CN106574222A - Method for preparing a collecting plate, and an apparatus for measuring floating organisms - Google Patents
Method for preparing a collecting plate, and an apparatus for measuring floating organisms Download PDFInfo
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- CN106574222A CN106574222A CN201580042779.6A CN201580042779A CN106574222A CN 106574222 A CN106574222 A CN 106574222A CN 201580042779 A CN201580042779 A CN 201580042779A CN 106574222 A CN106574222 A CN 106574222A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/12—Apparatus for enzymology or microbiology with sterilisation, filtration or dialysis means
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
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Abstract
The present invention relates to a method for preparing a collecting plate, and an apparatus for measuring floating organisms. A method for preparing a collecting plate, according to an embodiment, comprises the steps of: preparing a filter for collecting or filtering microorganisms; preparing a solvent for a coating portion for coating the filter; mixing a surfactant into the solvent; preparing a coating portion by diluting the solvent with a bioluminescent material; and coating the filter with the prepared coating portion.
Description
Technical field
The present invention relates to a kind of manufacture method of entrapment plate and plankton measurement apparatus.
Background technology
In recent years, with the appearance of bird flu, new type influenza etc., aerial infection is becoming the problem of social concerns, surveys
The problem of plankton (airborne microbial measurement) receives emphasis and treats in amount air, accordingly
Ground, biosensors market also sharp increase.
The method of plankton, there is culture method, staining etc. in existing measurement air.Wherein, culture method is by sample
The biomone swum in gas is trapped in being adapted to the solid or liquid surface of propagation, and trains under appropriate temperature and humidity conditions
After the foster stipulated time, the method that trapping micro organism quantity is obtained in the colony counts occurred from surface；Staining is after dyeing
Using the method for fluorescence microscope.
In recent years, by using ATP (adenosine triphosphate：Adenosine triphosphate) and fluorescein
(luciferin)/luciferase (luciferase) reacts and the ATP biloluminescence methods of luminous principle, will can disappear from ATP
The time needed for series of steps till except process, ATP extractions, measurement luminous quantity foreshortens to 30 minutes or so such that it is able to
Realize sharp work.
However, in order to be suitable for ATP biloluminescence methods, substantially need ATP extractants, swim micro- life when gaseous state is used it for
During thing measuring system, toxic on human body can wait affects.In addition, in order to be applied to automatic system, needing sustainable supply ATP to extract
There are the problem points of somewhat expensive in agent, sustainable supply ATP extractants conventional at present.
Thus, in order to measure plankton, needing persistently to be put into into the luminous material being about to for plankton
Deng a series of manual work, therefore, for exploitation gaseous state in plankton automatic measurement system there is limitation.
The content of the invention
Invent problem to be solved
The present invention is completed to solve problem as above, be its object is to, there is provided a kind of, need not move through for
The luminous a series of manual work of plankton, for promptly measuring the manufacturer of the entrapment plate of plankton
Method and plankton measurement apparatus.
The technical scheme of solve problem
In order to reach above-mentioned or other purposes, according to an aspect of the present invention, there is provided a kind of manufacture method of entrapment plate, its
In, including：The step of preparing for the filter of trapping or filtering microorganism；Prepare the coating for coating above-mentioned filter
The step of solvent in portion；The step of surfactant is mixed in into above-mentioned solvent：Bioluminescence material is diluted in into above-mentioned solvent
And the step of manufacture coating portion；And by manufacture above-mentioned coating portion be coated on filter the step of.
The step of defoamer is added to the coating portion for diluting bioluminescence material can also be included.
In addition, it can be 0.02% that defoamer is sterilized rear concentration.
Furthermore it is also possible to the step of including the above-mentioned coating portion that enzyme stabilizers are mixed in manufacture.
In addition, enzyme stabilizers may include saccharase or Fructose.
In addition, enzyme stabilizers concentration can be 0.1M～0.5M.
Furthermore it is also possible to including, after above-mentioned coating portion to be coated on above-mentioned filter, the step of the above-mentioned filter of drying
Suddenly.
Furthermore it is also possible to including before above-mentioned coating portion is coated, the step of sterilization to above-mentioned filter.
In addition, above-mentioned solvent can be distilled water.
In addition, above-mentioned solvent includes lytic agent, above-mentioned lytic agent may include extractant (Extractant) or can divide
The buffer agent (Lysis-Buffer) of solution.
In addition, each sq in above-mentioned filter can coat above-mentioned coating portion 0.01ml to 0.1ml.
In addition, above-mentioned sterilization can be wet heat sterilization or high steam sterilization (Autoclave).
In addition, in above-mentioned coating portion, the concentration of above-mentioned surfactant can be 0.1%.
It is above-mentioned or in order to reach other purpose, according to a further aspect of the invention, there is provided a kind of plankton measurement dress
Put, it is characterised in that include：Particle distributary division, it includes trapping or filtering the filter of plankton；Biosolubilization
Portion, the plankton that its dissolving is trapped or filtered by above-mentioned particle distributary division, extracts adenosine triphosphate (ATP：
adenosine triphosphate)；Photo detector, it is arranged at the side of above-mentioned particle distributary division, for detection from above-mentioned
The light of the plankton that particle distributary division sends, extracts the concentration or dustiness of above-mentioned plankton；Display part, its use
In the data that display is detected from above-mentioned photo detector；And flowing generating unit, the opposite side of above-mentioned particle distributary division is arranged at,
For making air flow, in the group being made up of surfactant, bioluminescence material, defoamer and enzyme stabilizers one is selected
Coating substance more than individual is in above-mentioned filter.
In above-mentioned surfactant, the concentration of Triton X-100 (TritonX-100) can be 0.1%.
In addition, above-mentioned bioluminescence material may include luciferin and luciferase.
In addition, in above-mentioned enzyme stabilizers, the concentration of saccharase or Fructose can be 0.1M～0.5M.
Invention effect
According to the invention of motion, need not move through other manual work just can be coated on plankton by luminescent substance
On, so as to have the advantages that the measurement process of simplified plankton.
In addition, surfactant is made an addition to into coating portion, so as to be uniformly distributed in filter with luminescent substance can be made
On advantage.
In addition, defoamer is made an addition to into coating portion, so as to can suppress to be produced on the filter by surfactant
The advantage of film.
In addition, enzyme stabilizers are made an addition to into coating portion, so as to when maintaining the continuous illumination of plankton with long-time
Between, and the advantage of the luminous value of plankton can be increased.
Description of the drawings
Fig. 1 is the concept map for representing plankton measurement apparatus according to an embodiment of the invention.
Fig. 2 is the concept map of the various embodiments for representing particle distributary division.
Fig. 3 is the axonometric chart of entrapment plate according to an embodiment of the invention.
Fig. 4 is the profile of entrapment plate according to an embodiment of the invention.
Fig. 5 is the flow chart of trapping board fabrication method according to an embodiment of the invention.
Fig. 6 is the curve chart of the bioluminescence degree according to enzyme stabilizers concentration for representing one embodiment of the invention.
Fig. 7 is only coated the profile of filter amplifying observation on the filter in the case of bioluminescence material.
Fig. 8 is formed filter amplifying observation on the filter in the case of coating portion according to one embodiment of the invention
Profile.
Specific embodiment
Hereinafter, the embodiment of this disclosure is explained in detail with reference to the accompanying drawings.To the same or like of different accompanying drawings
Element marks identical reference, omits the repeat specification to this.Use in the following description for element
Suffix " module " and " portion ", give or mixed only from easily writing from the aspect of description, its own is without mutual
The meaning of difference or effect.In addition, in the embodiment of explanation this disclosure, when being judged as to related known technology
Explanation when not knowing may the main idea of the embodiment of this disclosure, detailed description thereof will be omitted.In addition, accompanying drawing
It is for only for ease of the embodiment for understanding this disclosure, and the technological thought of non-limiting this disclosure, it should be appreciated that
By the present invention include being made in the thought and technical scope of the present invention had altered, equivalent and substitute.
For convenience of description, this specification carries out the luminous plankton measurement apparatus by the use of ATP as one
Explanation.But not limited to this, the entrapment plate of the embodiment of the present invention can be applicable to make the various floating microorganism of microbial luminescence to measure
On device.
Fig. 1 is the concept map for representing plankton measurement apparatus according to an embodiment of the invention, and Fig. 2 is to represent particle
The concept map of the various embodiments of distributary division.
Reference Fig. 1 and Fig. 2, plankton measurement apparatus 1 according to an embodiment of the invention, including：Particle is shunted
Portion 10, it traps microorganism and is used to light；Biosolubilization portion 20, micro- life that its dissolving is trapped by above-mentioned particle distributary division 10
Thing and extract the ATP in microorganism (adenosine triphosphate), DNA, RNA etc.；Photo detector 30, it is arranged at
The side of particle distributary division 10 is stated, the light of the microorganism sent from above-mentioned particle distributary division 10 is detected and is extracted the concentration of microorganism
Or pollution level；Display part 40, the data that its display is detected from above-mentioned photo detector 30；And flowing generating unit 50, its
The opposite side of above-mentioned particle distributary division 10 is arranged at, for making air flow.
In FIG, although above-mentioned particle distributary division 10 is expressed as flat plate shape, this is primarily to show above-mentioned particle
Effect between distributary division 10 and mentioned microorganism dissolving portion 20 and photo detector 30, only conceptually illustrates and belongs to trapping
The element of plate 11, and nonspecific above-mentioned particle distributary division 10 shape, construction, above-mentioned particle distributary division 10 be applicable to
The lower various embodiments for illustrating.
Above-mentioned particle distributary division 10 is such as precipitator (electrostatic pricipitator), inertial impactor
(inertial impactor), cyclone separator (cyclone), centrifuge (centrifuge) etc., by solid capture method
Either liquid trap method can trap dust collect plant or mistake of the entrapment plate of the particle in air either with trapping space
The general name of filter system.
Precipitator is to produce electricity when (-) voltage [or (+) voltage] being put on into discharge electrode by DC high voltage
Corona, now, the moon (-) ion [or positive (+) ion] of generation makes the dust particle in gas powered, by electric power to just
In the collection of the utilization electrostatic principle for being applied in colelctor electrode (entrapment plate) movement of (+) voltage [or (-) voltage] and becomeing trapped in
Dirt device.
Fig. 2A illustrates widest line-plate type (wire to plate used in various precipitator structures
Type), between charging wire (charging wire) and entrapment plate (collecting plate) electric field is formed, and is passed through
When between charging wire and entrapment plate charged particle be captured plate trapping.
Inertial impactor has and is set below in accelerating jet (acceleration nozzle, impaction nozzle)
It is equipped with the knot of shock plate (impaction plate) or collecting pipe (receiving tube) (hereafter referred to collectively as " entrapment plate ")
Structure.
Fig. 2 B illustrate of this inertial impactor, and the air by accelerating jet or ejiction opening (jet) is by catching
Its flow direction of collection plate turns to 90 °, comprising the particle having in aerial particle more than definite quality because inertia flows
Direction not exclusively turns to and impinges upon entrapment plate, the plate that is captured trapping.
Cyclone separator as the utilization for being widely used in the solids separated in fluid or separating drop and gas from
One kind of the segregation apparatuss of mental and physical efforts, with many types and specification, Fig. 2 C illustrate of this cyclone separator.
Air comprising particle is flowed into behind the inside of circular cyclone separator along tangential direction, along cylinder shape inner wall rotation
Then vortex flow is formed, this vortex flow continues to cone (cone) region of cyclone separator bottom, and passes through
Centrifugal force is pushed particle to inwall side and is separated from flowing, removes the flowing (air) of particle from conical lower end portion towards upper
Portion rises and passes through outlet and discharge, by detached particle along conical wall decline by dust to dust hopper (dust
Hopper) etc. (hereafter referred to collectively as " entrapment plate ").
Centrifuge is the device using the lasting centrifugal force occurred during quick rotation, in addition cyclone separator also with
The segregation apparatuss of centrifugal force, centrifuge can will be contained in air using the rotation container of high speed rotation compared with cyclone separator
In particle to rotation container lateral wall separate.
It is adapted to be applied to Large Copacity or high flow capacity because the precipitator pressure loss is low, for nano-scale (100nm
Fine particle below) also has high dust collecting effect.In contrast to this, inertial impactor, cyclone separator etc. are due to letter
Single structure, so having the advantages that cost and maintenance cost are low.
Solid capture method makes sample air be attracted by the particle layer of solid as basis, by absorption, instead
The method for making the material that will be measured be trapped by solid should be waited, can be suitable for plankton air be trapped into above-mentioned tool
The process of standby entrapment plate or trapping space case on above-mentioned particle distributary division 10.
Liquid trap method is sample air is passed through liquid or is contacted with liquid surface, by dissolving, reaction, is sunk
The method to trap the material that will be measured such as shallow lake, suspension, the species of absorbing liquid can be different according to trapping object material.
The liquid for either trapping spatially coating using the entrapment plate in above-mentioned particle distributary division 10 or housing liquid is caught
Diversity method, it is also possible to trap the plankton in air.
Additionally, also applicable by above-mentioned particle distributary division 10, sample air is trapped by filtering material will be surveyed
The trap filter method of the material of amount, sample air is contacted with the pipe etc. of cooling carries out the condensing material for trapping will measure afterwards
The condensing capture method of cooling, sample air is not dissolved, is reacted, is adsorbed and be directly captured bag, trapping bottle, vacuum are caught
The direct capture method of the trappings such as collection bottle, injection tube (syringe), is analyzed after being trapped using molecular diffusion principle
Diffusion capture method.
When the microorganism swum in an atmosphere passes through above-mentioned particle distributary division 10, trapped by above-mentioned particle distributary division 10, shape
Possess on the above-mentioned particle distributary division 10 of microorganism trapping and be coated with catching for ATP reaction luminous agents for ATP bioluminescences
Collection plate 11.
Therefore, when microorganism passes through or become trapped in constitute above-mentioned by above-mentioned entrapment plate, filtering material, filter system
During entrapment plate 11, ATP bioluminescences are formed.
Mentioned microorganism dissolving portion 20 is used as molten using ion, the electromagnetic force of electronics, antibiotic substance, heat energy, catalyst etc.
Solution is trapped or dissolved to the microorganism of the effluent disorder of internal organs of above-mentioned particle distributary division 10 by above-mentioned particle distributary division 10 extracts micro- life
The general designation of the device element of ATP (adenosine triphosphate), DNA, RNA in thing etc., here, dissolving is micro-
What biology referred to is not to melt microorganism and make liquid condition, but a microorganism is decomposed into into multiple key elements or is carried
Take the meaning of multiple key elements.
When being constituted mentioned microorganism dissolving portion 20 with ion generator, the diameter for possessing the discharge tip of ion generator is got over
Big power consumption more rises, and when power consumption rises, not only producing ion can also produce harmful ozone (ozone), therefore, it is excellent
Select ozone free (ozone-free) ion generator of the use carbon brush of a diameter of less than 10 μm of discharge tip.
Using ozone free (ozone-free) ion generator of a diameter of less than 10 μm of discharge tip of carbon brush, have
The low power consumption of below 4W, therefore the ozone for producing is less than 0.01ppm, thus, it is possible to stably meet office air manager
Pin, the ozone management standard of below the 0.06ppm of the 27th article of Section 1 of Industrial Security hygiene.
When being constituted mentioned microorganism dissolving portion 20 with ion generator, by being attached between the charged ion of microorganism
Repulsion damages the cell wall of microorganism and extracts ATP, when being constituted mentioned microorganism dissolving portion 20 with plasma discharge device, by height
The electric discharge of voltage and the ion of high concentration, the impact damage cell wall of electronics that produce and extract ATP.
The ATP extracted by mentioned microorganism dissolving portion 20, be exposed to microorganism cell wall outside while with it is upper
State the reaction luminous agents of the ATP in particle distributary division 10 to be reacted and produced light, by the photodiode for converting light to electricity
(PD), the photo detector 30 such as avalanche photodide (APD) to be detected and extract microorganism by the light that ATP bioluminescences are produced
Concentration or pollution level.
All biologies are stored temporarily in the energy produced by the oxidation in Organic substance in the compound for being called ATP, according to
Need to be hydrolyzed, moved using the energy now released, keep body temperature, this ATP to produce bio electricity sometimes,
Sometimes bioluminescence can also be produced.
Photo detector be measured by the way that the energy for absorbing element is converted to into the form that can be measured photon beam or
(Opticalpower) element of luminous power, has the advantages that high sensitive, fast response time, the minimum noise for starting wavelength,
Therefore, as the element of detection optical signal in the fiberopticscommunication system that near infrared range (0.8～1.6 μm) is operated
Widely use.
Especially, photoelectric cell (photoelectric detectors) is the light by being absorbed in element in photo detector
Son is produced such as electronics (electron), the carrier (carrier) of hole (hole), according to this carrier in element material
The electric current that flowing generation can be measured, i.e. the element based on photoelectricity effect (photo effect), it is adapted to suitable for this
It is bright.
In electromagnetic wave, the eyes for making people feel that the wave-length coverage of bright light is about 380nm to 780nm, as monochromatic light from
Wavelength it is short be followed successively by livid purple 400～500nm, 450～500nm of green grass or young crops, green 500～570nm, 570～590nm of Huang, orange 590～
610nm, 610～700nm of redness, above-mentioned photo detector 30 has can receive 400nm with the wavelength broadband of up to below 700nm
Photo sensitivity.
During plankton in air to be trapped in above-mentioned particle distributary division 10, using such as aerator, the stream of pump
Dynamic generating unit 50 produces the draught head for making the air of the above-mentioned side of particle distributary division 10 to opposite side forced flow.Specifically,
Mentioned microorganism dissolving portion 20 and photo detector 30 are arranged at the upper of the path as Atmospheric Flow to above-mentioned particle distributary division 10
The side of particle distributary division 10 is stated, above-mentioned flowing generating unit 50 is arranged at the opposite side of above-mentioned particle distributary division 10.
The amount of the higher ATP for extracting of the concentration of microorganism is more, and in addition luminosity also can strengthen, and above-mentioned photo detector 30 will
The light of reception be converted to as voltage, electric current, the signal of telecommunication of frequency (frquency) and export.Microorganism concn calculating part (is not schemed
Show) carry out digitization by the signal of telecommunication sent from above-mentioned photo detector 30 and according to the relation of the bioluminescence value of microorganism concn
Or formulation or compare the value of digitization or formulation.
From the light that above-mentioned photo detector 30 is detected, by mentioned microorganism concentration calculating part etc. through formulation or number
According to the signal processing changed, the concentration or pollution level of microorganism are shown in real time by display part 40.
Above-mentioned particle distributary division 10 includes, traps the entrapment plate 11 of microorganism particle.Hereinafter, above-mentioned entrapment plate is described in detail
11 composition.
Fig. 3 is the axonometric chart of entrapment plate according to an embodiment of the invention, and Fig. 4 is side-looking according to an embodiment of the invention
Profile.
Reference Fig. 3 and Fig. 4, entrapment plate according to an embodiment of the invention 11, including：Filter 12, it is floating for trapping
Trip microorganism；Coating portion 14, is coated on above-mentioned filter 12, for making plankton light.
Above-mentioned coating portion 14 is luminescent solution to be coated on above-mentioned filter 12 and is formed.
In this manual, filter 12 be in order to measure plankton for the filter paper that filters, trap, trapping
The general name of substrate.Therefore, above-mentioned filter 12 is formed as the porous filter of air flow or swims for trapping
The variform of the plate shape state of microorganism etc..
Above-mentioned filter 12 is generally formed into plate shape state.Above-mentioned filter 12 is in plankton measurement apparatus described later
As an element for being included in particle distributary division, caught by above-mentioned filter 12 comprising aerial plankton
Collection.For plankton measurement apparatus filter be diameter about 15mm plate, but not limited to this are formed as various sizes
And form.
The one side of above-mentioned filter 12 or two sides possess the coating portion 14 for making plankton luminous.That is, it is interpreted as
Above-mentioned coating portion 14 is, in order to measure the concentration of plankton, to make what the plankton trapped by above-mentioned filter 12 lighted
Constitute.
Hereinafter, the process being coated in coating portion 14 on above-mentioned filter 12 is described in detail.
Fig. 5 is the flow chart of trapping board fabrication method according to embodiments of the present invention.
First, it is possible to be cut into for the disk of above-mentioned filter 12 and can be used in the big of plankton measurement apparatus
Little (S100), in order to the micronic dust and antibacterial of removing remaining are sterilized (S110) to the above-mentioned filter 12 for cutting.To above-mentioned
It is to reduce the error of plankton measurement that filter 12 carries out sterilizing.
Have as sterilization mode, using the wet heat sterilization mode of steam, using the high steam sterilization mode of steam under pressure.
Wet heat sterilization is that high steam sterilization is resistance in order to what is reacted and used in high pressure, high temperature using the warm mode of steam
Pressure vessel or the wet sterilization mode according to hyperpyrexia.
It is however not limited to this, in order to remove micronic dust and the antibacterial remained on above-mentioned filter, using various sterilization
Mode.
Prepare the solvent (S200) for preparing the above-mentioned coating portion 14 of manufacture after above-mentioned filter 12.Above-mentioned coating portion 14 is used
As the lytic agent of solvent.Buffer agent (the Lysis- that above-mentioned lytic agent includes extractant (Extractant), can decompose
) or distilled water Buffer.Above-mentioned solvent passes through the sterilization process same with above-mentioned filter 12.
Complete that surfactant is devoted into above-mentioned lytic agent (S300) after sterilization process.If not putting into above-mentioned surface activity
Agent and only bioluminescence material is devoted into lytic agent, then bioluminescence material can not equably be mixed in lytic agent so as to biology
Luminescent substance is difficult to light.Therefore, surfactant put in above-mentioned lytic agent, so as to bioluminescence in the present embodiment
Material can uniformly be mixed in lytic agent.Used as one, above-mentioned surfactant can use Triton X-100
(TritonX-100)。
Above-mentioned surfactant is put in above-mentioned lytic agent with about 0.1% or so ultimate density.
Surfactant is mixed in after lytic agent and dilutes bioluminescence material (S400).As one, above-mentioned biology
Luminescent substance can be luciferin (Luciferin).Bioluminescence be certain organic compound aoxidized by the effect of enzyme and
The energy of releasing discharges a kind of external photochemical reaction with the form of luminous energy, ties as the luciferin and ATP of luminescent substance
Close and form the complex of luciferin-ATP, and generate the molecules of inorganic phosphate H3PO4 two.Here, luciferin be reduced form so as to
It is labeled as LH2.(LH2+ATPLH2-AMP+2H3PO4)
The LH2-AMP produced in above-mentioned reaction is reacted with oxygen and is in an unsure state, this unstable
The oxidation product of state decomposes and generates oxidized form luciferin and AMP and produce light (hv) at once.Here, L refers to oxidized form
Luciferin, L-AMP* refers to the luciferin-AMP complex (LH2-AMP+1/2O2L-AMP*+ of unstable energy state
H2O) [L-AMP* → L+AMP+hv (luminous energy)].
Bioluminescence material includes luciferase as the enzyme for playing catalytic action.
In the case where the catalytic action of the enzyme of luciferase (luciferase) is called, LH2-AMP is carried out anti-with oxygen (1/2O2)
The process answered and aoxidize, therefore, bioluminescence is that occur in the case where there is luciferin, ATP, luciferase and oxygen, and is calculated
It is that a luciferin molecule releases a photon in oxidizing process.
Therefore, when being constituted bioluminescence material with luciferin (luciferin), by process as above, Neng Gou
Plankton is promptly measured within five minutes, the maximum of light intensity can be measured within three minutes (180sec), therefore,
Within time of measuring can shorten to 3 three minutes.
Above-mentioned bioluminescence material is with the concentration dilution of 400mg/ml in above-mentioned lytic agent.
Bioluminescence material is diluted in after above-mentioned lytic agent and adds defoamer (S500).Above-mentioned defoamer is being killed
In the state of bacterium, can be added with substantially 0.02% concentration.In order to bioluminescence material is equably mixed in into above-mentioned lytic agent
In and add surfactant, therefore, film can be formed on above-mentioned lytic agent surface by the chemical characteristic of above-mentioned surfactant.
Thus, because coating portion 14 can be disproportionately distributed on above-mentioned filter, therefore, can by the above-mentioned defoamer of addition
Prevent from forming film on the surface of above-mentioned lytic agent.
Enzyme stabilizers can be added in above-mentioned coating portion 14.(S510).
Fig. 6 is the curve chart of the bioluminescence degree according to enzyme stabilizers concentration for representing one embodiment of the invention.
Transverse axis represents the luminous persistent period in Fig. 6 curve charts, and the longitudinal axis represents luminous intensity.
With reference to Fig. 6, when adding enzyme stabilizers in above-mentioned coating portion 14, it is able to confirm that bioluminescence reaction continues longer
Time.That is, luminous plankton can for longer periods be measured.In addition, being able to confirm that stable by appropriate enzyme
Agent bioluminescence value is also increasing.
Can be using sucrose (Sucrose), Fructose (Fructose) as above-mentioned enzyme stabilizers, above-mentioned enzyme stabilizers are with 0.1
～0.5 molar concentration (M) is made an addition in above-mentioned coating portion 14.
It is luminous molten by what is prepared for forming coating portion 14 after completing the manufacture of above-mentioned filter 12 and luminescent solution
Drop falls within (S600) on above-mentioned filter 12.Above-mentioned coating portion 14 is with each square centimeter of 0.01～0.1ml of area distributions
Mode be formed on above-mentioned filter 12.After dripping above-mentioned luminescent solution, the above-mentioned of above-mentioned luminescent solution will be coated with and caught
Collection plate 11 is dried under the environment that humidity is less than 10% and makes above-mentioned luminescent solution be deposited on above-mentioned filter 12 (S700)
On.Therefore, when above-mentioned luminescent solution is coated on above-mentioned filter 12, holding time at normal temperatures can longlyer be extended.
Fig. 7 is only coated the profile of filter amplifying observation, Fig. 8 on the filter in the case of bioluminescence material
It is to be formed the profile of filter amplifying observation on the filter in the case of coating portion according to one embodiment of the invention.
With reference to Fig. 7, when bioluminescence material (being luciferin as one) is only coated on filter, due to shape on filter
Increase the pressure loss into the film according to bioluminescence material.Therefore, the luminous of the luminescent substance to plankton is reduced
Efficiency.
With reference to Fig. 8, when the coating portion 14 according to the present embodiment is coated with filter, confirms and lived by above-mentioned surface
Property agent and above-mentioned defoamer ensure that porous.Thus, the pressure loss had not only been reduced but also had improve and plankton had been lighted
The luminous efficiency of material.
According to the invention of motion, need not move through other manual work just can be coated on plankton by luminescent substance
On, therefore, there is simplified plankton measurement process.
In addition, have by the way that surfactant is made an addition to into coating portion making luminescent substance be distributed evenly in filter
Advantage.
In addition, have by the way that defoamer is made an addition to into coating portion to suppress to produce the excellent of film in filter by surfactant
Point.
In addition, when there is long-time to maintain the continuous illumination of plankton by the way that enzyme stabilizers are made an addition to into coating portion
Between, and increase the advantage of the luminous value of plankton.
Claims (17)
1. a kind of manufacture method of entrapment plate, wherein, including：
The step of preparing for the filter of trapping or filtering microorganism；
The step of preparing the solvent of coating portion for coating above-mentioned filter；
The step of surfactant is mixed in into above-mentioned solvent；
The step of bioluminescence material is diluted in into above-mentioned solvent and coating portion is manufactured；And
The step of above-mentioned coating portion of manufacture is coated on into above-mentioned filter.
2. the manufacture method of entrapment plate according to claim 1, wherein, also include：
The step of defoamer is added to the coating portion for diluting above-mentioned bioluminescence material.
3. the manufacture method of entrapment plate according to claim 2, it is characterised in that
Defoamer is sterilized rear concentration for 0.02%.
4. the manufacture method of entrapment plate according to claim 1, wherein, also include：
The step of enzyme stabilizers are mixed in the above-mentioned coating portion of manufacture.
5. the manufacture method of entrapment plate according to claim 4, it is characterised in that
Above-mentioned enzyme stabilizers include saccharase or Fructose.
6. the manufacture method of entrapment plate according to claim 4, it is characterised in that
Above-mentioned enzyme stabilizers concentration is 0.1M～0.5M.
7. the manufacture method of entrapment plate according to claim 1, wherein, also include：
After above-mentioned coating portion to be coated on above-mentioned filter, be dried above-mentioned filter the step of.
8. the manufacture method of entrapment plate according to claim 1, wherein, also include：
Before above-mentioned coating portion is coated, the step of sterilization to above-mentioned filter.
9. the manufacture method of entrapment plate according to claim 1, it is characterised in that
Above-mentioned solvent is distilled water.
10. the manufacture method of entrapment plate according to claim 1, it is characterised in that
Above-mentioned solvent includes lytic agent, and above-mentioned lytic agent includes extractant (Extractant) or the buffer agent that can decompose
(Lysis-Buffer)。
The manufacture method of 11. entrapment plates according to claim 1, it is characterised in that
Above-mentioned coating portion 0.01ml to 0.1ml is coated in each sq of above-mentioned filter.
The manufacture method of 12. entrapment plates according to claim 8, it is characterised in that
Above-mentioned sterilization is wet heat sterilization or high steam sterilization (Autoclave).
The manufacture method of 13. entrapment plates according to claim 1, it is characterised in that
In above-mentioned coating portion, the concentration of above-mentioned surfactant is 0.1%.
14. a kind of plankton measurement apparatus, it is characterised in that include：
Particle distributary division, including the filter of trapping or filtration plankton；
Biosolubilization portion, dissolves the plankton for being trapped or being filtered by above-mentioned particle distributary division, extracts adenosine triphosphate
(ATP：adenosine triphosphate)；
Photo detector, is arranged at the side of above-mentioned particle distributary division, micro- from swimming of sending of above-mentioned particle distributary division for detection
Biological light, extracts the concentration or dustiness of above-mentioned plankton；
Display part, for showing the data detected from above-mentioned photo detector；And
Flowing generating unit, is arranged at the opposite side of above-mentioned particle distributary division, for making air flow,
More than one thing is selected in the group being made up of surfactant, bioluminescence material, defoamer and enzyme stabilizers
Matter is coated on above-mentioned filter.
15. plankton measurement apparatus according to claim 14, it is characterised in that
In above-mentioned surfactant, the concentration of Triton X-100 is 0.1%.
16. plankton measurement apparatus according to claim 14, it is characterised in that
Above-mentioned bioluminescence material includes luciferin and luciferase.
17. plankton measurement apparatus according to claim 14, it is characterised in that
In above-mentioned enzyme stabilizers, the concentration of saccharase or Fructose is 0.1M～0.5M.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2014-0107236 | 2014-08-18 | ||
| KR1020140107236A KR102288766B1 (en) | 2014-08-18 | 2014-08-18 | Filter coted Luminescent substance and Microbial measurement apparatus having the same |
| PCT/KR2015/008618 WO2016028061A1 (en) | 2014-08-18 | 2015-08-18 | Method for preparing a collecting plate, and an apparatus for measuring floating organisms |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106574222A true CN106574222A (en) | 2017-04-19 |
Family
ID=55350947
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201580042779.6A Pending CN106574222A (en) | 2014-08-18 | 2015-08-18 | Method for preparing a collecting plate, and an apparatus for measuring floating organisms |
Country Status (3)
| Country | Link |
|---|---|
| KR (1) | KR102288766B1 (en) |
| CN (1) | CN106574222A (en) |
| WO (1) | WO2016028061A1 (en) |
Cited By (2)
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| CN109182110A (en) * | 2018-10-10 | 2019-01-11 | 张朝明 | A kind of efficient and environment-friendly type plankton detection device and its microorganism detection method |
| CN113176118A (en) * | 2021-04-29 | 2021-07-27 | 江苏大学 | Portable gas fax bacterium real-time acquisition and detection device and method |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110079453B (en) * | 2019-05-07 | 2021-01-15 | 中国科学院电子学研究所 | Automatic sampling detection device and method for near space |
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Also Published As
| Publication number | Publication date |
|---|---|
| WO2016028061A1 (en) | 2016-02-25 |
| KR102288766B1 (en) | 2021-08-12 |
| KR20160021973A (en) | 2016-02-29 |
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