CN106053302A - System for detecting floating algae - Google Patents

System for detecting floating algae Download PDF

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Publication number
CN106053302A
CN106053302A CN201610589182.XA CN201610589182A CN106053302A CN 106053302 A CN106053302 A CN 106053302A CN 201610589182 A CN201610589182 A CN 201610589182A CN 106053302 A CN106053302 A CN 106053302A
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light
inner chamber
laser
sample
reflection
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Inventor
严心涛
王策
马玉婷
吴云良
陈忠详
裴智果
钟金凤
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Suzhou Institute of Biomedical Engineering and Technology of CAS
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Suzhou Institute of Biomedical Engineering and Technology of CAS
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Priority to CN201610589182.XA priority Critical patent/CN106053302A/en
Publication of CN106053302A publication Critical patent/CN106053302A/en
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/01Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N2015/03Electro-optical investigation of a plurality of particles, the analyser being characterised by the optical arrangement
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N2015/1006Investigating individual particles for cytology

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  • Chemical & Material Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

The invention discloses a system for detecting floating algae and belongs to the technical field of detection of miniature floating algae. The system comprises a sample feeding device, a laser device, a light detecting device and an analyzing and calculating module, wherein the laser device is used for emitting laser to irradiate alga cells flowing in the sample feeding device; the light detecting device is used for detecting scattered light and/or fluorescent light generated when the laser irradiates the alga cells; the analyzing and calculating module is used for identifying alga cell types and/or calculating alga cell number according to the detected scattered light and/or fluorescent light. The whole system can automatically detect the alga cell number in a sample, human intervention is not needed, and the system is fast in detection, high in efficiency and high in accurate.

Description

A kind of planktonic algae detecting system
Technical field
The present invention relates to miniature planktonic algae detection field, be specifically related to a kind of planktonic algae detecting system.
Background technology
Miniature planktonic algae is often referred to the particle diameter alga cells less than 20 μm, and they are the primary lifes in aquatic ecosystem Product person, its kind is many, quantity wide, distribution is wide, studies its group, quantity and distributed intelligence to understanding lake, marine ecosystems Kinetics is significant, and the exploitation etc. to lake, marine ecosystems protection, contamination monitoring and resource simultaneously carry For theories integration.
At present, the detection method of planktonic algae is had a lot, such as: Chinese patent CN 203759745 U discloses one The counting assembly of phytoplankton and system, utilize high resolution microscope to analyze phytoplankton and counting;Chinese patent literature A kind of method that CN 101624615 A discloses rapidly screening microalgae germplasm with high grease content, uses fluorescence microscope screening height Oils and fats microalgae cell;Chinese patent literature CN 202421056 U discloses a kind of plankton fast monitored device, uses CCD Imaging device is to plankton instant imaging analysis.
For the miniature planktonic algae that small volume, abundance are bigger, above-mentioned employing high resolution microscope or fluorescence Microscopical cervical arthroplasty method depends on the operation of people, thus detection efficiency is low;And use CCD imaging device that algae is carried out The method analyzed, owing to the data volume of imaging causes more greatly detecting speed relatively slowly, the most per second less than hundreds of cell.
Summary of the invention
The technical problem to be solved in the present invention is to overcome in prior art detection method to alga cells quantity Detection speed problem slower, inefficient.
To this end, the embodiment of the present invention provides a kind of planktonic algae detecting system, including: sample feeding device;Laser instrument, uses In sending laser with the alga cells flowed in irradiating described sample feeding device;Light detection device, is used for detecting described laser It is radiated on described alga cells produced scattered light and/or fluorescence;Analytical calculation module, for filling according to described optical detection Put alga cells kind described in the scattered light and/or fluorescence identifying detected and/or calculate the quantity of described alga cells.
Preferably, described sample feeding device includes: flow chamber, and its inwall constitutes coaxial current stabilization inner chamber, poly-successively Burnt inner chamber, detection inner chamber, described current stabilization inner chamber and described detection inner chamber are cylindricality, and described focusing inner chamber is taper, and described spy The area of the cross section surveying inner chamber is less than the area of the cross section of described current stabilization inner chamber;Sample needle tubing, in described focusing The sample stream including described alga cells is injected in chamber, and the region between outer wall and the flow chamber inwall of sample needle tubing is used for injecting Sheath fluid.
Preferably, the shaft section outline of described sample needle tubing front end is the curve seamlessly transitted.
Preferably, the axial distance of the curve seamlessly transitted described in is 1.4-1.6;Outside described sample needle tubing front end Footpath is 350 μm-450 μm, and the external diameter of rear end is 1.6-2;The internal diameter of described sample needle tubing is 150 μm-350 μm.
Preferably, described flow chamber detection inner chamber cross section be the length of side be the square of 200 μm-400 μm;Or, The cross section of the detection inner chamber of described flow chamber is rectangle, and the bond length of rectangle is 180 μm-200 μm, and long edge lengths is minor face 2 to 3 times of length.
Preferably, described light detection device includes forward scattering photo-detector, is arranged at the dead ahead of described laser optical path, The scattered light of produced forward location it is radiated on described alga cells for receiving described laser.
Preferably, described light detection device also includes: the first fluorescent probe, is arranged at the lateral position of described laser optical path Put, be radiated on described alga cells the fluorescence of produced lateral position for receiving described laser;Described lateral position with The flow direction of alga cells stream is vertical, and vertical with the direction of illumination of described laser;First filter plate, be arranged on described laterally On the fluorescence light path of position, it is located between described flow chamber and described first fluorescent probe, is used for filtering out wavelength and is less than The light of the first predetermined wavelength.
Preferably, described light detection device also includes the first dichroscope, is arranged on the fluorescence light path of described lateral position, Between described flow chamber and described first filter plate, it is less than the light of described first predetermined wavelength for reflection wavelength.
Preferably, described light detection device also includes: the second fluorescent probe, is arranged in the first reflection light light path, uses In receiving described first reflection light;The light that described first reflection light is reflected by described first dichroscope;Second filter plate, is arranged Reflect in light light path described first, between described first dichroscope and described second fluorescent probe, be used for filtering out Light outside second predetermined wavelength range.
Preferably, described light detection device also includes the second dichroscope, is arranged in described first reflection light light path, is positioned at Between described first dichroscope and described second filter plate, for reflection wavelength less than described second predetermined wavelength range minima Light.
Preferably, described light detection device also includes: the 3rd fluorescent probe, is arranged in the second reflection light light path, uses In receiving described second reflection light;The light that described second reflection light is reflected by described second dichroscope;3rd filter plate, is arranged Reflect in light light path described second, between described second dichroscope and described 3rd fluorescent probe, be used for filtering out Light outside 3rd predetermined wavelength range.
Preferably, described forward scattering photo-detector, described first fluorescent probe, described second fluorescent probe and At least one in described 3rd fluorescent probe uses avalanche diode APD.
Preferably, described light detection device also includes Signal-regulated kinase, for by described forward scattering photo-detector, institute State the signal of telecommunication of at least one output in the first fluorescent probe, described second fluorescent probe and described 3rd fluorescent probe Be converted to digital signal.
Preferably, it is additionally provided with illuminator between the laser optical path of described laser instrument and described sample feeding device.
Preferably, the cross section of the laser that described laser instrument is sent is circular light spot;Described laser instrument and described sample At least one pair of transverse focusing cylindrical mirror and longitudinal focusing cylindrical mirror is also included, for by described between the laser optical path of sampling device Circular light spot is shaped as oval hot spot.
Preferably, described system also includes flow monitor, for monitoring the flow velocity of sample and/or sheath fluid.
Preferably, described system also includes flow speed controller, for controlling the flow velocity of sample and/or sheath fluid, in making focusing The Reynolds number of intracavity flowing is less than 2300.
Preferably, described analytical calculation module is additionally operable to the scattered light that detects according to described light detection device and/or glimmering Light analyzes the kind of alga cells.
Preferably, described system also includes display module, for showing the quantity that alga cells is overall and/or the most of the same race The quantity of the alga cells of class.
Technical solution of the present invention, has the advantage that
1. the planktonic algae detecting system that the embodiment of the present invention provides, the laser sent by laser instrument irradiates sample feeding The alga cells of flowing in device, by light detection device exploring laser light be radiated on alga cells produced scattered light and/ Or fluorescence, alga cells kind described in the scattered light arrived according to detection by analytical calculation module and/or fluorescence identifying Class and/or the quantity of calculating alga cells, whole system can detect the quantity of alga cells, nothing in sample automatically when detection Needing human intervention, the speed of detection is fast, efficiency is high, accuracy rate is high.
Accompanying drawing explanation
In order to be illustrated more clearly that the specific embodiment of the invention or technical scheme of the prior art, below will be to specifically In embodiment or description of the prior art, the required accompanying drawing used is briefly described, it should be apparent that, in describing below Accompanying drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not paying creative work Put, it is also possible to obtain other accompanying drawing according to these accompanying drawings.
Figure 1A is the theory diagram of planktonic algae detecting system in the embodiment of the present invention 1;
Figure 1B is the shaft section figure of flow chamber in the embodiment of the present invention 1;
Fig. 1 C is the signal of the alga cells that oval hot spot irradiates the interior intracavity of flow chamber detection in the embodiment of the present invention 1 Figure;
Fig. 1 D is that in the embodiment of the present invention 1, the shaft section of sample needle tubing front end is the analogous diagram during straight angle;
Fig. 1 E is that in the embodiment of the present invention 1, the shaft section of sample needle tubing front end is analogous diagram during cone angle;
Fig. 1 F is the structural representation of sample needle tubing in the embodiment of the present invention 1;
Fig. 1 G is that in the embodiment of the present invention 1, sample needle tubing internal diameter is analogous diagram during 1mm;
Fig. 1 H is that in the embodiment of the present invention 1, sample needle tubing internal diameter is analogous diagram during 200 μm;
Fig. 1 I is that in the embodiment of the present invention 1, the cross section detecting inner chamber position of flow chamber is foursquare schematic diagram;
Fig. 1 J is the schematic diagram that cross section is rectangle at the detection inner chamber position of flow chamber in the embodiment of the present invention 1;
Fig. 1 K is the complete optical path schematic diagram of planktonic algae detecting system in the embodiment of the present invention 1;
Fig. 1 L is the theory diagram of a kind of detailed description of the invention of planktonic algae detecting system in the embodiment of the present invention 1.
Detailed description of the invention
Below in conjunction with accompanying drawing, technical scheme is clearly and completely described, it is clear that described enforcement Example is a part of embodiment of the present invention rather than whole embodiments.Based on the embodiment in the present invention, ordinary skill The every other embodiment that personnel are obtained under not making creative work premise, broadly falls into the scope of protection of the invention.
In describing the invention, it should be noted that term " " center ", " on ", D score, "left", "right", " vertically ", Orientation or the position relationship of the instruction such as " level ", " interior ", " outward " they are based on orientation shown in the drawings or position relationship, merely to Be easy to describe the present invention and simplifying describe rather than instruction or the hint device of indication or element must have specific orientation, With specific azimuth configuration and operation, therefore it is not considered as limiting the invention.Additionally, term " first ", " second ", " the 3rd " is only used for describing purpose, and it is not intended that indicate or hint relative importance.
If additionally, the most non-structure of technical characteristic involved in invention described below difference embodiment Become conflict just can be combined with each other.
Embodiment 1
The present embodiment provides a kind of planktonic algae detecting system, as shown in Figure 1A, including sample feeding device 200, also wraps Include laser instrument 100, light detection device 300 and analytical calculation module 400.
Laser instrument 100 is for sending laser with the alga cells flowed in irradiating sample feeding device 200.
Light detection device 300 is radiated on alga cells produced scattered light and/or fluorescence for exploring laser light.
Analytical calculation module 400 is for algae described in the scattered light detected according to light detection device and/or fluorescence identifying Cell category and/or the quantity of calculating alga cells.
Above-mentioned planktonic algae detecting system, the algae that the laser sent by laser instrument is flowed in irradiating sample feeding device Cell, is radiated on alga cells produced scattered light and/or fluorescence by light detection device exploring laser light, passes through analysis meter Calculate alga cells kind described in the scattered light that arrives according to detection of module and/or fluorescence identifying and/or to calculate algae thin The quantity of born of the same parents, whole system can detect the quantity of alga cells in sample automatically when detection, it is not necessary to human intervention, detection Speed is fast, efficiency is high, accuracy rate is high.
As a kind of preferred implementation of the present embodiment, the cross section of the laser that laser instrument 100 is sent is circular light Speckle.Further, at least one pair of transverse focusing post is also included between the laser optical path of laser instrument 100 and sample feeding device 200 Face mirror and longitudinal focusing cylindrical mirror, for being shaped as oval hot spot by circular light spot.Preferably, the major axis of oval hot spot is 60 μm-120 μm, short axle is 10 μm-20 μm.
Preferably, using the laser instrument of 488nm, the cross section of its emergent light is the circular light spot of diameter 1mm-2mm, according to Laser focusing empirical equation:Wherein, d is laser spot diameter at lens focus, and λ is sharp The wavelength of light device, f is lens Jiao's length, and D is laser emitting optical beam spot diameter.Utilize this empirical equation, generally it is convenient to select Go out suitable transverse focusing cylindrical mirror and longitudinal focusing cylindrical mirror.
As a kind of preferred implementation of the present embodiment, the laser optical path of laser instrument 100 and sample feeding device 200 it Between be additionally provided with illuminator, to shorten the air line distance of laser, so reduce system hardware overall size.
As a kind of preferred implementation of the present embodiment, as shown in Figure 1B, sample feeding device 200 includes flow chamber 210 and sample needle tubing 20.
The inwall of flow chamber 210 constitutes coaxial current stabilization inner chamber 211 successively, focuses on inner chamber 212, detection inner chamber 13.Surely Stream inner chamber 211 and detection inner chamber 213 be cylindricality, and this cylindricality can be cylinder, it is also possible to be that polygon is (such as square for cross section Shape) prismatic, or the cylindricality of other forms.Focusing on inner chamber 212 is taper, and this taper can be cone, or pyramid Shape, or the taper of other forms.The area of the cross section of detection inner chamber 213 is less than the area of the cross section of current stabilization inner chamber 211.
Sample needle tubing 20 includes the sample stream of alga cells to focusing inner chamber 212 for injecting, outside sample needle tubing 20 Region between wall and flow chamber 210 inwall is used for injecting sheath fluid, and in Figure 1B, block arrow represents sheath fluid flow direction, thin arrow table Sample product flow direction.
Hot spot is the laser detection inner chamber according to flow chamber 210 of the ellipse of major axis 60 μm-120 μm, short axle 10 μm-20 μm When 213, it is possible to ensure alga cells be with queue form one by one by hot spot irradiation area rather than in parallel through.Ellipse Circular light spot exposes to detect inner chamber 213, and direction of illumination is vertical with cell flow direction, and the short axle of oval hot spot is positioned at carefully On born of the same parents' flow direction, as shown in Figure 1 C.In Fig. 1 C, block arrow represents that sheath fluid flow direction, thin arrow represent sample flow direction, 230 is the alga cells in sample stream, and 240 is elliptical shaped laser spot.
As a kind of detailed description of the invention of the present embodiment, the shaft section of sample needle tubing 20 front end (i.e. needle tubing shower nozzle) is permissible For the straight angle as shown in figure ip, it is also possible to for cone angle as referring to figure 1e.Being found by FLUENT simulation analysis, sample is from the straight angle The injection of needle tubing shower nozzle time, diffusion is relatively big, is unfavorable for focusing on;Sample, when the needle tubing shower nozzle of cone angle penetrates, spreads less.
As a kind of preferred implementation of the present embodiment, in order to ensure that sample stream can keep stable, design sample pin The shaft section outline of pipe 20 front end is the curve seamlessly transitted, as shown in fig. 1f.The fore-end of sample needle tubing 20 such as Fig. 1 F In dashed rectangle 221 shown in, in square frame, the curve of overstriking is the shaft section outline curve of sample needle tubing front end.Sample Needle tubing front end seamlessly transits without wedge angle, outer surface, makes the diffusion when injection of sample stream less, is conducive to focusing on, it is to avoid sample Product stream forms local turbulent.
Further, when the shaft section of sample needle tubing 20 front end (i.e. needle tubing shower nozzle) is cone angle, such as Fig. 1 G, 1H institute Showing, being analyzed the sample needle tubing of internal diameter 1mm and 200 μm respectively by FLUENT simulation software, result shows, when sample pin When bore is bigger, sample can produce whirlpool on needle tubing shower nozzle top, is unfavorable for focusing on;When sample needle tubing internal diameter is less, sample Can there is bigger diffusion on needle tubing shower nozzle top in product, is unfavorable for equally focusing on.
Accordingly, as a kind of preferred implementation of the present embodiment, in order to enable sample stream to keep stable, do not produce whirlpool Whirlpool, is conducive to focusing on, and the axial distance of the curve that design seamlessly transits is 1.4-1.6, as shown in Fig. 1 F 222.Sample The external diameter of needle tubing front end is 350 μm-450 μm, and as shown in Fig. 1 F 223, the external diameter of rear end is 1.6-2, such as in Fig. 1 F 224 Shown in.The internal diameter of sample needle tubing is 150 μm-350 μm, as shown in Fig. 1 F 225.
Preferably, the material of sample needle tubing uses rustless steel or titanium alloy, the most anti-weak acid and weak base, prevents Long Term Contact Sample conductance causes to get rusty;On the one hand intensity is big, it is possible to prevents deformation, and ensures sample needle tubing position in flow chamber 210 inner chamber Put constant;Additionally, when producing this sample needle tubing, rustless steel and titanium alloy material are more suitable for the processing polished of circular arc.
Preferably, as shown in Figure 1 I, the Internal periphery of cross section at detection inner chamber 213 position of flow chamber 210 and outline For concentrical square, and the square length of side of Internal periphery is 200 μm-400 μm.Or, as shown in figure ij, flow chamber 210 The Internal periphery of cross section at detection inner chamber 213 position and outline be concentrical rectangle, and the minor face of the rectangle of Internal periphery A length of 180 μm-200 μm, long edge lengths is 2 to 3 times of bond length, such as 2 times, 2.2 times, 2.5 times, 2.7 times, 2.95 Times, the multiple of other arbitrary numerals between 3 times etc., or numeral 2 to 3.
Preferably, the material of flow chamber 210 uses quartz glass, and on the one hand light transmission is good, on the other hand linear expansion coefficient Little, during laser prolonged exposure, thermal deformation is little.
Fig. 1 K is the complete optical path figure of system.In figure, 100 is laser instrument, and 301 is illuminator, 302 and 303 be a pair horizontal Focusing cylindrical mirror and longitudinal focusing cylindrical mirror, 213 is the detection inner chamber of flow chamber 210.
As a kind of preferred implementation of the present embodiment, as shown in figure ik, light detection device includes that forward scattering light is visited Survey device 304, be arranged at the dead ahead of laser optical path, be radiated on alga cells produced forward location for receiving laser Scattered light.Forward scattering light is also called small angle scattering light, and its intensity is the most linear with the size of alga cells, according to The power of forward scattering light, can separate alga cells with impurity, debris plume, it is also possible to distinguish the cell mass that diameter is different Kind.
Further, light detection device also includes the first fluorescent probe 305 and the first filter plate 306.
First fluorescent probe 305 is arranged at the lateral position of laser optical path, is used for receiving laser and is radiated at alga cells The fluorescence of upper produced lateral position.This lateral position is vertical with the flow direction of alga cells stream, and with the irradiation of laser Direction is vertical.
First filter plate 306 is arranged on the fluorescence light path of lateral position, is located at flow chamber 210 and the first fluorescence detection Between device 305, for filtering out the wavelength light less than the first predetermined wavelength, the most only allow the wavelength light more than the first predetermined wavelength Pass through.Preferably, this first predetermined wavelength is 650nm, and its wavelength is longer, and energy is more weak, first filters the wavelength of this wave band, subtracts Lack filter plate and dichroiscopic quantity that this band wavelength is passed through, it is possible to reduce the energy loss of this low-yield low light level as far as possible.
Further, light detection device also includes the first dichroscope 307, is arranged on the fluorescence light path of lateral position, 210 and first between filter plate 306, for reflection wavelength less than the light of the first predetermined wavelength.Dichroscope can make to be more than The light of predetermined wavelength passes through, and reflection is less than the light of this predetermined wavelength, i.e. when the first predetermined wavelength is 650nm, the one or two simultaneously Color mirror passes through for the light making wavelength be more than 650nm, and the light that reflection wavelength is less than 650nm.
Yet further, light detection device also includes the second fluorescent probe 308 and the second filter plate 309.
Second fluorescent probe 308 is arranged in the first reflection light light path, for receiving the first reflection light.This first reflection The light that light is reflected by the first dichroscope 307;
Second filter plate 309 is arranged in the first reflection light light path, is positioned at the first dichroscope 307 and the second fluorescent probe Between 308, for filtering out the light outside the second predetermined wavelength range, the light in the second predetermined wavelength range is the most only allowed to pass through. Preferably, the second predetermined wavelength range is 488nm ± 40nm.
Yet further, light detection device also includes the second dichroscope 310, is arranged in the first reflection light light path, is positioned at Between first dichroscope 307 and the second filter plate 309, it is less than the light of the second predetermined wavelength range minima for reflection wavelength. When the second predetermined wavelength range is 488nm ± 40nm, the second dichroscope passes through for the light making wavelength be more than 448nm, and instead The ejected wave length light less than 448nm.
Yet further, light detection device also includes the 3rd fluorescent probe 311 and the 3rd filter plate 312.
3rd fluorescent probe 311 is arranged in the second reflection light light path, for receiving the second reflection light.This second reflection The light that light is reflected by the second dichroscope 310.
3rd filter plate 312 is arranged in the second reflection light light path, is positioned at the second dichroscope 310 and the 3rd fluorescent probe Between 311, for filtering out the light outside the 3rd predetermined wavelength range, the light in the 3rd predetermined wavelength range is the most only allowed to pass through. Preferably, the 3rd predetermined wavelength is 592nm ± 43nm.
It should be added that, the first dichroscope and the second dichroscope employed in above-mentioned preferred implementation are all Allowing wavelength to pass through more than the light of predetermined wavelength, reflection wavelength is less than the light of predetermined wavelength simultaneously.In addition, dichroscope also may be used To be to allow wavelength to pass through less than the light of predetermined wavelength, reflection wavelength is more than the light of predetermined wavelength simultaneously.Above-mentioned first dichroscope The arbitrary one in both dichroscopes can be used, corresponding light path, the filter range of filter plate and glimmering with the second dichroscope Photo-detector does the detailed description of the invention of respective change all within protection scope of the present invention.
In different algal species cell, kind and the content of contained pigment are the most different, and primary pigments as contained by chlorella is chlorophyll a, Pigment contained by diatom also has carotene in addition to chlorophyll a, and contained by hidden algae, pigment also has phycoerythrin, cyanophyceae in addition to chlorophyll a Contained pigment also has phycocyanin in addition to chlorophyll a, phycoerythrin.
Owing to these pigments can excite different fluorescence under specific wavelength laser (such as 488nm), by the inspection of fluorescence intensity Survey can realize the analysis of different algal species kind, therefore, according to above-mentioned first fluorescent probe, the second fluorescent probe, the 3rd Fluorescence in scattered light that fluorescent probe is detected, lateral position, in designated wavelength range, it is possible to tell containing not With the alga cells of pigment, can relatively accurately determine the kind of alga cells in conjunction with the size of alga cells.
Preferably, forward scattering photo-detector the 304, first fluorescent probe the 305, second fluorescent probe 308 and the 3rd At least one in fluorescent probe 311 uses avalanche diode APD.APD volume is little, it is simple to realize the system integration, reduces system The volume of system hardware;Only need the high pressure of about 100V, easy to use;Low price, it is possible to greatly reduce the overall one-tenth of system This.A kind of alternative as APD, it is also possible to use photomultiplier tube PMT.
As shown in figure ik, light detection device also includes being arranged between the first fluorescent probe 305 and the first filter plate 306 The first condenser lens 312, be arranged at the second condenser lens between the second fluorescent probe 308 and the second filter plate 309 313, it is arranged at the tertiary focusing lens 314 between the 3rd fluorescent probe 311 and the 3rd filter plate 312, is arranged at flow chamber 210 and first the 4th condenser lenses 315 between dichroscope 307.
The theory diagram of a kind of detailed description of the invention of planktonic algae detecting system is as can be seen in figure il.
As a kind of preferred implementation of the present embodiment, system also includes Signal-regulated kinase 500, for by forward direction The electricity of at least one output in detector for scattered light, the first fluorescent probe, the second fluorescent probe and the 3rd fluorescent probe Signal is converted to digital signal.Analytical calculation module calculates the quantity of alga cells according to this digital signal.Preferably, signal is adjusted Reason module 500 can also be integrated on light detection device.
As a kind of preferred implementation of the present embodiment, analytical calculation module is for detecting according to light detection device Scattered light and/or fluorescence calculate the quantity of described alga cells, including the quantity of the alga cells calculated in sample, and combine stream The density of the flow rate calculation alga cells of the sample that speed watch-dog is monitored, computing formula isWherein ρ is that algae is thin The density of born of the same parents, n is the quantity of alga cells in sample, and V is the flow of sample.
Additionally, analytical calculation module is additionally operable to scattered light and/or the fluorescence analysis algae detected according to light detection device The kind of cell, according to formulaCalculate the density of various alga cells, wherein ρiAlga cells close for certain kind Degree, niFor the quantity of the alga cells of this kind in sample, V is the flow of sample.This analytical calculation module can be embedded Hardware module, it is also possible to be disposed in computer or mobile terminal the software module run.
As a kind of preferred implementation of the present embodiment, system also includes display module 600, is used for showing alga cells Overall quantity, and/or the quantity of different types of alga cells.This display module can be single display screen, as LED shows Display screen, OLED display screen, electric ink display screen etc., it is also possible to be the display screen of computer or mobile terminal.
In order to make alga cells be formed one by one by the alga cells queue of hot spot irradiation area in detection inner chamber 213, According to principle of hydrodynamics, need to ensure sheath fluid retaining layer stream mode in liquid stream focusing, the i.e. Reynolds number of its flowing Less than 2300.Now, in detection inner chamber 213, the diameter of sample stream is compressed to 10 μm-30 μm by sheath fluid, and alga cells leads to one by one Cross hot spot irradiation area.
Therefore, system also should include flow rate monitors 700, monitors and control the flow velocity of sample stream and/or sheath fluid.This flow velocity Sample flow rate and flow information that monitor controller 700 is monitored are also communicated to analytical calculation module in order to calculate alga cells Density.
Obviously, above-described embodiment is only for clearly demonstrating example, and not restriction to embodiment.Right For those of ordinary skill in the field, can also make on the basis of the above description other multi-form change or Variation.Here without also cannot all of embodiment be given exhaustive.And the obvious change thus extended out or Change among still in the protection domain of the invention.

Claims (12)

1. a planktonic algae detecting system, it is characterised in that including:
Sample feeding device;
Laser instrument, for sending laser with the alga cells flowed in irradiating described sample feeding device;
Light detection device, is used for detecting described laser and is radiated on described alga cells produced scattered light and/or fluorescence;
Analytical calculation module, thin for algae described in the scattered light detected according to described light detection device and/or fluorescence identifying Born of the same parents' kind and/or calculate the quantity of described alga cells.
Planktonic algae detecting system the most according to claim 1, it is characterised in that described sample feeding device includes:
Flow chamber, its inwall constitutes coaxial current stabilization inner chamber successively, focuses on inner chamber, detection inner chamber, described current stabilization inner chamber and institute Stating detection inner chamber is cylindricality, and described focusing inner chamber is taper, and the area of the cross section of described detection inner chamber is less than described current stabilization The area of the cross section of inner chamber;
Sample needle tubing, for injecting the sample stream including described alga cells, the outer wall of sample needle tubing to described focusing inner chamber And the region between flow chamber inwall is used for injecting sheath fluid.
Planktonic algae detecting system the most according to claim 2, it is characterised in that the shaft section of described sample needle tubing front end Outline is the curve seamlessly transitted.
Planktonic algae detecting system the most according to claim 3, it is characterised in that described in curve axial that seamlessly transit Distance is 1.4-1.6;The external diameter of described sample needle tubing front end is 350 μm-450 μm, and the external diameter of rear end is 1.6-2; The internal diameter of described sample needle tubing is 150 μm-350 μm.
Planktonic algae detecting system the most according to claim 2, it is characterised in that the horizontal stroke of the detection inner chamber of described flow chamber Cross section be the length of side be the square of 200 μm-400 μm;Or, the cross section of the detection inner chamber of described flow chamber is rectangle, rectangle Bond length be 180 μm-200 μm, long edge lengths is 2 to 3 times of bond length.
Planktonic algae detecting system the most according to claim 1, it is characterised in that described light detection device includes that forward direction dissipates Penetrate photo-detector, be arranged at the dead ahead of described laser optical path, be used for receiving described laser and be radiated at institute on described alga cells The scattered light of the forward location produced.
7. according to the planktonic algae detecting system described in claim 1 or 6, it is characterised in that described light detection device also includes:
First fluorescent probe, is arranged at the lateral position of described laser optical path, is used for receiving described laser and is radiated at described algae The fluorescence of produced lateral position on class cell;Described lateral position is vertical with the flow direction of alga cells stream, and with institute The direction of illumination stating laser is vertical;
First filter plate, is arranged on the fluorescence light path of described lateral position, is located at described flow chamber and described first fluorescence Between detector, for filtering out the wavelength light less than the first predetermined wavelength.
Planktonic algae detecting system the most according to claim 7, it is characterised in that described light detection device also includes first Dichroscope, is arranged on the fluorescence light path of described lateral position, between described flow chamber and described first filter plate, is used for Reflection wavelength is less than the light of described first predetermined wavelength.
Planktonic algae detecting system the most according to claim 8, it is characterised in that described light detection device also includes:
Second fluorescent probe, is arranged in the first reflection light light path, is used for receiving described first reflection light;Described first reflection The light that light is reflected by described first dichroscope;
Second filter plate, is arranged in described first reflection light light path, is positioned at described first dichroscope and described second fluorescence is visited Survey between device, for filtering out the light outside the second predetermined wavelength range.
Planktonic algae detecting system the most according to claim 9, it is characterised in that described light detection device also includes Two dichroscopes, are arranged in described first reflection light light path, between described first dichroscope and described second filter plate, use The light of described second predetermined wavelength range minima it is less than in reflection wavelength.
11. planktonic algae detecting systems according to claim 10, it is characterised in that described light detection device also includes:
3rd fluorescent probe, is arranged in the second reflection light light path, is used for receiving described second reflection light;Described second reflection The light that light is reflected by described second dichroscope;
3rd filter plate, is arranged in described second reflection light light path, is positioned at described second dichroscope and described 3rd fluorescence is visited Survey between device, for filtering out the light outside the 3rd predetermined wavelength range.
12. according to the planktonic algae detecting system described in claim 6,7,9 or 11, it is characterised in that described forward scattering light At least one in detector, described first fluorescent probe, described second fluorescent probe and described 3rd fluorescent probe Use avalanche diode APD.
CN201610589182.XA 2016-07-25 2016-07-25 System for detecting floating algae Pending CN106053302A (en)

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