CN108107025A - A kind of water quality detection method and system - Google Patents
A kind of water quality detection method and system Download PDFInfo
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- CN108107025A CN108107025A CN201711194214.7A CN201711194214A CN108107025A CN 108107025 A CN108107025 A CN 108107025A CN 201711194214 A CN201711194214 A CN 201711194214A CN 108107025 A CN108107025 A CN 108107025A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
Abstract
The invention discloses a kind of water quality detection method and system, wherein method includes：Detected sample water is mixed with fluorescent dye, the detected sample water after being dyed；Wherein, fluorescent dye is the reagent that fluorescence is generated under predetermined wavelength laser excitation；Laser irradiation is carried out to the detected sample water after dyeing by signal inspection device, obtains scattered light intensity and/or fluorescence intensity；The testing result to sample water to be detected is obtained according to scattered light intensity and/or fluorescence intensity.Water quality monitoring method disclosed by the invention, sample water to be detected is dyed with fluorescent dye, then sample water to be detected is irradiated with laser, testing result is obtained according to the scattering light and/or fluorescence intensity of generation, due to analyzing the water quality of detected sample water by detecting scattering light and/or fluorescence intensity, energy quick obtaining testing result, saves detection time, improves detection efficiency.
Description
Technical field
The present invention relates to technical field of microbial detection, and in particular to a kind of water quality detection method and system.
Background technology
As economic rapid development, the rapid growth of population and people's material and culture and the continuous of living standard carry
Height, the whole world gradually increase the demand of water, and disparities between supply and demand become increasingly conspicuous.It is incident be water resource it is increasingly deficient with
And the water pollution problems gradually protruded.How to ensure drinking water safety is always the focus of global insider's common concern.Prestige
Coerce that the factor of drinking water safety is numerous, and it is pathogenic microorganisms health to be threatened maximum, so real time on-line monitoring is drunk
It is particularly important with content of microorganisms in water.
In existing water quality in microorganism detection technology：The inspection prepared using cold water coagulator mixed-powder and fluid nutrient medium
It surveys gel entrapment culture drinking water microbe colony and counts, although shortening detection time, visible colonies can only be detected.Using point
PCR in sub- biology realizes drinking water Escherichia coli molecular Biological Detection and utilizes RNA molecule
In the presence of characterizing whether drinking water or food are polluted by pathogenic bacteria, both molecular biology methods can only detect known a certain
Class or a few quasi-microorganisms, and microbe species present in water body are various, this just need simultaneously to all kinds of common bacteriums into
Row detection, the limitation of the above method considerably increase the workload and cost of detection, are not suitable for drinking water safety production
Quick detection.
Total plate count in sample is calculated by ATP contents in test sample microorganism indirectly；Using in microorganism
There are different colours with chromogenic substrate reaction in culture medium in enzyme-specific, then extrapolates containing for microorganism by spectrophotometer
Amount；Utilize the relation of biodegradable organic and the metabolism of the latter and dissolved oxygen in microorganism growth rate in water and water
The correspondence of consumption assesses Source water and drinking water microbial contamination risk.These above-mentioned method sensitivity are not high enough；It surveys
Determine easily by the interference of ion, salinity, pH value, free state ATP, enzymatic activity and environment temperature etc.；It is thin population of samples can only to be provided
Bacterium is horizontal；By the indirectly testing means bacterial population that draws of conversion and actual content there are certain deviation, and conversion relation must
Go out to need very big workload.
The content of the invention
In view of this, an embodiment of the present invention provides a kind of water quality detection method and system, to solve to each in drinking water
The problem of detection speed is slow, testing efficiency is low when quasi-microorganism or other particles are detected.
First aspect present invention provides a kind of water quality detection method, including：
Detected sample water is mixed with fluorescent dye, the detected sample water after being dyed；Wherein, fluorescent dye is
The reagent of fluorescence is generated under predetermined wavelength laser excitation；By signal inspection device to the detected sample water after dyeing into
Row laser irradiates, and obtains scattered light intensity and/or fluorescence intensity；It is obtained according to scattered light intensity and/or fluorescence intensity to be checked
The testing result of sample water.
Optionally, the testing result of sample water to be detected is included according to scattered light intensity and/or fluorescence intensity acquisition：
Microbial cell and non-microorganism particle are identified according to the scatter diagram of scattered light intensity and fluorescence intensity.
Optionally, when signal inspection device is the laser of 488nm wavelength, according to scattered light intensity and fluorescence intensity
Scatter diagram identify that microbial cell and non-microorganism cell include：
The particle of only 488nm scattering light is determined as non-microorganism particle.
Optionally, microbial cell and non-microorganism particle are identified according to the scatter diagram of scattered light intensity and fluorescence intensity
Afterwards, further include：
Micro-organisms living cell and microorganism damaged cell are identified according to the fluorescence intensity of microbial cell.
Optionally, when signal inspection device is the laser of 488nm wavelength, according to the fluorescence intensity of microbial cell
Identify that micro-organisms living cell and microorganism damaged cell include：
Fluorescence intensity will be inspired more than first threshold, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, but
Fluorescence intensity cannot be inspired more than second threshold, wavelength is 560nm~720nm, and wave crest is determined as the particle of 617nm fluorescence
Micro-organisms living cell；
Fluorescence intensity will be inspired more than the 3rd threshold value, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, together
When can also inspire fluorescence intensity more than the 4th threshold value, wavelength is 560nm~720nm, and wave crest determines for the particle of 617nm fluorescence
For microorganism damaged cell；And/or fluorescence intensity can only be inspired more than the 5th threshold value, wavelength is 560nm~720nm, ripple
Peak is determined as microorganism damaged cell for the particle of 617nm fluorescence.
Optionally, according to the fluorescence intensity of microbial cell identify micro-organisms living cell and microorganism damaged cell it
Afterwards, further include：
According to the scattered light intensity of microbial cell and the scatter diagram of fluorescence intensity detection micro-organisms living cell quantity and
Microorganism damaged cell quantity.
Optionally, micro-organisms living cell is detected according to the scattered light intensity of microbial cell and the scatter diagram of fluorescence intensity
After quantity and microorganism damaged cell quantity, microbial cell concentration is calculated using following formula：
ρi=ni/(V·w)；
Wherein, ρiRepresent microbial cell concentration, niRepresent the quantity of micro-organisms living cell and microorganism damaged cell, V tables
Show dilution after detected sample water volume, w represent dilute sample in detected sample water volume with dilute after it is to be detected
The ratio of the volume of sample water.
Optionally, fluorescent dye includes the first fluorescent dye and the second fluorescent dye；Wherein, the first fluorescent dye use can
The cell membrane of living cells is permeated, and has the reagent of compatibility with DNA in cell；Second fluorescent dye uses can only be by imperfect
Dead cell or be damaged the cell membrane of cell, and have the reagent of compatibility with DNA in cell.
Optionally, the first fluorescent dye is SYBR Green I；Second fluorescent dye is Presidium Iodide.
Optionally, detected sample water with fluorescent dye is mixed, before the detected sample water after being dyed, also wrapped
It includes：
Using dilution reagent dilutions detected sample water；
Reagent, detected sample water and fluorescent dye mixing will be diluted using oscillator, obtain the solution of mixing.
Optionally, it is distilled water and/or deionized water to dilute reagent.
Optionally, method further includes：
Temperature is used to be protected from light the solution for cultivating mixing for 28 DEG C to 40 DEG C of constant temperature incubator.
Optionally, before using distilled water and/or deionized water dilution detected sample water, further include：
Filter the impurity in detected sample water.
Water quality monitoring method disclosed in first aspect present invention dyes sample water to be detected with fluorescent dye, and
Sample water to be detected is irradiated with laser afterwards, testing result is obtained according to the scattering light and/or fluorescence intensity of generation, due to
The water quality of detected sample water is analyzed by detecting scattering light and/or fluorescence intensity, energy quick obtaining testing result saves inspection
The time is surveyed, improves detection efficiency.
Second aspect of the present invention provides a kind of water quality detection system, including：
Sample treatment modules, for detected sample water to be mixed with fluorescent dye, the detected sample after being dyed
Water；Wherein, fluorescent dye is the reagent that fluorescence is generated under predetermined wavelength laser excitation；
Detection module carries out laser irradiation to the detected sample water after dyeing for passing through signal inspection device, obtains
Take scattered light intensity and/or fluorescence intensity；
Analysis module, for obtaining the testing result to sample water to be detected according to scattered light intensity and/or fluorescence intensity.
Optionally, signal inspection device includes：Sequentially connected laser and exciting light reshaper；Wherein, exciting light
Reshaper is used to circular laser facula being shaped as with detecting the comparable oval shaping light of particle size in detected sample water
Spot.
Optionally, detection module further includes：First detector, the second detector, the 3rd detector, the 4th detector and stream
Dynamic room；Wherein, detected sample water is housed in flow chamber；First detector be used to detecting microorganism in detected sample water by
The fluorescence that damage cell is sent by shaping hot spot；Second detector be used to detecting micro-organisms living cell in detected sample water or
The fluorescence that person's damaged cell is sent by shaping hot spot；The biologic grain that 3rd detector is used to detect in detected sample water leads to
Cross the side scattered light that shaping hot spot is sent；The biologic grain that 4th detector is used to detect in detected sample water passes through shaping
The forward scattering light that hot spot is sent.
Optionally, one or more two points of mirrors and one or more are included between the first detector and shaping hot spot
A band pass filter；Between second detector and shaping hot spot include one or more two points of mirrors and one or more
Band pass filter；Include one or more two points of mirrors and one or more band between 3rd detector and shaping hot spot
Pass filter piece.
Optionally, two points of mirrors divide mirror for high anti-low pass two.
Optionally, high anti-low pass two divides the reflection wavelength of mirror to be 560nm~720nm, and transmission peak wavelength is less than 560nm；Alternatively,
High anti-low pass two divides the reflection wavelength of mirror, and for 490nm~550nm, transmission peak wavelength is less than 490nm.
Optionally, the first detector is avalanche diode or photomultiplier；Second detector is avalanche diode or light
Electric multiplier tube；3rd detector is avalanche diode or photomultiplier；4th detector is photodiode.
Optionally, analysis module is additionally operable to according to the first detector, the second detector, the 3rd detector and the 4th detector
The scatter diagram for collecting scattered light intensity and fluorescence intensity identifies microbial cell and non-microorganism particle.
Optionally, when signal inspection device is the laser of 488nm wavelength, analysis module is additionally operable to will be only the
Three detectors and the 4th detector detect that the particle during scattering light of 488nm wavelength is determined as non-microorganism particle.
Optionally, when signal inspection device is the laser of 488nm wavelength, analysis module is additionally operable to：
Fluorescence intensity will be inspired more than first threshold, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, but
Fluorescence intensity cannot be inspired more than second threshold, wavelength is 560nm~720nm, and wave crest is determined as the particle of 617nm fluorescence
Micro-organisms living cell；
Fluorescence intensity will be inspired more than the 3rd threshold value, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, together
When can also inspire fluorescence intensity more than the 4th threshold value, wavelength is 560nm~720nm, and wave crest determines for the particle of 617nm fluorescence
For microorganism damaged cell；And/or fluorescence intensity can only be inspired more than the 5th threshold value, wavelength is 560nm~720nm, ripple
Peak is determined as microorganism damaged cell for the particle of 617nm fluorescence.
Optionally, analysis module is additionally operable to be detected according to the scattered light intensity of microbial cell and the scatter diagram of fluorescence intensity
The quantity of micro-organisms living cell and microorganism damaged cell quantity.
Optionally, fluorescent dye includes the first fluorescent dye and the second fluorescent dye；Wherein, the first fluorescent dye use can
The cell membrane of living cells is permeated, and has the reagent of compatibility with DNA in cell；Second fluorescent dye uses can only be by imperfect
Dead cell or be damaged the cell membrane of cell, and have the reagent of compatibility with DNA in cell.
Optionally, the first fluorescent dye is SYBR Green I；Second fluorescent dye is Presidium Iodide.
Optionally, sample treatment modules further include：First constant displacement pump, the second constant displacement pump, the 3rd constant displacement pump and the 4th are quantitative
Pump；
First constant displacement pump extracts detected sample water after the filtering of the first setting capacity to mixer from sample bottle
In；Second constant displacement pump extracts the dilution reagent of the second setting capacity into mixer from dilution reagent fluid reservoir；3rd is quantitative
The first fluorescent dye that the 3rd setting capacity is extracted from the first fluorescent dye fluid reservoir is pumped into mixer；4th constant displacement pump from
The second fluorescent dye of the 4th setting capacity is extracted in second fluorescent dye fluid reservoir into mixer.
Optionally, the first constant displacement pump, the second constant displacement pump, the 3rd constant displacement pump and the 4th constant displacement pump are plunger pump or syringe pump.
Optionally, sample treatment modules further include：First solenoid valve, second solenoid valve, the 5th constant displacement pump and the 6th are quantitative
Pump；
Second solenoid valve connects the pipeline of the pipeline of the sample bottle equipped with detected sample water and the 5th constant displacement pump, carries out
Quantitative extraction to sample water to be detected, and the detected sample water of extraction is delivered to mixer, alternatively, second solenoid valve will
The pipeline of the pipeline and the 5th constant displacement pump that dilute reagent fluid reservoir is connected, and is carried out the quantitative extraction to diluting reagent, and will be extracted
Dilution reagent be delivered to mixer；
Second solenoid valve connects the pipeline of the pipeline of the first fluorescent dye fluid reservoir and the 6th constant displacement pump, carries out to first
The quantitative extraction of fluorescent dye, and the first fluorescence of extraction dye is delivered to mixer, alternatively, second solenoid valve is by the second fluorescence
The pipeline of the pipeline of dyestuff fluid reservoir and the 6th constant displacement pump is connected, and is carried out the quantitative extraction to the second fluorescent dye, and will be extracted
The second fluorescent dye be delivered to mixer.
Optionally, the 5th constant displacement pump and the 6th constant displacement pump are plunger pump or syringe pump；First solenoid valve and second solenoid valve
For two-bit triplet solenoid valve.
Optionally, it is distilled water and/or deionized water to dilute reagent.
Optionally, mixer includes oscillator, and oscillator contaminates for that will dilute reagent, detected sample water, the first fluorescence
Material and the second fluorescent dye mixing, obtain the solution of mixing.
Optionally, mixer is connected with the constant temperature incubator that temperature is 28 DEG C to 40 DEG C.
Optionally, it is provided with the first negative pressure sampling pump between flow chamber and mixer, the first negative pressure sampling pump is for from mixed
Clutch extracts mixed solution.
Optionally, sample treatment modules further include the second negative pressure sampling pump, the second negative pressure sampling pump for initial condition into
Row extracts and obtains detected sample water.
Optionally, sample treatment modules further include the 3rd solenoid valve, the 4th solenoid valve, the 3rd negative pressure sampling pump and waste liquid
Tank；
The 3rd solenoid valve is controlled to connect the pipeline of mixer and the 3rd negative pressure sampling pump line road, is sampled using the 3rd negative pressure
Solution in mixer is transported in waste liquid tank by pump；The 3rd solenoid valve is controlled to pump the pipeline of mixer and the sampling of the 3rd negative pressure
Pipe cutting after, the 4th solenoid valve of control connects the pipeline of pipeline in sample bottle and the 3rd negative pressure sampling pump, recycling the
Filtered sample in sample bottle is transported in waste liquid tank by three negative pressure sampling pump.
Water quality monitoring system disclosed in second aspect of the present invention, sample treatment modules are used for fluorescent dye to sample to be detected
Product water is dyed, and then for laser to be irradiated sample water to be detected, analysis module is used for according to production detection module
Raw scattering light and/or fluorescence intensity obtain testing result, since water quality monitoring system is strong by detecting scattering light and/or fluorescence
Spend to analyze the water quality of detected sample water, can quick obtaining testing result, save detection time, improve detection efficiency.
Description of the drawings
The features and advantages of the present invention can be more clearly understood by reference to attached drawing, attached drawing is schematically without that should manage
It solves to carry out any restrictions to the present invention, in the accompanying drawings：
Fig. 1 is a kind of flow chart of water quality detection method in the embodiment of the present invention 1；
Fig. 2 is a kind of fluorescence intensity of water quality detection method and wavelength of fluorescence distribution map in the embodiment of the present invention 1；
Fig. 3 is a kind of module diagram of water quality monitoring system in the embodiment of the present invention 2；
Fig. 4 is a kind of a kind of structure diagram of water quality monitoring system in the embodiment of the present invention 2；
Fig. 5 is a kind of a kind of structure diagram of water quality monitoring system in the embodiment of the present invention 2；
Fig. 6 is a kind of a kind of forward scattering optical detector schematic diagram of water quality monitoring system in the embodiment of the present invention 2；
Fig. 7 is a kind of a kind of sample treatment modules schematic diagram of water quality monitoring system in the embodiment of the present invention 2；
Fig. 8 is a kind of another sample treatment modules schematic diagram of water quality monitoring system in the embodiment of the present invention 2；
Fig. 9 is a kind of overall system structure schematic diagram of water quality monitoring system in the embodiment of the present invention 2；
Figure 10 is that a kind of water quality monitoring system illustrates the system structure that multiple water sources are monitored in the embodiment of the present invention
Figure；
Reference sign：
100- sample treatment modules；
The first constant displacement pumps of 101-；The second constant displacement pumps of 102-；The 3rd constant displacement pumps of 103-；The 4th constant displacement pumps of 104-；
105- sample bottles；106- mixers；107- dilutes reagent fluid reservoir；108- the first fluorescent dye fluid reservoirs；109-
Second fluorescent dye fluid reservoir；The first solenoid valves of 110-；111- second solenoid valves；The 5th constant displacement pumps of 112-；113- the 6th is quantitative
Pump；114- constant temperature incubators；The first negative pressure of 115- sampling pump；The 3rd solenoid valves of 117-；The 4th solenoid valves of 118-；119- the 3rd is negative
Pressure sampling pump；120- waste liquid tanks；
200- detection modules；
The first detectors of 201-；The second detectors of 202-；The 3rd detectors of 203-；The 4th detectors of 204-；205- flows
Room；206- bis- divides mirror；207- band pass filters；208- bis- divides mirror；209- band pass filters；210- band pass filters；
300- signal inspection devices；
301- lasers；302- exciting light reshapers；
400- analysis modules.
Specific embodiment
To make the purpose, technical scheme and advantage of the embodiment of the present invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, the technical solution in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is
Part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those skilled in the art are not having
All other embodiments obtained under the premise of creative work are made, belong to the scope of protection of the invention.
Embodiment 1
The present embodiment provides a kind of water quality detection method, as shown in Figure 1, the flow includes the following steps：
Step S1 mixes detected sample water with fluorescent dye.Detected sample water after being dyed；Wherein, it is glimmering
Photoinitiator dye is the reagent that fluorescence is generated under predetermined wavelength laser excitation.Specifically, fluorescent dye has the DNA of cell affine
Property, can be embedded into DNA double chain or can with DNA covalent bonds or can physical absorption on some group of cell.
Step S2 carries out laser irradiation by signal inspection device to the detected sample water after dyeing.Specific real
It applies in example, sample water to be detected is irradiated with laser, when there is dyed particles to pass through, dyed particles can inspire glimmering
Light；Scattering light can be only generated when the particle that is unstained is irradiated through laser.
Step S3 obtains scattered light intensity and/or fluorescence intensity.In a particular embodiment, particle is detected by detector
The scattering light and/or fluorescence sent.
Step S4 obtains the testing result to sample water to be detected according to scattered light intensity and/or fluorescence intensity.Specific
In embodiment, the situation of scattering light and/or fluorescence is generated according to particle to differentiate that particle is non-microorganism particle or microorganism
Cell；Differentiate that microbial cell is specifically competent cell or damaged cell according to fluorescence intensity (including dead cell).
As optional embodiment, the inspection to sample water to be detected is obtained according to scattered light intensity and/or fluorescence intensity
Surveying result includes：Microbial cell and non-microorganism particle are identified according to the scatter diagram of scattered light intensity and fluorescence intensity.
In specific embodiment, when only scattering light in scatter diagram, which is non-microorganism particle；When existing scattering light in scatter diagram
When having fluorescence again, which is microbial cell.
As optional embodiment, when signal inspection device is the laser of 488nm wavelength, according to scattered light intensity
Degree and the scatter diagram of fluorescence intensity identify that microbial cell and non-microorganism cell include the particle that will there was only 488nm scattering light
It is determined as non-microorganism particle.In a particular embodiment, fluorescent dye does not have non-microorganism particle affinity, therefore non-micro- life
Composition granule will not inspire fluorescence through laser irradiation.Under the laser irradiation that wavelength is 488nm, non-microorganism particle is only capable of generating
The scattering light of 488nm.
As optional embodiment, according to the scatter diagram of scattered light intensity and fluorescence intensity identify microbial cell and
After non-microorganism particle, further include：According to the fluorescence intensity of microbial cell identify micro-organisms living cell and microorganism by
Damage cell.In a particular embodiment, since micro-organisms living cell is different with the permeability of cell membrane of microorganism damaged cell, some
Fluorescent dye, such as propidium iodide (Presidium Iodide, PI), it is impossible to by the cell membrane of micro-organisms living cell, be only capable of
By the cell membrane of microorganism damaged cell, so as to carry out fluorescent staining to the DNA of microorganism damaged cell；And some fluorescence contaminate
Material, such as SYBR Green I can carry out fluorescent staining by the cell membrane of micro-organisms living cell to its DNA.Therefore, using
In the case that this two fluorochrome carries out fluorescent staining to sample to be tested water, micro-organisms living cell and microorganism damaged cell
For DNA under laser irradiation, the fluorescence intensity inspired is different.
It is thin according to microorganism when signal inspection device is the laser of 488nm wavelength as optional embodiment
The fluorescence intensity of born of the same parents identifies that micro-organisms living cell and microorganism damaged cell include to inspire fluorescence intensity more than first
Threshold value, wavelength are 490nm~550nm, and wave crest is 521nm fluorescence, but cannot inspire fluorescence intensity more than second threshold, wavelength
For 560nm~720nm, wave crest is determined as micro-organisms living cell for the particle of 617nm fluorescence；Fluorescence intensity will be inspired to be more than
3rd threshold value, wavelength are 490nm~550nm, and wave crest is 521nm fluorescence, while can also inspire fluorescence intensity more than the 4th threshold
Value, wavelength are 560nm~720nm, and wave crest is determined as microorganism damaged cell for the particle of 617nm fluorescence；It and/or can only
Fluorescence intensity is inspired more than the 5th threshold value, wavelength is 560nm~720nm, and wave crest is determined as micro- life for the particle of 617nm fluorescence
Object damaged cell.In a particular embodiment, using both fluorescent dyes of SYBR Green I and Presidium Iodide, use
The laser of 488nm is irradiated sample to be tested water, as shown in Fig. 2, being inspired by the SYBR Green I living cells dyed
Fluorescence relative intensity for 85% or so, fluorescent wavelength ranges 490nm~550nm, wave crest 521nm；And pass through the thin of PI dyeing
The fluorescence relative intensity that born of the same parents inspire is 9% or so, fluorescent wavelength ranges 560nm~720nm, wave crest 617nm.Some microorganisms
Damaged cell or dead cell can be dyed by SYBR Green I and PI simultaneously, therefore can detect both the above varying strength simultaneously
With the fluorescence of wavelength.
As optional embodiment, micro-organisms living cell and microorganism are identified according to the fluorescence intensity of microbial cell
After damaged cell, further include：It is lived according to the scattered light intensity of microbial cell and the scatter diagram of fluorescence intensity detection microorganism
The quantity of cell and microorganism damaged cell quantity.In a particular embodiment, laser to the detected sample water of one-way flow into
Row irradiation, when there is particle to pass through, after carrying out fluorescence intensity identification to it, counts it.
It is micro- according to the scattered light intensity of microbial cell and the detection of the scatter diagram of fluorescence intensity as optional embodiment
After the quantity of living biological cell and microorganism damaged cell quantity, microbial cell concentration is calculated using following formula：
ρi=ni/(V·w)；
Wherein, ρiRepresent microbial cell concentration, niRepresent the quantity of micro-organisms living cell and microorganism damaged cell, V tables
Show dilution after detected sample water volume, w represent dilute sample in detected sample water volume with dilute after it is to be detected
The ratio of the volume of sample water.In a particular embodiment, the micro-organisms living cell quantity n that statistic mixed-state arrives1, microorganism is damaged carefully
The quantity n of born of the same parents' (including dead cell)2, the sample water volume V of current detection is counted, if there are diluted situations, is calculated before diluting
Sample water volume, such as dilution before sample water volume be v1, diluent volume v2, then the sample water volume V=v that detects1
+v2；
W=v1/(v1+v2)=v1/ V, so wV=(v1+v2)·v1/(v1+v2)=v1。
Micro-organisms living cell concentration ρ1=n1/v1, microorganism damaged cell (including dead cell) concentration ρ2=n2/v2。
In a specific embodiment, the concentration ρ of non-microorganism particle can also be counted3=n3/(V·w).Wherein n3To be non-
The quantity of microbe granular.
Specifically, 1 can be selected：10,1：100 or 1：1000 dilution ratio dilutes.
As optional embodiment, fluorescent dye includes the first fluorescent dye and the second fluorescent dye；Wherein, first is glimmering
Photoinitiator dye uses the cell membrane of permeable living cells, and has the reagent of compatibility with DNA in cell；Second fluorescent dye is using only
By incomplete dead cell or the cell membrane of cell can be damaged, and have the reagent of compatibility with DNA in cell.Specific real
It applies in example, while is mixed using two kinds of fluorescent dyes and sample to be tested water, you can make the micro-organisms living cell in sample to be tested water
It is all colored with the DNA of microorganism damaged cell (including dead cell).
As optional embodiment, the first fluorescent dye is SYBR Green I；Second fluorescent dye is Presidium
Iodide.In a particular embodiment, SYBR Green I can penetrate the cell membrane of micro-organisms living cell, and PI is only capable of by impaired thin
Born of the same parents and/or the cell membrane of dead cell.
As optional embodiment, detected sample water is mixed with fluorescent dye, the test sample to be checked after being dyed
Before product water, further include using dilution reagent dilutions detected sample water；Reagent, detected sample water will be diluted using oscillator
And fluorescent dye mixing, obtain the solution of mixing.In a particular embodiment, if detected sample water middle particle concentration is excessively high,
Then it is diluted, reduces granule density, during ensureing that detected sample water is flowed through at laser irradiation in detection process, every time
Only pass through there are one particle.
As optional embodiment, dilution reagent is distilled water and/or deionized water.In a particular embodiment, with nothing
The water body of other microbial cells and non-microorganism particle makees diluent so that testing result influences from diluent.
As optional embodiment, temperature is used to be protected from light for 28 DEG C to 40 DEG C of constant temperature incubator and cultivates the molten of mixing
Liquid.In a particular embodiment, in order to improve the reaction effect of filtered sample and fluorescent dye, constant temperature incubator uses and is protected from light structure
Or coating is protected from light, prevent the mixed solution in ambient light irradiation constant temperature incubator.It is shown according to test result, detected sample water
Optimal reaction temperature with fluorescent dye is at 28 DEG C to 40 DEG C；Constant temperature cultivates the time in 15min to 30min.Optionally implementing
In example, thermometer is installed in constant temperature incubator, for monitoring the temperature of the mixed solution in constant temperature incubator；Using electrically heated rod
Water bath with thermostatic control is carried out to the mixed solution in mixer (106)；Or mixer (106) mixed solution is carried out using heat gun
The empty bath of constant temperature.
As optional embodiment, before distilled water and/or deionized water dilution detected sample water, further include
Filter the impurity in detected sample water.In a particular embodiment, be filtered to remove large granular impurity, including visible foreign and
Visible colonies group.
Water quality monitoring method disclosed in the present embodiment dyes sample water to be detected with fluorescent dye, then with sharp
Light is irradiated sample water to be detected, testing result is obtained according to the scattering light and/or fluorescence intensity of generation, due to passing through inspection
Scattering light and/or fluorescence intensity are surveyed to analyze the water quality of detected sample water, energy quick obtaining testing result, when saving detection
Between, improve detection efficiency.
Embodiment 2
As shown in figure 3, present embodiments providing a kind of water quality detection system, including sample treatment modules (100), mould is detected
Block (200), signal inspection device (300) and analysis module (400), wherein：
Sample treatment modules (100) are to be detected after being dyed for detected sample water to be mixed with fluorescent dye
Sample water；Wherein, fluorescent dye is the reagent that fluorescence is generated under predetermined wavelength laser excitation；Detection module (200) is used to lead to
It crosses signal inspection device (300) and laser irradiation is carried out to the detected sample water after dyeing, obtain scattered light intensity and/or glimmering
Luminous intensity；Analysis module (400) is used to obtain the detection knot to sample water to be detected according to scattered light intensity and/or fluorescence intensity
Fruit.In a particular embodiment, sample treatment modules (100) are dyed blended by fluorescent dye and detected sample water, then detect
Module (200) is irradiated sample water to be detected with laser, and the particle in detected sample water passes through signal inspection device
(300) send scattering light and/or fluorescence during laser-irradiated domain, analysis module (400) be used for according to the scattering light of generation and/
Or fluorescence intensity obtains testing result.
As optional embodiment, signal inspection device (300) includes sequentially connected laser (301) and excitation
Light shaping device (302)；Wherein, exciting light reshaper (302) be used for by circular laser facula be shaped as in detected sample water
Detect the comparable oval shaping hot spot of particle size.In a particular embodiment, exciting light reshaper (302) is by laser
(301) the circular laser facula sent is shaped as with detecting the comparable oval shaping light of particle size in detected sample water
Spot, to laser power density of the booster action on detection particle.It is realized for example with bicylindrical mirror or poly-lens etc. sharp
Light focuses on.
As optional embodiment, detection module (200) further includes the first detector (201), the second detector
(202), the 3rd detector (203), the 4th detector (204) and flow chamber (205)；Wherein, equipped with to be checked in flow chamber (205)
Sample water；The microorganism damaged cell that first detector (201) is used to detect in detected sample water is sent out by shaping hot spot
The fluorescence gone out；The micro-organisms living cell or damaged cell that second detector (202) is used to detect in detected sample water pass through
The fluorescence that shaping hot spot is sent；The biologic grain that 3rd detector (203) is used to detect in detected sample water passes through shaping light
The side scattered light that spot is sent；The biologic grain that 4th detector (204) is used to detect in detected sample water passes through shaping light
The forward scattering light that spot is sent.As shown in Figure 4 and Figure 5, when being irradiated using the laser of 488nm, the first detector (201) is used to visit
Survey wavelength be 560nm~720nm, wave crest be 617nm fluorescence, the second detector (202) for detect wavelength for 490nm~
550nm, wave crest are 521nm fluorescence, and the 3rd detector (203) is used for the side scattered light of a length of 488nm of probing wave；4th detection
Device (204) is used for the forward scattering light of a length of 488nm of probing wave.
As optional embodiment, one or more two points are included between the first detector (201) and shaping hot spot
Mirror (206) and one or more band pass filter (207)；Between second detector and shaping hot spot include one or
Multiple two points of mirrors (208) and one or more band pass filter (209)；Between 3rd detector (203) and shaping hot spot
Including one or more two points of mirrors (208) and one or more band pass filter (210).In a particular embodiment, such as
Shown in Fig. 4 and Fig. 5, multiple band pass filters are used to making the light of selected wave band to pass through and the light cut-off beyond passband；Two points of mirrors are used
In the light of reflection high band, the light of low band is transmitted.Such as：The light that two points of mirror (206) reflection wavelengths are 560nm~720nm, thoroughly
Light of the ejected wave length less than 560nm；The light that two points of mirror (207) reflection wavelengths are 490nm~550nm, transmission peak wavelength is less than 490nm's
Light.
As optional embodiment, two points of mirrors divide mirror for high anti-low pass two.
As optional embodiment, high anti-low pass two divides the reflection wavelength of mirror, and for 560nm~720nm, transmission peak wavelength is small
In 560nm；Alternatively, high anti-low pass two divides the reflection wavelength of mirror, for 490nm~550nm, transmission peak wavelength is less than 490nm.Specific
In embodiment, multiple high anti-low passes two divide the reflection wavelength of mirror and transmission peak wavelength to differ.
As optional embodiment, the first detector (201) is avalanche diode or photomultiplier；Second detector
(202) it is avalanche diode or photomultiplier；3rd detector (203) is avalanche diode or photomultiplier；4th visits
It is photodiode to survey device (204).In a particular embodiment, because non-microorganism particle and/or microbial cell are in shaping hot spot
Irradiation under the forward-scattering signal that generates it is very strong, the 4th detector (204) is detected using photodiode；It generates
Side scattered light is after two two points of mirrors, an attenuated optical signal part, therefore the 3rd detector (203) uses snowslide two
Pole pipe (APD) or photomultiplier；And the fluorescence signal that microbial cell generates under the irradiation of shaping hot spot is very weak, therefore first
Detector (201) and the second detector (202) use avalanche diode or photomultiplier.
As optional embodiment, analysis module (400) is additionally operable to according to the first detector (201), the second detector
(202), the 3rd detector (203) and the 4th detector (204) collect the scatter diagram identification of scattered light intensity and fluorescence intensity
Go out microbial cell and non-microorganism particle.In a particular embodiment, analysis module (400) is to the first detector (201), second
The scattered light intensity and fluorescence intensity that detector (202), the 3rd detector (203) and the 4th detector (204) detect summarize
Scatter diagram is formed, identifies microbial cell and non-microorganism particle.
In a specific embodiment, as shown in fig. 6, particle is irradiating-measurement zone (ellipse light spot by laser in drinking water
Area) when, it can be to space in 360 ° of solid angle direction scattering lights, the size of scattered light signal and particle, shape, plasma membrane and thin
The refractive index of intracellular grain structure is all related.
Forward scattering optical detector (SSC detectors) testing result mainly reflects that particle in drinking water is (thin comprising microorganism
Born of the same parents, sand grains, other granular debris etc.) volume size, the size of the value and the diameter of particle are near linear relation.Namely
It says, the volume size of particle is smaller, and the forward-scattering signal intensity generated under the irradiation of laser is weaker, otherwise also
So.
Lateral scattering optical detector testing result mainly reflects the internal particle architecture quality size of particle in drinking water, side
, the signal strength almost quality with intracellular granular structure more sensitive to the refraction of cell membrane, kytoplasm and nuclear membrane to scattering light
In near linear relation, that is to say, that intracellular granular structure is more complicated, and quality is bigger, and lateral scattering optical signal is stronger, instead
It is as the same.
As optional embodiment, when signal inspection device (300) is the laser of 488nm wavelength, mould is analyzed
Block (200) is additionally operable to only detect the scattering light of 488nm wavelength in the 3rd detector (203) and the 4th detector (204)
When particle be determined as non-microorganism particle.In a particular embodiment, because non-microorganism particle is without fluorescent dyeing, warp
Laser facula irradiation will not inspire fluorescence, only can send scattering light.Therefore, the laser radiation situation of wavelength 488nm is being used
Under, when only the 3rd detector (203) and the 4th detector (204) detect the scattering light of wavelength 488nm, analysis module
(400) it is non-microorganism particle to determine the detection particle.
As optional embodiment, when signal inspection device (300) is the laser of 488nm wavelength, mould is analyzed
Block (200) is additionally operable to inspire fluorescence intensity more than first threshold, and wavelength is 490nm~550nm, and wave crest is glimmering for 521nm
Light, but fluorescence intensity cannot be inspired more than second threshold, wavelength is 560nm~720nm, and wave crest is the particle of 617nm fluorescence
It is determined as micro-organisms living cell；Fluorescence intensity will be inspired more than the 3rd threshold value, wavelength is 490nm~550nm, and wave crest is
521nm fluorescence, while fluorescence intensity can be also inspired more than the 4th threshold value, wavelength is 560nm~720nm, and wave crest is glimmering for 617nm
The particle of light is determined as microorganism damaged cell；And/or fluorescence intensity can only be inspired more than the 5th threshold value, wavelength is
560nm~720nm, wave crest are determined as microorganism damaged cell for the particle of 617nm fluorescence.In a particular embodiment, use
Both fluorescent dyes of SYBR Green I and Presidium Iodide, the laser pair of laser (301) generation wavelength 488nm
Sample to be tested water is irradiated, as shown in Fig. 2, the fluorescence relative intensity inspired by the SYBR Green I living cells dyed
For 85% or so, fluorescent wavelength ranges 490nm~550nm, wave crest 521nm, fluorescence letter is detected by the second detector (202)
Number；And the fluorescence relative intensity that goes out of cell activation for passing through PI dyeing is 9% or so, fluorescent wavelength ranges 560nm~720nm,
Wave crest 617nm detects the fluorescence signal by the first detector (201).Some microorganism damaged cells or dead cell can be simultaneously
It is dyed by SYBR Green I and PI, therefore the first detector (201) and the second detector (202) can detect fluorescence letter simultaneously
Number.
As optional embodiment, analysis module (400) is additionally operable to according to the scattered light intensity of microbial cell and glimmering
The scatter diagram detection quantity of micro-organisms living cell and microorganism damaged cell quantity of luminous intensity.In a particular embodiment, whenever
First detector (201) and/or, the second detector (202) and/or, the 3rd detector (203) and/or, the 4th detector
(204) when detecting signal, counted.Or analysis module (400) identify the particle that detects for non-microorganism particle,
While micro-organisms living cell or microorganism damaged cell, it is counted.
As optional embodiment, fluorescent dye includes the first fluorescent dye and the second fluorescent dye；Wherein, first is glimmering
Photoinitiator dye uses the cell membrane of permeable living cells, and has the reagent of compatibility with DNA in cell；Second fluorescent dye is using only
By incomplete dead cell or the cell membrane of cell can be damaged, and have the reagent of compatibility with DNA in cell.Specific real
It applies in example, while is mixed using two kinds of fluorescent dyes and sample to be tested water, you can make the micro-organisms living cell in sample to be tested water
It is all colored with the DNA of microorganism damaged cell (including dead cell).
As optional embodiment, the first fluorescent dye is SYBR Green I；Second fluorescent dye is Presidium
Iodide.In a particular embodiment, SYBR Green I can penetrate the cell membrane of micro-organisms living cell, and PI is only capable of by impaired thin
Born of the same parents and/or the cell membrane of dead cell.
As optional embodiment, sample treatment modules (100) further include the first constant displacement pump (101), the second constant displacement pump
(102), the 3rd constant displacement pump (103) and the 4th constant displacement pump (104)；Wherein, the first constant displacement pump (101) is taken out from sample bottle (105)
The detected sample water after the filtering of the first setting capacity is taken into mixer (106)；Second constant displacement pump (102) is tried from dilution
The dilution reagent of the second setting capacity is extracted in agent fluid reservoir (107) into mixer (106)；3rd constant displacement pump (103) is from
The first fluorescent dye of the 3rd setting capacity is extracted in one fluorescent dye fluid reservoir (108) into mixer (106)；4th is quantitative
(104) are pumped from the second fluorescent dye of the 4th setting capacity of extraction in the second fluorescent dye fluid reservoir (109) to mixer (106)
In.In a particular embodiment, as shown in Fig. 7 or Fig. 8, the mud in drinking water resource or the drinking water of water factory's production process is filtered out
Sand, iron rust, suspended matter and this kind of granulometric impurity of microfibre prevent from influencing testing result, set blocking channel, damage instrument.Tool
Body, use flow cytometry microbial rapid detection module (200) optimal microorganism testing concentration range for 103~
107cells/mL, when microorganism concn is very big, it is necessary to the dilution sample to be tested water of constant gradient, such as setting 1：10,1：100 or
1：1000 dilution ratio is diluted, to improve the measuring accuracy of content of microorganisms.
As optional embodiment, the first constant displacement pump (101), the second constant displacement pump (102), the 3rd constant displacement pump (103) and
4th constant displacement pump (104) is plunger pump or syringe pump.
As optional embodiment, sample treatment modules further include：First solenoid valve (110), second solenoid valve
(111), the 5th constant displacement pump (112) and the 6th constant displacement pump (113), wherein：
Second solenoid valve (111) is by the pipeline and the 5th constant displacement pump (112) of the sample bottle (105) equipped with detected sample water
Pipeline connect, carry out the quantitative extraction to sample water to be detected, and the detected sample water of extraction be delivered to mixer
(106), alternatively, second solenoid valve (111) will dilute the pipeline of reagent fluid reservoir (107) and the pipeline of the 5th constant displacement pump (112)
It connects, carries out to diluting the quantitative extraction of reagent, and the dilution reagent of extraction is delivered to mixer (106)；
In a particular embodiment, as shown in figure 8, sample bottle (105) and diluent fluid reservoir (107) two-bit triplet electromagnetism
Valve and the 5th constant displacement pump (112) are connected, and sample water to be detected is diluted in case of need.
Second solenoid valve (111) is by the pipeline of the first fluorescent dye fluid reservoir (108) and the pipeline of the 6th constant displacement pump (113)
It connects, carries out the quantitative extraction to the first fluorescent dye, and the first fluorescence dye of extraction is delivered to mixer (106), alternatively,
Second solenoid valve (111) connects the pipeline of the pipeline of the second fluorescent dye fluid reservoir (109) and the 6th constant displacement pump (113), into
Quantitative extraction of the row to the second fluorescent dye, and the second fluorescent dye of extraction is delivered to mixer (106).
As optional embodiment, the 5th constant displacement pump (112) and the 6th constant displacement pump (113) are plunger pump or syringe pump；
First solenoid valve (110) and second solenoid valve (111) are two-bit triplet solenoid valve.
As optional embodiment, dilution reagent is distilled water and/or deionized water.
As optional embodiment, mixer (106) includes oscillator, and oscillator will be for will dilute reagent, to be detected
Sample water, the first fluorescent dye and the second fluorescent dye mixing, obtain the solution of mixing.In a particular embodiment, using shaking
Dynamic motor realizes the mixing of mixed liquor, to accelerate the anti-of detected sample water and the first fluorescent dye and second fluorescent dye
It should.
As optional embodiment, mixer (106) is arranged at the constant temperature incubator (114) that temperature is 28 DEG C to 40 DEG C
It is interior.In a particular embodiment, in order to improve the reaction effect of filtered sample and fluorescent dye, constant temperature incubator uses and is protected from light structure
Or coating is protected from light, prevent the mixed solution in ambient light irradiation constant temperature incubator.It is shown according to test result, detected sample water
Optimal reaction temperature with fluorescent dye is at 28 DEG C to 40 DEG C；Constant temperature cultivates the time in 15min to 30min.Optionally implementing
In example, thermometer is installed in constant temperature incubator, for monitoring the temperature of the mixed solution in constant temperature incubator；Using electrically heated rod
Water bath with thermostatic control is carried out to the mixed solution in mixer (106)；Or mixer (106) mixed solution is carried out using heat gun
The empty bath of constant temperature.
As optional embodiment, the first negative pressure sampling pump is provided between flow chamber (205) and mixer (106)
(115), the first negative pressure sampling pump (115) is used to extract mixed solution from mixer (106).In a particular embodiment, it is
Microbial cell or non-microorganism particle in realization detected sample water pass through shaping hot spot, the first negative pressure sampling pump one by one
(115) sampling rate should not be very big, therefore using flow in the peristaltic pump of the μ L/min of 10 μ L/min~200, diaphragm pump or electricity
Magnetic pumping.
As optional embodiment, sample treatment modules (100) further include the second negative pressure sampling pump (116), and second is negative
Pressure sampling pump (116) is used to that initial condition to be extracted to obtain detected sample water.In a particular embodiment, the second negative pressure is sampled
The operating status for pumping (116) is controlled by the analysis module (400) of host computer, and sampling quantity is controlled by liquid level gauge, after filtering
Detected sample water is stored by sample bottle (105).In sampling process, excluded using the gas vent on sample bottle (105) on lid
Air in sample bottle (105), and using the fluid level condition of detected sample water in liquid level gauge monitoring sample bottle (105), work as liquid
When position reaches preset value, analysis module (400) controls the second negative pressure sampling pump (116) out of service.
As optional embodiment, sample treatment modules (100) further include the 3rd solenoid valve (117), the 4th solenoid valve
(118), the 3rd negative pressure sampling pump (119) and waste liquid tank (120), wherein：
The 3rd solenoid valve (117) is controlled to connect the pipeline of mixer (106) and the 3rd negative pressure sampling pump (119) pipeline,
The solution in mixer (106) is transported in waste liquid tank (120) using the 3rd negative pressure sampling pump (119)；Control the 3rd electromagnetism
Valve (117) controls the 4th solenoid valve by after the pipe cutting of the pipeline of mixer (106) and the 3rd negative pressure sampling pump (119)
(118) pipeline of pipeline in sample bottle (105) and the 3rd negative pressure sampling pump (119) is connected, recycles the 3rd negative pressure sampling pump
(119) filtered sample in sample bottle (105) is transported in waste liquid tank (120).In a particular embodiment, sample is evacuated to improve
The efficiency of solution in product bottle (105) and mixer (106), the 3rd negative pressure sampling pump (119) preferably flow is more than 100mL/min's
Pump.
Although being described in conjunction with the accompanying the embodiment of the present invention, those skilled in the art can not depart from the present invention
Spirit and scope in the case of various modification can be adapted and modification, such modifications and variations are each fallen within by appended claims institute
Within the scope of restriction.
Claims (36)
1. a kind of water quality detection method, which is characterized in that including：
Detected sample water is mixed with fluorescent dye, the detected sample water after being dyed；Wherein, the fluorescent dye is
The reagent of fluorescence is generated under predetermined wavelength laser excitation；
Laser irradiation is carried out to the detected sample water after the dyeing by signal inspection device, obtains scattered light intensity
And/or fluorescence intensity；
The testing result to the detected sample water is obtained according to the scattered light intensity and/or fluorescence intensity.
2. water quality detection method according to claim 1, which is characterized in that according to the scattered light intensity and/or fluorescence
Intensity acquisition includes the testing result of the detected sample water：
Microbial cell and non-microorganism particle are identified according to the scatter diagram of the scattered light intensity and fluorescence intensity.
3. water quality detection method according to claim 2, which is characterized in that the signal inspection device be 488nm
During the laser of wavelength, microbial cell and non-microorganism are identified according to the scatter diagram of the scattered light intensity and fluorescence intensity
Cell includes：
The particle of only 488nm scattering light is determined as non-microorganism particle.
4. water quality detection method according to claim 2, which is characterized in that according to the scattered light intensity and fluorescence intensity
Scatter diagram identify microbial cell and non-microorganism particle after, further include：
Micro-organisms living cell and microorganism damaged cell are identified according to the fluorescence intensity of microbial cell.
5. water quality detection method according to claim 4, which is characterized in that the signal inspection device be 488nm
During the laser of wavelength, micro-organisms living cell and microorganism damaged cell bag are identified according to the fluorescence intensity of microbial cell
It includes：
Fluorescence intensity will be inspired more than first threshold, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, but cannot
Fluorescence intensity is inspired more than second threshold, wavelength is 560nm~720nm, and wave crest is determined as micro- life for the particle of 617nm fluorescence
Object living cells；
Fluorescence intensity will be inspired more than the 3rd threshold value, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, while
Fluorescence intensity can be inspired more than the 4th threshold value, wavelength is 560nm~720nm, and wave crest is determined as micro- for the particle of 617nm fluorescence
Biological damaged cell；And/or fluorescence intensity can only be inspired more than the 5th threshold value, wavelength is 560nm~720nm, and wave crest is
The particle of 617nm fluorescence is determined as microorganism damaged cell.
6. water quality detection method according to claim 4, which is characterized in that identified according to the fluorescence intensity of microbial cell
Go out after micro-organisms living cell and microorganism damaged cell, further include：
According to the quantity and micro- life of the scattered light intensity of microbial cell and the scatter diagram of fluorescence intensity detection micro-organisms living cell
Object damaged cell quantity.
7. water quality detection method according to claim 6, which is characterized in that according to the scattered light intensity of microbial cell and
After the scatter diagram detection quantity of micro-organisms living cell and microorganism damaged cell quantity of fluorescence intensity, following formula meters are utilized
Calculate microbial cell concentration：
ρi=ni/(V·w)；
Wherein, ρiRepresent microbial cell concentration, niRepresent the quantity of micro-organisms living cell and microorganism damaged cell, V expressions are treated
The volume of sample water is detected, the volume before the dilution of detected sample water and the detected sample described in w expression dilute samples
The ratio of the volume of water.
8. according to any water quality detection method in claim 1 to 7, which is characterized in that the fluorescent dye includes the
One fluorescent dye and the second fluorescent dye；Wherein, the first fluorescent dye use permeable living cells cell membrane, and in cell
DNA has the reagent of compatibility；Second fluorescent dye, which uses, by incomplete dead cell or to be damaged the thin of cell
After birth, and have with DNA in cell the reagent of compatibility.
9. water quality detection method according to claim 8, which is characterized in that first fluorescent dye is SYBR Green
I；Second fluorescent dye is Presidium Iodide.
10. water quality detection method according to claim 1, which is characterized in that mix detected sample water with fluorescent dye
It closes, before the detected sample water after being dyed, further includes：
Using detected sample water described in dilution reagent dilutions；
Reagent, the detected sample water and the fluorescent dye mixing are diluted by described using oscillator, obtain mixing
Solution.
11. water quality detection method according to claim 10, which is characterized in that the dilution reagent for distilled water and/or
Deionized water.
12. water quality detection method according to claim 10, which is characterized in that the method further includes：
Temperature is used to be protected from light the solution for cultivating the mixing for 28 DEG C to 40 DEG C of constant temperature incubator.
13. water quality detection method according to claim 10, which is characterized in that dilute using distilled water and/or deionized water
Before releasing the detected sample water, further include：
Filter the impurity in the detected sample water.
14. a kind of water quality detection system, which is characterized in that including：
Sample treatment modules (100), for detected sample water to be mixed with fluorescent dye, the detected sample after being dyed
Water；Wherein, the fluorescent dye is the reagent that fluorescence is generated under predetermined wavelength laser excitation；
Detection module (200) swashs the detected sample water after the dyeing for passing through signal inspection device (300)
Light irradiates, and obtains scattered light intensity and/or fluorescence intensity；
Analysis module (400), for being obtained according to the scattered light intensity and/or fluorescence intensity to the detected sample water
Testing result.
15. water quality detection system according to claim 14, which is characterized in that signal inspection device (300) bag
It includes：Sequentially connected laser (301) and exciting light reshaper (302)；Wherein, the exciting light reshaper (302) is used to incite somebody to action
Circular laser facula is shaped as with detecting the comparable oval shaping hot spot of particle size in the detected sample water.
16. according to the water quality detection system described in claim 14, which is characterized in that the detection module (200) further includes：First
Detector (201), the second detector (202), the 3rd detector (203), the 4th detector (204) and flow chamber (205)；Its
In, the detected sample water is housed in the flow chamber (205)；First detector (201) is described to be checked for detecting
The fluorescence that microorganism damaged cell in sample water is sent by shaping hot spot；Second detector (202) is used to detect
The fluorescence that micro-organisms living cell or damaged cell in the detected sample water are sent by shaping hot spot；Described 3rd visits
Device (203) is surveyed for detecting the side scattered light that the biologic grain in the detected sample water is sent by shaping hot spot；Institute
The biologic grain that the 4th detector (204) is stated for detecting in the detected sample water is dissipated by the forward direction that shaping hot spot is sent
Penetrate light.
17. water quality detection system according to claim 16, which is characterized in that first detector (201) with it is described
Include one or more two points of mirrors (206) and one or more band pass filter (207) between shaping hot spot；It is described
Include one or more two points of mirrors (208) and one or more band logical between second detector and the shaping hot spot
Filter plate (209)；Include one or more two points of mirrors (208) between 3rd detector (203) and the shaping hot spot,
And one or more band pass filter (210).
18. water quality detection system according to claim 17, which is characterized in that two points of mirrors (206) are high anti-low pass
Two points of mirrors.
19. water quality detection system according to claim 18, which is characterized in that the anti-low pass two of height divides the back wave of mirror
A length of 560nm~720nm, transmission peak wavelength are less than 560nm；Alternatively, the anti-low pass two of height divide the reflection wavelength of mirror for 490nm~
550nm, transmission peak wavelength are less than 490nm.
20. water quality detection system according to claim 16, which is characterized in that first detector (201) is snowslide
Diode or photomultiplier；Second detector (202) is avalanche diode or photomultiplier；3rd detector
(203) it is avalanche diode or photomultiplier；4th detector (204) is photodiode.
21. water quality detection system according to claim 16, which is characterized in that the analysis module (400) is additionally operable to root
According to first detector (201), second detector (202), the 3rd detector (203) and the 4th detector
(204) scatter diagram for collecting the scattered light intensity and fluorescence intensity identifies microbial cell and non-microorganism particle.
22. water quality detection system according to claim 16, which is characterized in that in the signal inspection device (300)
For 488nm wavelength laser when, the analysis module (200) is additionally operable to will be only in the 3rd detector (203) and institute
It states the 4th detector (204) and detects that the particle during scattering light of 488nm wavelength is determined as non-microorganism particle.
23. water quality detection system according to claim 16, which is characterized in that in the signal inspection device (300)
For 488nm wavelength laser when, the analysis module (400) is additionally operable to：
Fluorescence intensity will be inspired more than first threshold, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, but cannot
Fluorescence intensity is inspired more than second threshold, wavelength is 560nm~720nm, and wave crest is determined as micro- life for the particle of 617nm fluorescence
Object living cells；
Fluorescence intensity will be inspired more than the 3rd threshold value, wavelength is 490nm~550nm, and wave crest is 521nm fluorescence, while
Fluorescence intensity can be inspired more than the 4th threshold value, wavelength is 560nm~720nm, and wave crest is determined as micro- for the particle of 617nm fluorescence
Biological damaged cell；And/or fluorescence intensity can only be inspired more than the 5th threshold value, wavelength is 560nm~720nm, and wave crest is
The particle of 617nm fluorescence is determined as microorganism damaged cell.
24. water quality detection system according to claim 16, which is characterized in that the analysis module (200) is additionally operable to root
It is damaged according to the quantity and microorganism of the scatter diagram detection micro-organisms living cell of the scattered light intensity and fluorescence intensity of microbial cell
Cell quantity.
25. according to any water quality detection system in claim 14 to 24, which is characterized in that the fluorescent dye includes
First fluorescent dye and the second fluorescent dye；Wherein, the first fluorescent dye uses the cell membrane of permeable living cells, and and cell
Middle DNA has the reagent of compatibility；Second fluorescent dye, which uses, by incomplete dead cell or to be damaged cell
Cell membrane, and have with DNA in cell the reagent of compatibility.
26. water quality detection system according to claim 25, which is characterized in that first fluorescent dye is
SYBRGreen I；Second fluorescent dye is Presidium Iodide.
27. water quality detection system according to claim 25, which is characterized in that the sample treatment modules (100) are also wrapped
It includes：First constant displacement pump (101), the second constant displacement pump (102), the 3rd constant displacement pump (103) and the 4th constant displacement pump (104)；
First constant displacement pump (101) extracts the detected sample after the filtering of the first setting capacity from sample bottle (105)
Water is into mixer (106)；Second constant displacement pump (102) extracts the second setting capacity from dilution reagent fluid reservoir (107)
Dilution reagent into the mixer (106)；3rd constant displacement pump (103) is from the first fluorescent dye fluid reservoir (108)
First fluorescent dye of the 3rd setting capacity is extracted into mixer (106)；4th constant displacement pump (104) is glimmering from second
Second fluorescent dye of the 4th setting capacity is extracted in photoinitiator dye fluid reservoir (109) into the mixer (106).
28. water quality detection system according to claim 25, which is characterized in that first constant displacement pump (101), described
Two constant displacement pumps (102), the 3rd constant displacement pump (103) and the 4th constant displacement pump (104) are plunger pump or syringe pump.
29. water quality detection system according to claim 25, which is characterized in that the sample treatment modules further include：The
One solenoid valve (110), second solenoid valve (111), the 5th constant displacement pump (112) and the 6th constant displacement pump (113)；
First solenoid valve (110) is fixed with the described 5th by the pipeline of the sample bottle (105) equipped with the detected sample water
The pipeline of amount pump (112) is connected, and carries out the quantitative extraction to the detected sample water, and the detected sample water of extraction is defeated
It send to mixer (106), alternatively, first solenoid valve (110) will dilute the pipeline and the described 5th of reagent fluid reservoir (107)
The pipeline of constant displacement pump (112) is connected, and carries out the quantitative extraction to diluting reagent, and the dilution reagent of extraction is delivered to described mix
Clutch (106)；
The second solenoid valve (111) is by the pipeline of the first fluorescent dye fluid reservoir (108) and the 6th constant displacement pump (113)
Pipeline is connected, and carries out the quantitative extraction to the first fluorescent dye, and the first fluorescence dye of extraction is delivered to the mixer
(106), alternatively, the second solenoid valve (111) is by the pipeline of the second fluorescent dye fluid reservoir (109) and the 6th constant displacement pump
(113) pipeline is connected, and carries out the quantitative extraction to the second fluorescent dye, and the second fluorescent dye of extraction is delivered to described
Mixer (106).
30. water quality detection system according to claim 29, which is characterized in that the 5th constant displacement pump (112) and described
6th constant displacement pump (113) is plunger pump or syringe pump；First solenoid valve (110) and the second solenoid valve (111) are two
Position-3-way solenoid valve.
31. according to any water quality detection system in claim 27 or 29, which is characterized in that the dilution reagent is steaming
Distilled water and/or deionized water.
32. according to any water quality detection system in claim 27 or 29, which is characterized in that the mixer (106)
Including oscillator, the oscillator be used for by it is described dilution reagent, the detected sample water, first fluorescent dye and
The second fluorescent dye mixing, obtains the solution of mixing.
33. the water quality detection system according to claim 27 or 29, which is characterized in that the mixer (106) is arranged at
Temperature is in 28 DEG C to 40 DEG C of constant temperature incubator (114).
34. water quality detection system according to claim 27, which is characterized in that the flow chamber (205) mixes with described
The first negative pressure sampling pump (115) is provided between device (106), the first negative pressure sampling pump (115) is used for from the mixer
(106) mixed solution is extracted.
35. water quality detection system according to claim 14, which is characterized in that the sample treatment modules (100) are also wrapped
The second negative pressure sampling pump (116) is included, the second negative pressure sampling pump (116) is used to that initial condition to be extracted to obtain test sample to be checked
Product water.
36. water quality detection system according to claim 27, which is characterized in that the sample treatment modules (100) are also wrapped
Include the 3rd solenoid valve (117), the 4th solenoid valve (118), the 3rd negative pressure sampling pump (119) and waste liquid tank (120)；
The 3rd solenoid valve (117) is controlled by the pipeline of the mixer (106) and the 3rd negative pressure sampling pump (119) pipe
Road is connected, and the solution in the mixer (106) is transported to the waste liquid tank using the 3rd negative pressure sampling pump (119)
(120) in；The 3rd solenoid valve (117) is controlled to pump the pipeline of the mixer (106) and the 3rd negative pressure sampling
(119) after pipe cutting, the 4th solenoid valve (118) is controlled to take out pipeline in sample bottle (105) and the 3rd negative pressure
The pipeline of sample pump (119) is connected, and recycles the 3rd negative pressure sampling pump (119) by the filtering sample in the sample bottle (105)
Product are transported in the waste liquid tank (120).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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