuring device | |
| KR101264075B1 (en) | Optical Apparatus for Measuring Particles | |
| JP5308521B2 (en) | Electro-optical measuring apparatus and electro-optical measuring method for classifying and counting fine components | |
| CN102331411A (en) | Blood cell analyzer with blue semiconductor laser | |
| US20110310386A1 (en) | Method and system for analysing solid particles in a medium | |
| EP2694668A1 (en) | Microbial detection apparatus and method | |
| CN202177564U (en) | Blood cell analyzer adopting blue semiconductor laser | |
| CN104807738A (en) | Device for detecting shapes of single aerosol particles in real time | |
| EP3701235B1 (en) | A fluorescent substance detection system | |
| CN112485167A (en) | Optical system of particle analyzer | |
| CN107831099A (en) | The detection means and detection method of air particle | |
| CN112285035B (en) | A water color recognition device and a water color recognition method | |
| CN102331397A (en) | Photoelectric sensor for statistic analysis of blood cells | |
| CN108956402A (en) | A kind of highly sensitive dust concentration detecting method with compound how photosensitive plot structure | |
| CN112903547A (en) | High-concentration cloud and mist particle concentration measuring device based on double light sources | |
| CN105628658A (en) | Optical detection system for bioaerosol and detection method | |
| CN102095712A (en) | Photoelectric sensor for detecting blue-green algae | |
| CN206557053U (en) | A kind of oil quality detection means | |
| CN117825220A (en) | Sensitization aerosol monitor and method | |
| JP2000241335A (en) | Algae and microparticle counting method and counting device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| AD01 | Patent right deemed abandoned |
Effective date of abandoning: 20110615 |
|
| C20 | Patent right or utility model deemed to be abandoned or is abandoned |