CN107314966A - A kind of flow cytometer algae on-line detecting system and method - Google Patents
A kind of flow cytometer algae on-line detecting system and method Download PDFInfo
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- CN107314966A CN107314966A CN201710513141.7A CN201710513141A CN107314966A CN 107314966 A CN107314966 A CN 107314966A CN 201710513141 A CN201710513141 A CN 201710513141A CN 107314966 A CN107314966 A CN 107314966A
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- 241000195493 Cryptophyta Species 0.000 title claims abstract description 101
- 238000000034 method Methods 0.000 title claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 246
- 238000001514 detection method Methods 0.000 claims abstract description 30
- 238000003760 magnetic stirring Methods 0.000 claims abstract description 29
- 238000005070 sampling Methods 0.000 claims abstract description 19
- 238000010998 test method Methods 0.000 claims abstract description 13
- 230000002572 peristaltic effect Effects 0.000 claims description 17
- 238000002347 injection Methods 0.000 claims description 13
- 239000007924 injection Substances 0.000 claims description 13
- 230000004087 circulation Effects 0.000 claims description 11
- 239000012530 fluid Substances 0.000 claims description 9
- 238000002203 pretreatment Methods 0.000 claims description 8
- 239000000049 pigment Substances 0.000 claims description 6
- 238000000605 extraction Methods 0.000 claims description 5
- 238000007667 floating Methods 0.000 claims description 5
- 241000009298 Trigla lyra Species 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 3
- 238000002604 ultrasonography Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 4
- 238000011065 in-situ storage Methods 0.000 abstract description 3
- 238000012544 monitoring process Methods 0.000 description 11
- 238000007689 inspection Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 239000005416 organic matter Substances 0.000 description 5
- 230000005611 electricity Effects 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000178 monomer Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 238000005086 pumping Methods 0.000 description 4
- 241000192700 Cyanobacteria Species 0.000 description 3
- 241000192710 Microcystis aeruginosa Species 0.000 description 3
- 238000013019 agitation Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000009183 running Effects 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000005446 dissolved organic matter Substances 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 238000009413 insulation Methods 0.000 description 2
- 238000013507 mapping Methods 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000037452 priming Effects 0.000 description 1
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- 239000002699 waste material Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1425—Optical investigation techniques, e.g. flow cytometry using an analyser being characterised by its control arrangement
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Abstract
The invention discloses a kind of algae on-line detecting system and method, the system includes water inlet pipe, one end of the water inlet pipe is stretched into collection water sample, the other end of the water inlet pipe is connected with pond, it is connected with magnetic valve and the water pump for sampling on water inlet pipe between collection water sample and the pond in turn, the bottom in the pond is provided with magnetic stirring apparatus, the side in the pond is provided with ultrasonic cell disintegration instrument, the ultrasonic cell disintegration instrument is connected by cable with one end of ultrasonic transformer, the other end of the ultrasonic transformer is stretched into the pond, level sensor is provided with the pond, the pond is connected by sample feeding pipe with the CytoSense flow cytometers detected for algae, the sidewall bottom in the pond is connected with drainpipe, the side wall upper part in the pond is connected with overflow pipe；The invention discloses a kind of algae online test method, the system and method have the advantages that detection efficiency it is high, can real-time in-situ detection, testing result it is accurate, applied widely.
Description
Technical field
The invention belongs to algae detection technique field, and in particular to a kind of flow cytometer algae on-line detecting system and side
Method.
Background technology
Want to carry out accurate early warning before breakout of cyanobacteria blooms, it is conscientiously may be used to continuously monitor blue-green algae content in water
Capable approach.Current continuous monitoring relies primarily on flow cytometer to monitor analysis, the general principle of flow cytomery
For：The particle (including cell) allowed according to principle of hydrodynamics in liquid passes through laser beam (detection zone), laser irradiation one by one
The scattering of light can be caused on to particle, such as fruit granule (such as frustule) can be sent out fluorescence containing pigment, these scattered lights and
Fluorescence is converted into electric signal after being collected by detector and stored, and is automatically analyzed using software.Phytoplankton in water
Cell is used as particle one by one, it is of course possible to carry out flow cytometry, thus can monitor the content of blue-green algae in water.
Patent application publication number discloses for CN102128779A and CN102128776A Chinese invention patent application
A kind of method of use Flow cytometry algae and chromophoric dissolved organic matter, General Matters are as follows：Using injector
Water sample is gathered, through filter cleaner, then algae to be detected and organic matter are detected using flow cytometry, and will detection
Parameter number in algae or organic matter cell size and unit volume is tried to achieve by certain algorithm.Patent application is public
Cloth number for CN102128779A Chinese invention patent application carry out Flow cytometry when, disclose algae to be detected and
Chromophoric dissolved organic matter is wrapped in by the non-external sheath fluid without any particle flows through flow chamber, is expected that by non-external sheath fluid
Hydrodynamics focussing force make the algae to be detected and the organic matter single-row layout under the coating of sheath fluid, pass sequentially through algae
Class detection zone, during this, intergranular distance is opened, and it is unicellular that detection is only radiated at an algae with laser every time
Or on an organic matter cell, to reach the effect for improving detection accuracy.But, due to not entering to the liquid sample of collection
The scattered pretreatment of row, is only extremely difficult to individual cells by way of non-external sheath fluid parcel and separates and pass sequentially through laser detection
The effect in area, in addition, above-mentioned two application for a patent for invention does not disclose the concrete mode of sampling, does not disclose invention described yet
The applicable waters scope of sample mode, it detects that algae especially algae big to aggregate density not necessarily has universality, together
When, above-mentioned sample mode is possible to kill alga cells in operation, can cause testing result accuracy further under
Drop.
In addition, patent application publication number discloses a kind of streaming in detail for CN106053302A Chinese invention patent application
Cell art detection device, but the pre-treating method and specific sampling process of collecting sample are not disclosed.
The conventional algae detection method in the country is to gather algae sample using manual site, and laboratory utilization is taken back after fixing
Microscope imaging technology is observed.Micro imaging method includes optical microscope, electron microscopic observation, fluorescence microscopy method etc..
Optical microscope is distinguishable to kind, but pre-treatment work is complicated, and sample identification must be entered by veteran taxology expert
OK, time and effort consuming；Electronic Speculum multiplication factor is high, is conducive to distinguishing doubtful kind, but sample preparation and operation qualification process are excessively complicated；
Fluorescence microscopy observes the biology for being then used for autofluorescence.At present, the domestic algae monitoring carried out also is in the conventional way
It is main, because taxonomic identification workload is larger, it is difficult to adapt to the requirement that field batch samples are quick and precisely analyzed, and can not carry out
Real-time monitoring in situ, in actual applications with certain limitation.
The content of the invention
The present invention provides a kind of flow cytometer algae on-line detecting system and method for the defect of prior art.
The purpose of the present invention is achieved through the following technical solutions：
A kind of algae on-line detecting system, including water inlet pipe, one end of the water inlet pipe are stretched into collection water sample, it is described enter
The other end of water pipe is connected with pond, positioned at collection water sample and the pond between water inlet pipe on be connected with turn magnetic valve and
For the water pump of sampling, the bottom in the pond is provided with magnetic stirring apparatus, and the side in the pond is provided with ultrasonic cell and broken
Broken instrument, the ultrasonic cell disintegration instrument is connected by cable with one end of ultrasonic transformer, and the other end of the ultrasonic transformer stretches into described
In pond, level sensor is provided with the pond, the pond is flowed by water pipe with the CytoSense for algae detection
Formula cell instrument is connected, and the sidewall bottom in the pond is connected with drainpipe, and the side wall upper part in the pond is connected with overflow pipe；
The magnetic stirring apparatus, ultrasonic cell disintegration instrument, CytoSense flow cytometers, level sensor, magnetic valve, water pump difference
Electrically connected with electronic control box, the electronic control box is electrically connected with terminal computer.
A kind of online test method of algae on-line detecting system according to claim 1, comprises the following steps：
1) sample：The control software of terminal computer is opened, control opens the magnetic valve and water pump, passes through the water pump control
Water source sample in water inlet pipe collection sampling water source processed, pumps quantitative water source sample into the pond at regular intervals,
Acquisition rate is 0.5-2L/min；
2) pre-treatment：When the water source sample reaches predetermined amount, by opening the control software of terminal computer, set
The magnetic stirring apparatus, the relevant parameter of ultrasonic cell disintegration instrument and operation, the terminal computer are sent out to the electronic control box
Electric signals, the electronic control box controls magnetic stirring apparatus, ultrasonic cell disintegration instrument to be operated according to default parameter respectively, institute
State the water source sample that magnetic stirring apparatus is pointed in pond to be stirred, it is ensured that the uniformity at sample water source, avoid algae from floating；
The ultrasonic cell disintegration instrument carries out ultrasonic vibration by coupled ultrasonic transformer to the water source sample in pond, reaches fully
The purpose of scattered algae；
3) detect：When the pretreatment process is completed, by the control software of terminal computer, control is described
CytoSense flow cytometers bring into operation, and the CytoSense flow cytometers can be by regular time and quantity out of described pond
Extract the sample water source by pretreatment, and the sample water source of extraction is completed the number of algae, alga cells length, size,
Form, granularity, pigment, peak number, algae group character detect and draw corresponding pulse collection of illustrative plates.
Further, the water pump is peristaltic pump；The water inlet pipe stretch into sample water source bifurcated at one end formed it is multiple enter
Water branch pipe, the magnetic valve has multiple, and multiple water inlet pipes are connected by a magnetic valve with the water inlet pipe respectively；
Multiple water inlet pipes are respectively protruding at the different depth of sample water source vertical direction.
Further, the water inlet pipe stretches into first water inlet pipe of bifurcated at one end formation at sample water source, the second water inlet branch
Pipe and the 3rd water inlet pipe, the magnetic valve include the first magnetic valve, the second magnetic valve, the 3rd magnetic valve and the 4th magnetic valve,
First water inlet pipe, the second water inlet pipe and the 3rd water inlet pipe pass through the first magnetic valve, the second magnetic valve, the 3rd respectively
Magnetic valve is connected with water inlet pipe, and the water inlet pipe is connected with the 4th magnetic valve and silt positioned at one section between peristaltic pump and pond
Filter；First water inlet pipe is located at sample water source bottom 0.5m, and second water inlet pipe is located at sample water source bottom
1m, the 3rd water inlet pipe is located at sample water source bottom 1.5m.
Further, in sampling step, first magnetic valve, the second magnetic valve, the 3rd electromagnetism are selectively turned on
One or several and the 4th magnetic valve in valve, can realize the inspection of algae at the different depth to sample water source vertical direction
Survey.
Further, in pre-treatment step, pretreatment is started when the pond water is 1-3L；Control the magnetic force
The revolution of agitator is that 1000-3000 turns/min；The power for controlling the ultrasonic cell disintegration instrument is 300-660W, the ultrasound
Cell crushing instrument handles 10-33 circulation, and each circulation includes ultrasonic vibration 3-10s, stops 3-15s.
Further, in detecting step, control the CytoSense flow cytometers enters pump rate for 0.5-2 μ l/
S, collecting sample water source amount is 0.1-0.5ml, and the CytoSense flow cytometer sheath fluids choose pure water.
Further, the algae on-line detecting system also includes sound proof box, the ultrasonic transformer, magnetic stirring apparatus and pond
In the sound proof box.
Further, the side wall upper part in the pond is also associated with hot water injection's pipe, is also associated with the water inlet pipe cold
Water flush pipe, is connected with communicating pipe between hot water injection pipe and the cold water flush pipe, hot water injection's pipe, described cold
Water flush pipe is connected to the 5th magnetic valve and the 6th magnetic valve at the pipe end positioned at the communicating pipe side, described
Hot water injection's pipe, the cold water flush pipe have been also respectively connected with control piper switch on the pipeline positioned at the communicating pipe side
The 7th magnetic valve and the 8th magnetic valve.
Further, the bottom of the flow cytometer is provided with exhausted bath box.
Compared with prior art, the present invention at least has advantages below：
1. algae on-line detecting system of the present invention replaces simple self priming pump in sampling process using peristaltic pump, to the greatest extent
The destruction to algae in sampling process is possibly reduced, monitoring result is more accurate.And traditional algae monitoring technology then needs profit
Algae is fixed with fixative, frustule has been damaged, and jolting during transport and preservation also can be to alga cells
Impact；
2nd, the present invention innovatively adds water sample pre-treatment, is broken up the big group of algae for groupuscule very by physical method
To monomer, be conducive to improving the count accuracy of flow cytometer, sampled water be stirred by magnetic stirring apparatus, it is to avoid
Algae, which floats, influences the accuracy of detection, meanwhile, magnetic agitation is destroyed with scattered will not be produced to algae of ultrasonic vibration；
3rd, the present invention can gather collection algae sample at water source vertical direction different water depth, to obtain water body vertical direction
The feature and changing rule of amount of algae, population structure at upper diverse location；
4th, the water sample of different algae density is detected that the system is working properly in present invention selection Taihu Lake, shows the algae
On-line monitoring system is suitable for the water body of high concentration algae；The flow cytometer algae on-line monitoring system is applied to national model
The algae on-line monitoring of freshwater lake/reservoir in enclosing, it is applied widely；
5th, flow cytometer algae on-line detecting system of the invention is combined with tele-control system, facilitates policymaker's root
Judge according to the detection for needing to make number and population structure of the algae of monitored area etc. real-time in-situ, and to algae change
Trend is made a prediction, and can either be made the decision of detection by terminal computer and be set relevant parameter, realizes algae detection oneself
Dynamicization is operated, convenient and swift；
6th, flow cytometer algae on-line detecting system of the invention is more efficient compared with traditional algae monitoring technology,
This means algae on-line detecting system can be gathered and analyze and process more algae samples；
7th, flow cytometer algae on-line detecting system of the invention uses the pattern of full instrumentation, with traditional algae
Monitoring technology needs the situation of artificial micro- sem observation alga cells number and population structure to compare, testing result by it is subjective because
The probability of element interference is relatively low, and accuracy and reliability are higher；
8th, the present invention compared with the flow cytometer of other models, CytoSense flow cytometers it is simple to operate, be suitable to
The algae scope of detection is wide, while the device of the present invention applies also for the detection of organic matter etc., it is applied widely.
Brief description of the drawings
Fig. 1 is the schematic diagram of the mechanical structure of algae on-line detecting system of the present invention；
Fig. 2 is the signal connection diagram of algae on-line detecting system of the present invention.
Wherein, 1- algae on-line detecting system；2- water inlet pipes；The water inlet pipes of 3- first；The water inlet pipes of 4- second；5- the 3rd
Water inlet pipe；The magnetic valves of 6- first；The magnetic valves of 7- second；The magnetic valves of 8- the 3rd；9- peristaltic pumps；The magnetic valves of 10- the 4th；11- water
Pond；12- magnetic stirring apparatus；13- ultrasonic cell disintegration instrument；14- cables；15- ultrasonic transformers；16- sample feeding pipes；17-CytoSense flows
Formula cell instrument；18- overflow pipes；19- drainpipes；20- columns；21- stations；22- sound proof boxs；23- silt filters；24- cold water
Flush pipe；25- hot water injections manage；The magnetic valves of 26- the 5th；The magnetic valves of 27- the 6th；The magnetic valves of 28- the 7th；The magnetic valves of 29- the 8th；
30- communicating pipes；31- exhausted bath box；32-CytoUSB inspection softwares；33- on-line checking softwares；34- electronic control box；35- terminals
Computer；36-Cytoclus mapping softwares.
Embodiment
The invention will be further described with reference to the accompanying drawings and examples, and following examples are descriptive, are not
Limited, it is impossible to protection scope of the present invention is limited with this.
Embodiment 1
As shown in figure 1, a kind of algae on-line detecting system 1, the algae on-line detecting system 1 is located at the station at sample water source
In room 21, the station 21 is supported on sample water source by column 20, and the algae on-line detecting system 1 includes water inlet pipe
2, one end of the water inlet pipe 2 is stretched into collection water sample, and the bifurcated at one end formation first that the water inlet pipe 2 stretches into sample water source is entered
Water branch pipe 3, the second water inlet pipe 4 and the 3rd water inlet pipe 5, first water inlet pipe 3 are located at sample water source bottom 0.5m, institute
The second water inlet pipe 4 is stated positioned at sample water source bottom 1m, the 3rd water inlet pipe 5 is located at sample water source bottom 1.5m；Also wrap
The first magnetic valve 6, the second magnetic valve 7, the 3rd magnetic valve 8 are included, first water inlet pipe 3, the second water inlet pipe 4 and the 3rd enter
Water branch pipe 5 is connected by the first magnetic valve 6, the second magnetic valve 7, the 3rd magnetic valve 8 with the water inlet pipe 2 respectively, and above-mentioned first
Water inlet pipe 3, the second water inlet pipe 4 and the 3rd water inlet pipe 5 are respectively used to the water source of different depth under sample drawn water source；Institute
The other end for stating water inlet pipe 2 is connected with pond 11, is connected with turn on the water inlet pipe 2 being connected with the pond 11 for sampling
Peristaltic pump 9, the 4th magnetic valve 10 and silt filter 23, the peristaltic pump 9 are taken out by way of compacted suction into the pond 11
This water source is sampled, the destruction to algae in collection water sample can be avoided, the silt filter is used to filter in collection water sample
Silt；In the present embodiment, the first magnetic valve 6 and the 4th magnetic valve being connected with the first water inlet pipe 3 are selectively turned on
10, realize the detection to sample water source vertical direction bottom 0.5m depth algae.
The bottom in the pond 11 is provided with magnetic stirring apparatus 12, and the magnetic stirring apparatus is used for pond described in magnetic agitation
Collection water sample in 11, the side of pond 11 is provided with ultrasonic cell disintegration instrument 13, and the ultrasonic cell disintegration instrument 13 passes through
Cable 14 is connected with one end of ultrasonic transformer 15, and the other end of the ultrasonic transformer 15 is stretched into the pond 11, and the ultrasonic transformer is used
Collection water sample inside to the pond carries out ultrasonic agitation, and the pond 11 is detected by sample feeding pipe 16 with being used for algae
CytoSense flow cytometers 17 are connected, the sample feeding pipe 16 be used for the pond 11 inside extract by stirring and
Ultrasonically treated collection water sample, the sidewall bottom in the pond 11 is connected with drainpipe 19, and the drainpipe 19 is used for after testing
The water sample inside the pond 11 is excluded, the side wall upper part in the pond 11 is connected with the overflow pipe 18 for overflow, described to overflow
Flow tube 18 is used to exclude in the water of overflow inside the pond, the level sensor provided with monitoring water level inside the pond, when
When water level in the pond reaches requirement, the level sensor will be to the electronic equipment (Electronic Control i.e. hereafter
Case 34) signal is sent, the first magnetic valve, the 4th magnetic valve and peristaltic pump are closed by the electronic equipment, stopped pumping；
The magnetic stirring apparatus 12, ultrasonic cell disintegration instrument 13, CytoSense flow cytometers 17, level sensor,
One magnetic valve 6, the second magnetic valve 7, the 3rd magnetic valve 8, the 4th magnetic valve 10, peristaltic pump 9, silt filter 23 respectively with electronics
Control cabinet 34 is electrically connected, and the electronic control box 34 is electrically connected with terminal computer 35.
When it is implemented, also include sound proof box 22, the ultrasonic transformer 15, magnetic stirring apparatus 12 and pond 11 be located at it is described every
In audio amplifier 22, the sound proof box 22 plays the effect of sound insulation shock insulation.
A kind of online test method of algae on-line detecting system, comprises the following steps：
1) sample：Selectively turn on the first magnetic valve 6 being connected with the first water inlet pipe 3, the 4th magnetic valve 10 and compacted
Dynamic pump 9, controls the water source sample in water inlet pipe collection sampling water source, at regular intervals to the water by the peristaltic pump 9
Quantitative water source sample is pumped in pond, acquisition rate is 1L/min, the sample water source filters silt by silt filter 23；
2) pre-treatment：When the water source sample reaches 1.5L, stop pumping water source and start pretreatment, first turn on terminal
The control software of computer 35, sets the 1500 turns/min of rotating speed of the magnetic stirring apparatus 12；Control the ultrasonic cell disintegration instrument
13 power is 480W, and the ultrasonic cell disintegration instrument 13 handles 23 circulations, and each circulation includes ultrasonic vibration 3s, stopped
5s；
The water source sample that the magnetic stirring apparatus 12 is pointed in pond 11 is stirred, to ensure the uniform of sample water source
Property, avoid algae in water sample from floating the accuracy of the detection of influence CytoSense flow cytometers 17；The ultrasonic cell disintegration instrument
13 carry out ultrasonic vibration by coupled ultrasonic transformer 15 to the water source sample in pond 11, reach fully dispersed algae especially
It is the purpose of common Microcystis aeruginosa (connected and finally flocked together into bulk by ICC between alga cells) in the body of lake,
The big group of algae is broken up as groupuscule even monomer as far as possible.
3) detect：When the pretreatment process is completed, by the control software of terminal computer 35, set described
CytoSense flow cytometers 17 extract the sample water source for passing through pretreatment by 1 μ l/s of pump speed out of described pond 11, simultaneously
Start detection, the sheath fluid chooses pure water, the sample water source of extraction is completed the number of algae, alga cells length, size,
The parameter detectings such as form, granularity, pigment, peak number, algae colony's feature simultaneously draw corresponding pulse collection of illustrative plates.
The magnetic stirring apparatus 12, ultrasonic cell disintegration instrument 13, CytoSense flow cytometers 17, level sensor,
One magnetic valve 6, the second magnetic valve 7, the 3rd magnetic valve 8, the 4th magnetic valve 10, silt filter 23, peristaltic pump 9 respectively with control
The magnetic stirring apparatus 12, ultrasonic cell disintegration instrument 13, CytoSense flow cytometers 17, level sensor, the first magnetic valve
6th, the Electronic Control that the second magnetic valve 7, the 3rd magnetic valve 8, the 4th magnetic valve 10, silt filter 23, peristaltic pump 9 are operated automatically
Case 34 is electrically connected, and the electronic control box 34 is electrically connected with terminal computer 35, wherein, the level sensor and the electronics control
Two-way electrical connection between case 34 processed, other are unidirectional electrical connection.
As shown in Fig. 2 the control software includes on-line checking software 33, the CytoSense streams that applicant voluntarily researches and develops
CytoUSB inspection softwares 32 and mapping software Cytoclus36 that formula cell instrument 17 is carried, the CytoUSB inspection softwares 32 lead to
Cross terminal computer 35 and CytoSense flow cytometers 17 be two-way electrically connects, the CytoSense flow cytometers 17 with it is described
Electronic control box 34 is unidirectionally electrically connected；The inside of terminal computer 35 is provided with mapping software Cytoclus36, and the drawing is soft
The data that part Cytoclus36 is used to detect the CytoSense flow cytometers 17 are depicted as corresponding chart；It is described
Line inspection software 33 is electrically connected by two-way between terminal computer 35 and the electronic control box 34, the on-line checking software 33
For setting the magnetic stirring apparatus 12, the parameter of ultrasonic cell disintegration instrument 13, the CytoUSB inspection softwares 32 to be used to set
The parameter of CytoSense flow cytometers 17, after relevant parameter is provided with, clicks on operation above-mentioned two software, the end
End computer 35 sends signal to the electronic control box 34, and the electronic control box 34 controls the magnetic stirring apparatus 12, ultrasound
The automatic runnings such as cell crushing instrument 13, CytoSense flow cytometers 17, peristaltic pump 9, magnetic valve.
The annexation and function of the control software of the electronic control box 34 and terminal computer 35 are as follows：
1) the on-line checking software 33 of the electronic control box 34 and the terminal computer 35 transmits mutually data, the electricity
The device data of reception is transferred to on-line checking software 33 by sub- control cabinet 34, and performs the data processing of on-line checking software 33
As a result, the execution is exactly that the data processed result come according to on-line checking software transmission is coupled orderly to run
Equipment.For example, the electronic control box 34 receives the data that level sensor transmits and is transferred to on-line checking software 33, and
The operation of magnetic stirring apparatus 12, ultrasonic cell disintegration instrument 13 is opened according to the result of the feedback of on-line checking software 33.In addition, institute
State electronic control box 34 also had according to the instruction of receiving the first magnetic valve 6 of control, the second magnetic valve 7, the 3rd magnetic valve 8,
The coupled equipment operation such as CytoSense flow cytometers 17, peristaltic pump 9, the 4th magnetic valve 10 or the function of closing, institute
State the electronic control box 34 and control correlation successively according to the step in the online test method of above-mentioned algae on-line detecting system
The operation or closing of equipment.
2) the on-line checking software 33 sets sampling, the operation program of pretreatment in advance, and according in the course of work
The data received from electronic control box 34 to the feedback information of electronic control box 34, the electronic control box 34 according to feedback information come
The operation of the coupled relevant device of control.When sampling, preprocessing process are completed, the on-line checking software 33 is to electronics
Control cabinet 34 sends instruction, and electric signal is sent from electronic control box 34 to CytoSense flow cytometers 17, described
CytoSense flow cytometers 17 transmit the signal for running and starting, the CytoUSB softwares 32 to the CytoUSB softwares 32
Sent to the CytoSense flow cytometers 17 and instruct and control the CytoSense flow cytometers 17 to start according to institute
The detection parameter automatic running that CytoUSB softwares 32 are previously set is stated, to reach the online inspection of above-mentioned algae on-line detecting system
The function of detection in survey method.
3) destination file that the CytoUSB softwares 32 are used to detect the CytoSense flow cytometers 17 is preserved
In specified folder in terminal computer 35, the Cytoclus mapping softwares 36 are opened manually, and paint by the Cytoclus
Figure software 36 transfers the destination file that the CytoSense flow cytometers 17 are detected, and relevant parameter detection data are drawn
Shown into corresponding diagrammatic form.
The side wall upper part in the pond is also associated with being also associated with cold water flush pipe on hot water injection's pipe 25, the water inlet pipe
24, it is connected with communicating pipe 30 between hot water injection's pipe 25 and the cold water flush pipe 24, it is hot water injection's pipe 25, described
Cold water flush pipe 24 is connected to the 5th magnetic valve 26 and the 6th electricity at the pipe end positioned at the side of communicating pipe 30
Magnet valve 27, hot water injection's pipe 25, the cold water flush pipe 24 are also distinguished on the pipeline positioned at the side of communicating pipe 30
It is connected with the 7th magnetic valve 28 and the 8th magnetic valve 29 of control piper switch.When in the water sample algae detection complete with
Afterwards, the 5th magnetic valve 26, the 7th magnetic valve 28, the 8th magnetic valve 29, the first magnetic valve 6, the second magnetic valve 7, are first turned on
Three magnetic valves 8 and the 4th magnetic valve 10, are intake water inlet pipe, pond, silt filter, the first water inlet pipe 3, second with cold water
The alga cells of residual in the water inlet pipe 5 of branch pipe 4 and the 3rd are poured in sampling water source, after complete with cold water flush, are closed
5th magnetic valve 26, opens the 6th magnetic valve 27, with hot water by water inlet pipe, pond, silt filter, the first water inlet pipe, the
Two water inlet pipes and the alga cells of the residual in the 3rd water inlet pipe are poured in sampling water source, and the circulation that so colds and heat succeed each other is some
After secondary, developing result is preferable.In addition, in order to obtain more preferable developing result, can also be selectively turned on during flushing line
One in 7th magnetic valve 28, the 8th magnetic valve 29, selective flushing is carried out to pipeline.
The bottom of the flow cytometer is provided with exhausted bath box 31.Detection waste liquid automatic can be discharged into useless by the flow cytometer
In liquid case 31.
Embodiment 2
A kind of online test method of algae on-line detecting system, comprises the following steps：
1) sample：Selectively turn on the second magnetic valve 7 being connected with the second water inlet pipe 4, the 4th magnetic valve 10 and compacted
Dynamic pump 9, closes other magnetic valves, and the water source sample in water inlet pipe collection sampling water source, Mei Geyi are controlled by the peristaltic pump 9
The section time pumps quantitative water source sample into the pond, and acquisition rate is 2L/min, and the sample water source passes through silt mistake
Filter 23 filters silt；
2) pre-treatment：When the water source sample reaches 3L, stop pumping water source and start pretreatment, first turn on terminal electricity
The control software of brain 35, sets the 3000 turns/min of rotating speed of the magnetic stirring apparatus 12；Control the ultrasonic cell disintegration instrument 13
Power be 300W, the ultrasonic cell disintegration instrument 13 handles 33 circulations, and each circulation includes ultrasonic vibration 10s, stopping
15s；
The water source sample that the magnetic stirring apparatus 12 is pointed in pond 11 is stirred, to ensure the uniform of sample water source
Property, avoid algae from floating accuracy that the follow-up CytoSense flow cytometers 17 of influence detect；The ultrasonic cell disintegration instrument 13
Ultrasonic vibration is carried out to the water source sample in pond 11 by coupled ultrasonic transformer 15, fully dispersed algae is reached especially
The purpose of common Microcystis aeruginosa (connected and finally flocked together into bulk by ICC between alga cells) in the body of lake, to the greatest extent
The big group of algae may be broken up as groupuscule even monomer.
3) detect：When the pretreatment process is completed, by the control software of terminal computer 35, set described
CytoSense flow cytometers 17 extract the sample water source for passing through pretreatment by 2 μ l/s of pump speed out of described pond 11, simultaneously
Start detection, the sheath fluid chooses pure water, the sample water source of extraction is completed the number of algae, alga cells length, size,
The parameter detectings such as form, granularity, pigment, peak number, algae group character simultaneously draw corresponding pulse collection of illustrative plates.
Embodiment 3
A kind of online test method of algae on-line detecting system, comprises the following steps：
1) sample：Selectively turn on the 3rd magnetic valve 8 being connected with the 3rd water inlet pipe 5, the 4th magnetic valve 10 and compacted
Dynamic pump 9, closes other magnetic valves, and the water source sample in water inlet pipe collection sampling water source, Mei Geyi are controlled by the peristaltic pump 9
The section time pumps quantitative water source sample into the pond, and acquisition rate is 0.5L/min, and the sample water source passes through silt
Filter 23 filters silt；
2) pre-treatment：When the water source sample reaches 1L, stop pumping water source and start pretreatment, first turn on terminal electricity
The control software of brain 35, sets the 1000 turns/min of rotating speed of the magnetic stirring apparatus 12；Control the ultrasonic cell disintegration instrument 13
Power be 660W, the ultrasonic cell disintegration instrument 13 handles 10 circulations, and each circulation includes ultrasonic vibration 7s, stopping 10s；
The water source sample that the magnetic stirring apparatus 12 is pointed in pond 11 is stirred, to ensure the uniform of sample water source
Property, avoid algae from floating accuracy that the follow-up CytoSense flow cytometers 17 of influence detect；The ultrasonic cell disintegration instrument 13
Ultrasonic vibration is carried out to the water source sample in pond 11 by coupled ultrasonic transformer 15, fully dispersed algae is reached especially
The purpose of common Microcystis aeruginosa (connected and finally flocked together into bulk by ICC between alga cells) in the body of lake, to the greatest extent
The big group of algae may be broken up as groupuscule even monomer.
3) detect：When the pretreatment process is completed, by the control software of terminal computer 35, set described
CytoSense flow cytometers 17 extract the sample water source for passing through pretreatment by 0.5 μ l/s of pump speed out of described pond 11, together
When start detection, the sheath fluids of the CytoSense flow cytometers 17 chooses pure water, and algae is completed to the sample water source of extraction
The parameter detecting such as number, alga cells length, size, form, granularity, pigment, peak number, the special feature of algae group and draw corresponding
Pulse collection of illustrative plates.
More than, it is only the present invention preferably embodiment, but the protection domain of invention is not limited thereto, and it is any ripe
Know those skilled in the art the invention discloses technical scope in, the change or replacement that can be readily occurred in should all be contained
Cover within protection scope of the present invention.Therefore, protection scope of the present invention should be defined by the protection domain of claims.
Claims (10)
1. a kind of algae on-line detecting system, including water inlet pipe, one end of the water inlet pipe are stretched into collection water sample, the water inlet
The other end of pipe is connected with pond, and magnetic valve and use are connected with turn on the water inlet pipe between collection water sample and the pond
In the water pump of sampling, the bottom in the pond is provided with magnetic stirring apparatus, and the side in the pond is provided with ultrasonic cell disintegration
Instrument, the ultrasonic cell disintegration instrument is connected by cable with one end of ultrasonic transformer, and the other end of the ultrasonic transformer stretches into the water
In pond, level sensor is provided with the pond, the pond is flowed by sample feeding pipe with the CytoSense for algae detection
Formula cell instrument is connected, and the sidewall bottom in the pond is connected with drainpipe, and the side wall upper part in the pond is connected with overflow pipe；
The magnetic stirring apparatus, ultrasonic cell disintegration instrument, CytoSense flow cytometers, level sensor, magnetic valve, water pump difference
Electrically connected with electronic control box, the electronic control box is electrically connected with terminal computer.
2. a kind of online test method of algae on-line detecting system according to claim 1, it is characterised in that including such as
Lower step：
1) sample：Open the control software of terminal computer, the magnetic valve and water pump are opened in control, by the water pump control into
Water source sample in water pipe collection sampling water source, quantitative water source sample is pumped into the pond, is gathered at regular intervals
Speed is 0.5-2L/min；
2) pre-treatment：When the water source sample reaches predetermined amount, by the control software of terminal computer, the magnetic force is set
Agitator, the relevant parameter of ultrasonic cell disintegration instrument and operation, the terminal computer send electric signal to the electronic control box,
The electronic control box controls magnetic stirring apparatus, ultrasonic cell disintegration instrument to be operated according to default parameter respectively, and the magnetic force is stirred
Mix the water source sample that device is pointed in pond to be stirred, it is ensured that the uniformity at sample water source, avoid algae from floating；The ultrasound
Cell crushing instrument carries out ultrasonic vibration by coupled ultrasonic transformer to the water source sample in pond, reaches fully dispersed algae
Purpose；
3) detect：When the pretreatment process is completed, by the control software of terminal computer, the CytoSense streams are controlled
Formula cell instrument is brought into operation, and the CytoSense flow cytometers can be extracted by pre- by regular time and quantity out of described pond
The sample water source of processing, and the sample water source of extraction is completed the number of algae, alga cells length, size, form, granularity,
Pigment, peak number, algae group character detect and draw corresponding pulse collection of illustrative plates.
3. the online test method of algae on-line detecting system according to claim 2, it is characterised in that the water pump is
Peristaltic pump；The bifurcated at one end that the water inlet pipe stretches into sample water source forms multiple water inlet pipes, the magnetic valve have it is multiple, it is many
The individual water inlet pipe is connected by a magnetic valve with the water inlet pipe respectively；Multiple water inlet pipes are respectively protruding into described
At the different depth of sample water source vertical direction.
4. the online test method of algae on-line detecting system according to claim 3, it is characterised in that the water inlet pipe
Stretch into the first water inlet pipe of bifurcated at one end formation, the second water inlet pipe and the 3rd water inlet pipe at sample water source, the magnetic valve
Including the first magnetic valve, the second magnetic valve, the 3rd magnetic valve and the 4th magnetic valve, first water inlet pipe, the second water inlet branch
Pipe and the 3rd water inlet pipe are connected by the first magnetic valve, the second magnetic valve, the 3rd magnetic valve with water inlet pipe respectively, the water inlet
Pipe is connected with the 4th magnetic valve and silt filter positioned at one section between peristaltic pump and pond；First water inlet pipe is located at
Sample water source bottom 0.5m, second water inlet pipe is located at sample water source bottom 1m, and the 3rd water inlet pipe is located at sample
Water source bottom 1.5m.
5. the online test method of algae on-line detecting system according to claim 4, it is characterised in that in sampling step
In, selectively turn on one or several and the 4th electromagnetism in first magnetic valve, the second magnetic valve, the 3rd magnetic valve
Valve, can realize the detection of algae at the different depth to sample water source vertical direction.
6. the online test method of algae on-line detecting system according to claim 5, it is characterised in that in preceding processing step
In rapid, start pretreatment when the pond water is 1-3L；Control the revolution of the magnetic stirring apparatus turn for 1000-3000/
min；The power for controlling the ultrasonic cell disintegration instrument is 300-660W, and the ultrasonic cell disintegration instrument handles 10-33 circulation,
Each circulation includes ultrasonic vibration 3-10s, stops 3-15s.
7. the online test method of algae on-line detecting system according to claim 6, it is characterised in that in detecting step
In, control the CytoSense flow cytometers enters pump rate for 0.5-2 μ l/s, and collecting sample water source amount is 0.1-
0.5ml, the CytoSense flow cytometer sheath fluids choose pure water.
8. the online test method of algae on-line detecting system according to claim 7, it is characterised in that the algae exists
Line detecting system also includes sound proof box, and the ultrasonic transformer, magnetic stirring apparatus and pond are located in the sound proof box.
9. the algae on-line detecting system according to claim 1 or 8, it is characterised in that the side wall upper part in the pond is also
It is connected with hot water injection's pipe, the water inlet pipe and is also associated with cold water flush pipe, hot water injection's pipe and the cold water flush
Communicating pipe is connected between pipe, hot water injection's pipe, the cold water flush pipe are in the pipe end positioned at the communicating pipe side
The 5th magnetic valve and the 6th magnetic valve are connected at mouthful, hot water injection's pipe, the cold water flush pipe are positioned at described
Communicating pipe side pipeline on be also respectively connected with control piper switch the 7th magnetic valve and the 8th magnetic valve.
10. algae on-line detecting system according to claim 7, it is characterised in that the bottom of the flow cytometer is set
There is exhausted bath box.
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