CN113604337A - Microorganism aerosol sampling bottle - Google Patents
Microorganism aerosol sampling bottle Download PDFInfo
- Publication number
- CN113604337A CN113604337A CN202011003284.1A CN202011003284A CN113604337A CN 113604337 A CN113604337 A CN 113604337A CN 202011003284 A CN202011003284 A CN 202011003284A CN 113604337 A CN113604337 A CN 113604337A
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- Prior art keywords
- sampling bottle
- air inlet
- filter screen
- bottle body
- sampling
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- 238000005070 sampling Methods 0.000 title claims abstract description 230
- 239000000443 aerosol Substances 0.000 title claims abstract description 62
- 244000005700 microbiome Species 0.000 title claims description 16
- 230000000813 microbial effect Effects 0.000 claims abstract description 30
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 60
- 239000000243 solution Substances 0.000 claims description 47
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 40
- 150000001875 compounds Chemical class 0.000 claims description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 238000004140 cleaning Methods 0.000 claims description 22
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 20
- 239000008367 deionised water Substances 0.000 claims description 20
- 229910021641 deionized water Inorganic materials 0.000 claims description 20
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 20
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 18
- 230000000845 anti-microbial effect Effects 0.000 claims description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- FSSPGSAQUIYDCN-UHFFFAOYSA-N 1,3-Propane sultone Chemical compound O=S1(=O)CCCO1 FSSPGSAQUIYDCN-UHFFFAOYSA-N 0.000 claims description 15
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- 238000005406 washing Methods 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
- 229910052757 nitrogen Inorganic materials 0.000 claims description 11
- 239000007789 gas Substances 0.000 claims description 10
- CRVGTESFCCXCTH-UHFFFAOYSA-N methyl diethanolamine Chemical compound OCCN(C)CCO CRVGTESFCCXCTH-UHFFFAOYSA-N 0.000 claims description 10
- 150000008053 sultones Chemical class 0.000 claims description 10
- 150000003512 tertiary amines Chemical class 0.000 claims description 10
- DVKJHBMWWAPEIU-UHFFFAOYSA-N toluene 2,4-diisocyanate Chemical compound CC1=CC=C(N=C=O)C=C1N=C=O DVKJHBMWWAPEIU-UHFFFAOYSA-N 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 8
- 238000006243 chemical reaction Methods 0.000 claims description 6
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
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- 230000001376 precipitating effect Effects 0.000 claims description 5
- 239000011521 glass Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 abstract description 25
- 239000002245 particle Substances 0.000 description 34
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- 241001112090 Pseudovirus Species 0.000 description 2
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- 238000002955 isolation Methods 0.000 description 2
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- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
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- 241001331845 Equus asinus x caballus Species 0.000 description 1
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- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
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- 230000003385 bacteriostatic effect Effects 0.000 description 1
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- 238000010586 diagram Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000012948 isocyanate Substances 0.000 description 1
- 150000002513 isocyanates Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
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- 210000003437 trachea Anatomy 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
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- 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
-
- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
- C03C17/34—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions
- C03C17/3405—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions with at least two coatings of organic materials
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- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/20—Material Coatings
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12M23/46—Means for fastening
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- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
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- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/22—Settling tanks; Sedimentation by gravity
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Abstract
The invention discloses a microbial aerosol sampling bottle, which comprises a sampling bottle body and a sampling bottle cap, wherein the sampling bottle body comprises an air inlet part, a collecting part and a placing part; be equipped with the air inlet on the lateral wall of air inlet portion, be equipped with the filter screen in the sampling bottle, the filter screen passes through the filter screen retainer plate to be fixed at the top of air inlet portion, and the filter screen retainer plate includes fixed part and the outlet duct that is located the fixed part below, is equipped with the connecting hole with the outlet duct intercommunication on the fixed part, and the outlet duct bottom is connected with the filter screen. The cyclone air inlet duct is completely positioned in the sampling bottle, so that the pollution of residual liquid of air inlet equipment to sampling liquid is avoided; and set up the cyclone filter screen in the sampling bottle, wet wall area when having increased the cyclone has effectively reduced the escape of secondary aerosol, has improved sampling efficiency.
Description
Technical Field
The invention relates to the technical field of environment monitoring devices, in particular to a microbial aerosol sampling bottle.
Background
An atmospheric aerosol refers to a stable mixed system of solid fine particles and liquid fine particles uniformly dispersed in the atmosphere, and the fine particles are collectively referred to as aerosol particles. Wherein, the microorganism material or particles with life activity are collectively called microorganism aerosol particles, including microorganisms such as virus, bacteria, fungi, pollen, spore, etc. The aerodynamic diameter of the atmospheric aerosol particles is mostly between 0.001 to 100 μm, and the particles are light enough to suspend in the air. Particles with the particle size of 5-10 mu m can enter human trachea and bronchus, and particles with the particle size of less than 5 mu m can penetrate deep into a deep respiratory tract system. When the concentration of the aerosol reaches a high enough level, it will pose a threat to human health, and especially the microbial aerosol in the air can cause acute and chronic diseases in human beings. Meanwhile, microorganisms can generate various dormant bodies, can survive in the air for a long time, and can be diffused and transmitted by virtue of an air medium, so that outbreak and spread of various infectious diseases are caused, and serious harm is caused. Therefore, how to effectively collect the microbial aerosol in the surrounding environment and monitor the concentration has become a problem to be solved by those skilled in the relevant field.
The wet-wall cyclone method is one of the commonly used microorganism aerosol collection methods at present, and is a method for separating gas from microorganism particles by using the inertia of airflow in a sampling device during high-speed rotation, and absorbing and trapping the microorganism particles through a liquid film formed on the cyclone wall of a sampling bottle. The existing sampling device for the wet-wall cyclone method generally comprises an air inlet device and a sampling bottle connected with the air inlet device. For example, the publication of "a portable wet-wall cyclone microorganism aerosol collector" in the chinese patent document, whose publication number CN111500427A, mainly includes an air inlet isolation hood, an air inlet component, an air duct, a liquid supplementing component, a fan component, a sampling cup, a filter component, a connecting plate, etc., wherein the side surface of the air inlet component is uniformly provided with sharp air inlets, the air inlet isolation hood enters the air inlet component, an air vortex is formed in the air inlet component and the sampling cup through the sharp cut on the air inlet component, the aerosol in the air makes a rotational motion, and the particles and the water mist move to the inner wall of the sampling cup under the action of inertial centrifugal force and are combined with the sampling liquid.
However, in the existing wet-wall cyclone type aerosol collecting device, the air inlet is generally located on the air inlet equipment, so that a part of the cyclone wall is generally located in the air inlet equipment, and a part of the cyclone wall is located in the sampling bottle, and the gas path residue on the cyclone wall in the air inlet equipment is easy to cause cross contamination between sampling at each time, so that the pipeline needs to be replaced to sterilize after sampling at each time, which is very inconvenient. The existing wet wall cyclone type aerosol collecting device has low sampling efficiency, the sampling effect cannot be achieved when the cyclone speed is low, and the cyclone speed is increased, so that a large amount of secondary aerosol is rebounded and is discharged along with the airflow, the sampling effect is greatly reduced, and the application of a wet wall cyclone method is limited.
Disclosure of Invention
The invention aims to overcome the defects that in the existing wet-wall cyclone type microbial aerosol collection device, gas path residues on the cyclone wall in air inlet equipment are easy to cause cross contamination and influence the sampling result; the existing wet-wall cyclone type aerosol collecting device has low sampling efficiency, cannot achieve the sampling effect when the cyclone speed is low, and can cause secondary aerosol to escape when the cyclone speed is increased, so that the sampling effect is greatly reduced, and the application problem of a wet-wall cyclone method is limited; and set up the cyclone filter screen in the sampling bottle, wet wall area when having increased the cyclone has effectively reduced the escape of secondary aerosol, has improved sampling efficiency.
In order to achieve the purpose, the invention adopts the following technical scheme:
a microbial aerosol sampling bottle comprises a sampling bottle body and a sampling bottle cap arranged at the top of the sampling bottle body, wherein the sampling bottle body sequentially comprises an air inlet part, a collecting part communicated with the air inlet part and a placing part which is positioned at the bottom of the collecting part and used for stably placing the sampling bottle from top to bottom; be equipped with the air inlet on the lateral wall of portion of admitting air, be equipped with the toper filter screen in the sampling bottle, the toper filter screen passes through the filter screen retainer plate and fixes the top at the portion of admitting air, the filter screen retainer plate is including being used for with the fixed part of portion of admitting air top fixed connection and being located the outlet duct of fixed part below, be equipped with the connecting hole with the outlet duct intercommunication on the fixed part, the top fixed connection of outlet duct bottom and toper filter screen.
According to the invention, the air inlet part provided with the air inlet is arranged on the sampling bottle body, so that during sampling, air directly enters the sampling bottle body from the air inlet on the sampling bottle body and does not enter the sampling bottle body through the air inlet equipment, and the cyclone wall is completely positioned in the sampling bottle body, thereby reducing sample cross contamination caused by air path residues on the cyclone wall in the air inlet equipment.
Meanwhile, the conical filter screen is arranged in the sampling bottle, so that a second cyclone wet wall (the first is the inner wall of the sampling bottle body, and the common product only has the wet wall) can be formed on the surface of the conical filter screen by using cyclone energy during sampling, the area of the wet wall is obviously increased, and the absorption and collection rate of aerosol particles is improved; and the secondary aerosol in the cyclone cavity is recovered to the utmost extent through the impact between the microbial aerosol particles and the filter screen when the microbial aerosol particles rotate, so that the escape loss of the secondary aerosol is effectively reduced, and the sampling efficiency is obviously improved.
When the sampling bottle is used, the sampling bottle cap is rotated, the sampling liquid is added into the sampling bottle body through the connecting hole and the air outlet pipe of the filter screen fixing ring, then the sampling bottle body is connected with the air inlet device through the connecting hole on the filter screen fixing ring fixing part, after the air inlet device is opened, the sampling liquid forms a sampling liquid film on the inner wall of the sampling bottle and the conical filter screen, air enters the air inlet part of the sampling bottle from the air inlet of the air inlet part, air flow spirally moves from top to bottom along the inside of the sampling bottle, the rotating radius of the air flow is smaller and smaller, and after the vortex reaches the bottom of the sampling bottle body collecting part, the air flow rotates and upwards rotates along the axis, and finally the air flow is discharged through the air outlet pipe on the filter screen fixing ring; meanwhile, the microbial aerosol particles in the air move towards the inner wall of the sampling bottle body under the action of centrifugal force, and under the action of centrifugal force and diffusion, the aerosol particles are absorbed by a collecting liquid film formed by collecting liquid on the inner wall of the sampling bottle body and a filter screen, flow downwards along the inner wall of the bottle body under the combined action of subsequent fluid and gravity, enter a collecting part and are collected, so that the collection of the microbial aerosol is realized; after sampling, the sampling bottle cap is covered, the air inlet is sealed, and the collected sample can be effectively stored.
Preferably, the bottom of the collecting part is in an inverted cone shape, the top of the collecting part is in a cylinder shape, and the conical part of the collecting part has the same taper as that of the conical filter screen. The collection portion top of sampling bottle is cylindrically, can increase the length of cyclone wall, improves the separation effect of aerosol granule and gas, and the collection portion bottom is conical, can play the guide effect to the microorganism aerosol granule of entrapment, makes the downward landing of edge collection portion inner wall that the aerosol granule can be better, gets into the collection portion bottom and collects, has reduced the escape loss of secondary aerosol.
Preferably, the included angle between the inclined sideline of the conical filter screen and the central line is 10-15 degrees, and the mesh number of the conical filter screen is 40-100 meshes. The taper of the conical filter screen is in the range, and is matched with the movement route of the cyclone air path, so that the formation of the cyclone air path is not hindered, and a second cyclone wet wall can be formed on the surface of the conical filter screen by effectively utilizing the cyclone energy; the mesh number of the conical filter screen is in the range, so that gas can effectively pass through, the collision probability of microbial aerosol particles and the conical filter screen is increased, the escape loss of secondary aerosol is effectively reduced, and the sampling efficiency is improved.
Preferably, the inner diameter of the cylindrical parts of the air inlet part and the collecting part is 34-44 mm; the height of the cylindrical part of the air inlet part and the cylindrical part of the collecting part are respectively 20-30 mm and 33-42 mm. The sizes of the air inlet part and the collecting part are in the range, so that the cyclone gas circuit has an optimal motion route, microbial aerosol particles and gas can be effectively separated conveniently, and the sampling efficiency is improved.
Preferably, the ratio of the top surface diameter of the conical filter screen to the inner diameter of the air inlet part is 1: (1.6-2.1). Within the proportion range, the escape loss of the secondary aerosol can be reduced to the maximum extent.
Preferably, the area of the air inlet on the air inlet part is 162-220 mm2. The area of the air inlet can ensure the optimal air inflow in the range, so that an optimal cyclone path can be formed, microbial aerosol particles are effectively separated from air, and the sampling efficiency is improved.
Preferably, the size of the fixing part of the filter screen fixing ring is matched with the inner diameter of the air inlet part of the sampling bottle body, and the fixing part of the filter screen fixing ring is clamped in the air inlet part.
Preferably, the outer wall of the cylindrical part of the collecting part is provided with a positioning rib arranged along the height direction of the sampling bottle body, and the positioning rib is provided with a first positioning groove. The outer wall of the collecting part is provided with a positioning rib arranged along the height direction of the sampling bottle, and the sampling bottle body can be limited in the circumferential direction through the matching of the positioning rib and a positioning device on the air inlet equipment, so that the rotation of the sampling bottle body is avoided when the sampling bottle cover is sampled and rotated; set up first positioning groove on the location muscle, through first positioning groove and the last positioner's of air inlet equipment cooperation, can make and carry on spacingly to the sampling bottle in vertical direction, the up-and-down motion of sampling bottle when avoiding sampling has improved the security and the stability of sampling.
Preferably, the placing part is provided with a second positioning groove. Set up second positioning groove on the portion of placing of sampling bottle, through second positioning groove and the positioner's on the face of placing cooperation, can effectively fix the portion of placing on the face of placing, avoid the sampling bottle in the activity of the face of placing, improved the stability of placing of sampling bottle, avoid the damage of sampling bottle in the sampling process.
Preferably, the sampling bottle body is made of glass, and the sampling bottle body is subjected to antimicrobial adhesion treatment, wherein the treatment method comprises the following steps:
(A) the molar ratio is (2-2.5): 1, mixing toluene diisocyanate and N-methyldiethanolamine, adding the mixture into N, N-dimethylformamide, wherein the mass ratio of the N, N-dimethylformamide to the toluene diisocyanate is (3-4): 1, stirring and reacting at 60-80 ℃ for 12-24 hours under the protection of nitrogen to obtain a tertiary amine intermediate solution;
(B) dissolving 1, 3-propane sultone in N, N-dimethylformamide to obtain a sultone solution, wherein the mass ratio of the 1, 3-propane sultone to the N, N-dimethylformamide is 1: (2-3); dropwise adding a sultone solution into a tertiary amine intermediate solution to enable the molar ratio of 1, 3-propane sultone to N-methyldiethanolamine to be (1-2): 1, stirring and reacting for 20-30 h at 80-100 ℃ under the protection of nitrogen;
(C) adding tetrahydrofuran into the reacted solution, wherein the mass ratio of the reacted solution to the tetrahydrofuran is (4-6): 1, precipitating for 2-4 h, filtering, washing a product with deionized water, and drying in vacuum at 50-60 ℃ for 20-30 h to obtain an isocyanate-terminated zwitterionic compound;
(D) ultrasonically cleaning a sampling bottle for 10-15 min by using a mixed solution of absolute ethyl alcohol and deionized water with the volume ratio of 1 (1-2), washing by using deionized water, and then using N2Drying, then immersing the cleaned sampling bottle body into a toluene solution of 10-15% by mass of 3-aminopropyltriethoxysilane, reacting for 2-4 h at 50-60 ℃, cleaning with absolute ethanol and deionized water, and cleaning with N2Drying to obtain a sampling bottle body with an aminated surface;
(E) dissolving an isocyanate-terminated zwitterionic compound in tetrahydrofuran, adding N, N-diisopropylethylamine, and uniformly stirring to obtain a zwitterionic compound solution, wherein the mass fraction of the isocyanate-terminated zwitterionic compound is 8-12%, and the N, N-diisopropylethylamineThe mass fraction of (A) is 0.3-0.5%; immersing the sampling bottle body with the aminated surface into a zwitterionic compound solution for reaction for 1-3 h, taking out, cleaning with tetrahydrofuran, and using N2And (3) after drying, carrying out heat treatment at 100-120 ℃ for 1-2 h, and cooling to obtain the sampling bottle body subjected to antimicrobial adhesion treatment.
The method comprises the steps of (A) to (C) preparing an isocyanate-terminated zwitterionic compound, modifying amino on the surface of a sampling bottle in the step (D), placing the sampling bottle with the surface aminated in a solution of the isocyanate-terminated zwitterionic compound in the step (E) for reaction, bonding the zwitterionic compound on the surface of the sampling bottle through the addition reaction of the isocyanate and the amino, and finally spontaneously generating a layer of uniformly distributed zwitterionic self-assembled monomolecular film on the surface of the sampling bottle through the self-assembly effect.
In an environment with water, the high-hydrophilicity zwitter-ion macromolecular chain grafted on the surface of the sampling bottle body can strongly interact with free water on the surface through the solvation effect and hydrogen bond interaction of ions to form a hydration layer, so that on one hand, the collection liquid is favorable for forming a wet wall on the inner wall of the sampling bottle body, the collection liquid is promoted to absorb and collect the microbial aerosol particles, and the sampling efficiency is improved; on the other hand, the formation of the hydration layer can establish a barrier on the inner wall of the sampling bottle body, so that the adhesion of microorganism particles on the sampling bottle body is effectively prevented, and the influence of particle adhesion on a detection result and the cross contamination on subsequent sampling are avoided.
Therefore, the invention has the following beneficial effects:
(1) the sampling bottle body is provided with the air inlet part provided with the air inlet, and during sampling, air directly enters the sampling bottle body from the air inlet on the sampling bottle body and does not enter the sampling bottle body through the air inlet equipment, so that the cyclone wall is completely positioned in the sampling bottle body, and the sample cross contamination caused by air path residues on the cyclone wall in the air inlet equipment is reduced;
(2) the filter screen is arranged in the sampling bottle, a second cyclone wet wall can be formed on the surface of the filter screen by cyclone energy during sampling, the area of the wet wall is increased, the acquisition rate of aerosol particles is improved, secondary aerosol in a cyclone cavity is recovered to the maximum extent by the impact between microbial aerosol particles and the filter screen during rotation, the escape loss of the secondary aerosol is effectively reduced, and the sampling efficiency is obviously improved;
(3) the sampling bottle body is subjected to antimicrobial adhesion treatment, and the surface of the sampling bottle body is modified with a high-hydrophilicity zwitterionic compound film, so that on one hand, the collection liquid is favorable for forming a wet wall on the inner wall of the sampling bottle body, the collection liquid is promoted to absorb and collect the microbial aerosol particles, and the sampling efficiency is improved; on the other hand, the formation of the hydration layer can establish a barrier on the inner wall of the sampling bottle body, so that the adhesion of microorganism particles on the sampling bottle body is effectively prevented, and the influence of particle adhesion on a detection result and the cross contamination on subsequent sampling are avoided.
Drawings
Fig. 1 is a schematic view of a sampling bottle cap according to the present invention.
Fig. 2 is a schematic structural diagram of the sampling bottle cap opening device.
Fig. 3 is a schematic illustration of an exploded structure of the present invention.
In the figure: 1 sampling bottle, 101 air inlet portion, 102 collection portion, 103 placing portion, 104 threaded connection portion, 105 air inlets, 106 location muscle, 107 first positioning groove, 108 second positioning groove, 2 sampling bottle lids, 3 filter screen retainer plates, 301 fixed part, 302 outlet duct, 303 connecting hole, 4 toper filter screens.
Detailed Description
The invention is further described with reference to the following detailed description and accompanying drawings.
Referring to fig. 1, a sampling bottle for microbial aerosol used in various embodiments of the present invention includes a glass sampling bottle body 1 and a sampling bottle cap 2 disposed on the top of the sampling bottle body. As shown in fig. 2 and 3, the sampling bottle body sequentially comprises an air inlet part 101, a collecting part 102 communicated with the air inlet part, and a placing part 103 located at the bottom of the collecting part and used for stably placing the sampling bottle, a threaded connection part 104 used for being connected with a sampling bottle cap is arranged on the outer side of the bottle body between the air inlet part and the collecting part, and the sampling bottle cap is sleeved on the outer side of the air inlet part and is in threaded connection with the threaded connection part.
An air inlet 105 is formed in the side wall of the air inlet part, the bottom of the collecting part is in an inverted conical shape, the top of the collecting part is in a cylindrical shape, the conical part has the same taper as the conical filter screen, a positioning rib 106 arranged along the height direction of the sampling bottle body is arranged on the outer wall of the cylindrical part, and a first positioning groove 107 is formed in the positioning rib; the placing part is circular, the center of the placing part is connected with the conical vertex of the collecting part, and the placing part is provided with a second positioning groove 108.
A conical filter screen 4 is arranged in the sampling bottle body, the conical filter screen is fixed at the top of the air inlet part through a filter screen fixing ring 3, and the conical filter screen is coaxial with the conical part of the sampling bottle body collecting part; the filter screen retainer plate is including being used for with inlet portion top fixed connection's fixed part 301 and being located the outlet duct 302 of fixed part below, the top joint of outlet duct bottom and toper filter screen, be equipped with on the fixed part with the connecting hole 303 of outlet duct intercommunication, the fixed part size matches with the inlet portion internal diameter of sampling bottle, the fixed part screens is in the inlet portion of sampling bottle.
When the sampling bottle is used, the sampling bottle cap is rotated, the sampling liquid is added into the sampling bottle body through the connecting hole and the air outlet pipe of the filter screen fixing ring, then the sampling bottle body is connected with the air inlet device through the connecting hole on the filter screen fixing ring fixing part, after the air inlet device is opened, the sampling liquid forms a sampling liquid film on the inner wall of the sampling bottle and the conical filter screen, air enters the air inlet part of the sampling bottle from the air inlet of the air inlet part, air flow spirally moves from top to bottom along the inside of the sampling bottle, the rotating radius of the air flow is smaller and smaller, and after the vortex reaches the bottom of the sampling bottle body collecting part, the air flow rotates and upwards rotates along the axis, and finally the air flow is discharged through the air outlet pipe on the filter screen fixing ring; meanwhile, the microbial aerosol particles in the air move towards the inner wall of the sampling bottle body under the action of centrifugal force, and under the action of centrifugal force and diffusion, the aerosol particles are absorbed by a collecting liquid film formed by collecting liquid on the inner wall of the sampling bottle body and the conical filter screen, flow downwards along the inner wall of the bottle body under the combined action of subsequent fluid and gravity, enter the collecting part and are collected, so that the collection of the microbial aerosol is realized; after sampling, the sampling bottle cap is covered, the air inlet is sealed, and the collected sample can be effectively stored.
Example 1:
the mesh number of the conical filter screen is 100 meshes, the diameter of the top surface is 26mm, and the included angle between the inclined sideline and the central line is 15 degrees; the inner diameter of the cylindrical parts of the air inlet part and the collecting part of the sampling bottle body is 44 mm; the heights of the cylindrical parts of the air inlet part and the collecting part are respectively 30mm and 42 mm; the area of the air inlet on the air inlet part is 220mm2。
Example 2:
the mesh number of the conical filter screen is 60 meshes, the diameter of the top surface is 21mm, and the included angle between the inclined sideline and the central line is 12 degrees; the inner diameter of the cylindrical parts of the air inlet part and the collecting part of the sampling bottle body is 39 mm; the heights of the cylindrical parts of the air inlet part and the collecting part are respectively 25mm and 38 mm; the area of the air inlet on the air inlet part is 180mm2。
Example 3:
the mesh number of the conical filter screen is 40 meshes, the diameter of the top surface is 16mm, and the included angle between the inclined sideline and the central line is 10 degrees; the inner diameter of the cylindrical parts of the air inlet part and the collecting part of the sampling bottle body is 34 mm; the heights of the cylindrical parts of the air inlet part and the collecting part are respectively 20mm and 33 mm; the area of the air inlet on the air inlet part is 162mm2。
Example 4:
the sampling vial of example 4 was subjected to an antimicrobial adherence treatment prior to sampling, the treatment comprising the steps of:
(A) mixing the components in a molar ratio of 2.3: 1, mixing toluene diisocyanate and N-methyldiethanolamine, adding the mixture into N, N-dimethylformamide, wherein the mass ratio of the N, N-dimethylformamide to the toluene diisocyanate is 3.5: 1, stirring and reacting at 70 ℃ for 18h under the protection of nitrogen to obtain a tertiary amine intermediate solution;
(B) dissolving 1, 3-propane sultone in N, N-dimethylformamide to obtain a sultone solution, wherein the mass ratio of the 1, 3-propane sultone to the N, N-dimethylformamide is 1: 2.5; dropwise adding the sultone solution into the tertiary amine intermediate solution to ensure that the molar ratio of the 1, 3-propane sultone to the N-methyldiethanolamine is 1.5: 1, stirring and reacting at 90 ℃ for 24 hours under the protection of nitrogen;
(C) adding tetrahydrofuran into the reacted solution, wherein the mass ratio of the reacted solution to the tetrahydrofuran is 5: 1, precipitating for 3h, filtering, washing a product with deionized water, and drying in vacuum at 55 ℃ for 24h to obtain an isocyanate-terminated zwitterionic compound;
(D) ultrasonically cleaning a sampling bottle for 12min by using a mixed solution of absolute ethyl alcohol and deionized water with the volume ratio of 1:1.5, washing by using deionized water, and then using N2Drying, then immersing the cleaned sampling bottle body into a toluene solution of 3-aminopropyltriethoxysilane with the mass fraction of 12%, reacting for 3h at 55 ℃, cleaning with absolute ethyl alcohol and deionized water, and cleaning with N2Drying to obtain a sampling bottle body with an aminated surface;
(E) dissolving an isocyanate-terminated zwitterionic compound in tetrahydrofuran, adding N, N-diisopropylethylamine, and uniformly stirring to obtain a zwitterionic compound solution, wherein the mass fraction of the isocyanate-terminated zwitterionic compound is 10%, and the mass fraction of the N, N-diisopropylethylamine is 0.4%; immersing the sampling bottle body with aminated surface into the zwitterionic compound solution for reaction for 2h, taking out, cleaning with tetrahydrofuran, and washing with N2After drying, the sample was heat treated at 110 ℃ for 1.5h and cooled to obtain the antimicrobial-treated sample vial.
The rest is the same as in example 1.
Example 5:
the sampling vial of example 5 was subjected to an antimicrobial adherence treatment prior to sampling, the treatment comprising the steps of:
(A) mixing the components in a molar ratio of 2: 1, mixing toluene diisocyanate and N-methyldiethanolamine, and adding the mixture into N, N-dimethylformamide, wherein the mass ratio of the N, N-dimethylformamide to the toluene diisocyanate is 3: 1, stirring and reacting at 60 ℃ for 24 hours under the protection of nitrogen to obtain a tertiary amine intermediate solution;
(B) dissolving 1, 3-propane sultone in N, N-dimethylformamide to obtain a sultone solution, wherein the mass ratio of the 1, 3-propane sultone to the N, N-dimethylformamide is 1: 2; dropwise adding the sultone solution into the tertiary amine intermediate solution to ensure that the molar ratio of the 1, 3-propane sultone to the N-methyldiethanolamine is 1:1, stirring and reacting for 30 hours at 80 ℃ under the protection of nitrogen;
(C) adding tetrahydrofuran into the reacted solution, wherein the mass ratio of the reacted solution to the tetrahydrofuran is 4: 1, precipitating for 2h, filtering, washing a product with deionized water, and drying in vacuum at 50 ℃ for 30h to obtain an isocyanate-terminated zwitterionic compound;
(D) ultrasonically cleaning a sampling bottle for 10min by using a mixed solution of absolute ethyl alcohol and deionized water with the volume ratio of 1:1, washing by using deionized water, and then using N2Drying, then immersing the cleaned sampling bottle body into a toluene solution of 10 mass percent of 3-aminopropyltriethoxysilane, reacting for 4 hours at 50 ℃, cleaning with absolute ethyl alcohol and deionized water, and cleaning with N2Drying to obtain a sampling bottle body with an aminated surface;
(E) dissolving an isocyanate-terminated zwitterionic compound in tetrahydrofuran, adding N, N-diisopropylethylamine, and uniformly stirring to obtain a zwitterionic compound solution, wherein the mass fraction of the isocyanate-terminated zwitterionic compound is 8%, and the mass fraction of the N, N-diisopropylethylamine is 0.3%; immersing the sampling bottle body with aminated surface into the zwitterionic compound solution for reaction for 1h, taking out, cleaning with tetrahydrofuran, and washing with N2After drying, the sample is heat treated at 100 ℃ for 2h and cooled to obtain the sample bottle treated by antimicrobial adhesion.
The rest is the same as in example 2.
Example 6:
the sampling vial of example 6 was treated for antimicrobial adherence prior to sampling, the treatment comprising the steps of:
(A) mixing the components in a molar ratio of 2.5: 1, mixing toluene diisocyanate and N-methyldiethanolamine, and adding the mixture into N, N-dimethylformamide, wherein the mass ratio of the N, N-dimethylformamide to the toluene diisocyanate is 4: 1, stirring and reacting at 80 ℃ for 12 hours under the protection of nitrogen to obtain a tertiary amine intermediate solution;
(B) dissolving 1, 3-propane sultone in N, N-dimethylformamide to obtain a sultone solution, wherein the mass ratio of the 1, 3-propane sultone to the N, N-dimethylformamide is 1: 3; dropwise adding the sultone solution into the tertiary amine intermediate solution to ensure that the molar ratio of the 1, 3-propane sultone to the N-methyldiethanolamine is 2: 1, stirring and reacting for 20 hours at 100 ℃ under the protection of nitrogen;
(C) adding tetrahydrofuran into the reacted solution, wherein the mass ratio of the reacted solution to the tetrahydrofuran is 6: 1, precipitating for 4h, filtering, washing a product with deionized water, and drying in vacuum at 60 ℃ for 20h to obtain an isocyanate-terminated zwitterionic compound;
(D) ultrasonically cleaning a sampling bottle for 15min by using a mixed solution of absolute ethyl alcohol and deionized water in a volume ratio of 1:2, washing with deionized water, and then cleaning with N2Drying, then immersing the cleaned sampling bottle body into a toluene solution of 3-aminopropyltriethoxysilane with the mass fraction of 15%, reacting for 2 hours at 60 ℃, cleaning with absolute ethyl alcohol and deionized water, and cleaning with N2Drying to obtain a sampling bottle body with an aminated surface;
(E) dissolving an isocyanate-terminated zwitterionic compound in tetrahydrofuran, adding N, N-diisopropylethylamine, and uniformly stirring to obtain a zwitterionic compound solution, wherein the mass fraction of the isocyanate-terminated zwitterionic compound is 12%, and the mass fraction of the N, N-diisopropylethylamine is 0.5%; immersing the sampling bottle body with aminated surface into the zwitterionic compound solution for reaction for 3h, taking out, cleaning with tetrahydrofuran, and washing with N2After drying, the sample is heat treated at 120 ℃ for 1h and cooled to obtain the sample bottle body after the antimicrobial adhesion treatment.
The rest is the same as in example 3.
Comparative example 1:
the sampling bottle in comparative example 1 was not provided with a conical filter, and the rest of the structure and parameters were the same as those in example 1.
Comparative example 2:
the angle between the hypotenuse line and the center line of the conical filter screen in the sampling bottle of comparative example 2 was 16 °, and the rest was the same as in example 1.
Comparative example 3:
the diameter of the top surface of the conical filter in the sampling bottle of comparative example 3 was 15mm, and the rest was the same as in example 1.
Comparative example 4:
the diameter of the top surface of the conical filter in the sampling bottle of comparative example 4 was 30mm, and the rest was the same as in example 1.
Comparative example 5:
the height of the air inlet portion in the sampling bottle in comparative example 5 was 15mm, and the rest was the same as in example 1.
Comparative example 6:
the inside diameter of the cylindrical portions of the inlet portion and the collecting portion of the sampling bottle in comparative example 6 was 46mm, and the rest was the same as in example 1.
Comparative example 7:
the area of the air inlet in the sampling bottle body in comparative example 7 was 230mm2The rest is the same as in example 1.
The sampling bottles of the above examples and comparative examples were tested for their anti-microbial adhesion performance and their sampling efficiency, and the results are shown in table 1.
Wherein, when testing the antimicrobial adhesive property, the test method is as follows:
(1) preparing a bacterial suspension: inoculating the strain into sterilized nutrient broth under aseptic condition, and culturing at 37 deg.C for 24 hr to obtain concentrated bacterial liquid; inoculating the concentrated bacterial liquid to nutrient agar plate with inoculating loop, culturing at 37 deg.C for 24 hr, selecting single colony produced well, inoculating to nutrient broth culture medium, culturing under the same condition for 48 hr, and adjusting the bacterial suspension concentration to 1 × 10 with sterilized normal saline9CFU/L for standby;
(2) bacterial adhesion: cleaning sampling bottle with 75% ethanol, sterilizing by ultraviolet, placing into 200mL sterile nutrient broth, and adding 2mL sterile nutrient broth with concentration of 1 × 109Slightly shaking the CFU/L bacterial suspension, culturing at 37 ℃ for 24h, taking out, washing the sampling bottle with sterile phosphate buffer solution for 3 times, and removing non-adhered bacteria;
(3) observing the adhesion condition of bacteria: slightly heating the sampling bottle body adhered with the bacteria on flame to fix the bacteria, and then dyeing for 1min by using crystal violet dye solution (1g of crystal violet is dissolved in 20mL of 95% ethanol); washing with ultrapure water, drying at 37 deg.C, and observing bacterial adhesion with microscope;
(4) and (3) determining the bacteriostatic condition of the sampling bottle: and (3) measuring the concentration of bacteria in the nutrient broth bacterial suspension cultured in the step (2) after being contacted with the sampling bottle body by using a spectrophotometry method.
The method for measuring the sampling efficiency of the microbial aerosol particles comprises the following steps: in an indoor space of 60 cubic meters, 10 is sprayed using a humidifier6Copies/ml of pseudovirus, gas collection was performed using the sampling bottles of examples and comparative examples, sampling conditions: the gas flow rate is 200L/min, the collection liquid is 15mL (Sutai mechanical 20200073, Jiangsu Mule Biotech, Ltd.), and the sampling time is 20 min. After the collection is finished, the collected collection liquid is concentrated to 3ml by adopting a column method, and the pseudovirus in the collection liquid is quantitatively detected by adopting a PCR instrument.
Table 1: aerosol particle sampling efficiency and antimicrobial adhesion performance test results of the sampling bottles.
As can be seen from table 1, the sampling bottles with the structures and the size parameters of the present invention in examples 1 to 3 have higher sampling efficiency, and the sampling efficiency is lower than that in example 1 when the conical filter net is not disposed in the sampling bottle in comparative example 1 or when the size parameters of each part of the sampling bottle are changed in comparative examples 2 to 7. The sampling bottle provided by the invention is proved to be capable of effectively reducing escape of secondary aerosol and improving sampling efficiency.
In addition, in the embodiments 4 to 6, after the surface antimicrobial adhesion treatment is performed on the sampling bottle body by using the method provided by the invention, the adhesion of microorganisms in the sampling bottle body can be effectively reduced, the sampling efficiency is improved, and meanwhile, the influence of the adhesion of particles on the detection result and the cross contamination on the subsequent sampling are avoided. Moreover, as can be seen from table 1, the concentrations of live bacteria in the bacterial suspensions after the bacterial suspensions are contacted with the sampling bottles subjected to the antimicrobial adhesion treatment in the embodiments 4 to 6 and the bacterial suspensions after the bacterial suspensions are not contacted with the sampling bottles subjected to the antimicrobial adhesion treatment in the embodiments 1 to 3 are not reduced, which proves that the antimicrobial adhesion treatment method of the invention can not inhibit the growth of microorganisms, and can not affect the collected microbial aerosol samples while avoiding the adhesion of particles to the sampling bottles.
Claims (10)
1. A microbial aerosol sampling bottle comprises a sampling bottle body (1) and a sampling bottle cap (2) arranged at the top of the sampling bottle body, and is characterized in that the sampling bottle body sequentially comprises an air inlet part (101), a collecting part (102) communicated with the air inlet part and a placing part (103) which is positioned at the bottom of the collecting part and used for stably placing the sampling bottle, a threaded connection part (104) used for being connected with the sampling bottle cap is arranged on the outer side of the bottle body between the air inlet part and the collecting part, and the sampling bottle cap is sleeved on the outer side of the air inlet part and is in threaded connection with the threaded connection part; be equipped with air inlet (105) on the lateral wall of air inlet portion, be equipped with toper filter screen (4) in the sampling bottle, the toper filter screen passes through filter screen retainer plate (3) and fixes the top at air inlet portion, the filter screen retainer plate is including being used for with air inlet portion top fixed connection's fixed part (301) and being located outlet duct (302) of fixed part below, be equipped with on the fixed part with connecting hole (303) of outlet duct intercommunication, the top fixed connection of outlet duct bottom and toper filter screen.
2. A microbial aerosol sampling bottle according to claim 1, wherein the bottom of the collecting part is inverted conical, the top of the collecting part is cylindrical, and the conical part of the collecting part has the same taper as the conical filter screen.
3. A microorganism aerosol sampling bottle according to claim 1 or 2, wherein the included angle between the inclined side line and the central line of the conical filter screen is 10-15 degrees, and the mesh number of the conical filter screen is 40-100 meshes.
4. A microbial aerosol sampling bottle according to claim 2, wherein the inner diameter of the cylindrical part of the gas inlet part and the cylindrical part of the collecting part is 34-44 mm; the height of the cylindrical part of the air inlet part and the cylindrical part of the collecting part are respectively 20-30 mm and 33-42 mm.
5. A microbial aerosol sampling bottle according to claim 1 or 4, wherein the ratio of the diameter of the top surface of the conical filter screen to the inner diameter of the air inlet portion is 1: (1.6-2.1).
6. A microbial aerosol sampling bottle according to claim 1 or 2, wherein the area of the air inlet on the air inlet part is 162-220 mm2。
7. A microbial aerosol sampling bottle according to claim 1, wherein the size of the fixing part of the filter screen fixing ring is matched with the inner diameter of the air inlet part of the sampling bottle body, and the fixing part of the filter screen fixing ring is clamped in the air inlet part.
8. A microbial aerosol sampling bottle according to claim 2, wherein the cylindrical part of the outer wall of the collecting part is provided with a positioning rib (106) along the height direction of the sampling bottle body, and the positioning rib is provided with a first positioning groove (107).
9. A microbial aerosol sampling bottle according to claim 1, wherein the placing part is provided with a second positioning groove (108).
10. A microbial aerosol sampling bottle according to claim 1, wherein the sampling bottle body is made of glass, and the sampling bottle body is treated with antimicrobial adhesion, and the treatment method comprises the following steps:
(A) the molar ratio is (2-2.5): 1, mixing toluene diisocyanate and N-methyldiethanolamine, adding the mixture into N, N-dimethylformamide, wherein the mass ratio of the N, N-dimethylformamide to the toluene diisocyanate is (3-4): 1, stirring and reacting at 60-80 ℃ for 12-24 hours under the protection of nitrogen to obtain a tertiary amine intermediate solution;
(B) dissolving 1, 3-propane sultone in N, N-dimethylformamide to obtain a sultone solution, wherein the mass ratio of the 1, 3-propane sultone to the N, N-dimethylformamide is 1: (2-3); dropwise adding a sultone solution into a tertiary amine intermediate solution to enable the molar ratio of 1, 3-propane sultone to N-methyldiethanolamine to be (1-2): 1, stirring and reacting for 20-30 h at 80-100 ℃ under the protection of nitrogen;
(C) adding tetrahydrofuran into the reacted solution, wherein the mass ratio of the reacted solution to the tetrahydrofuran is (4-6): 1, precipitating for 2-4 h, filtering, washing a product with deionized water, and drying in vacuum at 50-60 ℃ for 20-30 h to obtain an isocyanate-terminated zwitterionic compound;
(D) ultrasonically cleaning a sampling bottle for 10-15 min by using a mixed solution of absolute ethyl alcohol and deionized water with the volume ratio of 1 (1-2), washing by using deionized water, and then using N2Drying, then immersing the cleaned sampling bottle body into a toluene solution of 10-15% by mass of 3-aminopropyltriethoxysilane, reacting for 2-4 h at 50-60 ℃, cleaning with absolute ethanol and deionized water, and cleaning with N2Drying to obtain a sampling bottle body with an aminated surface;
(E) dissolving an isocyanate-terminated zwitterionic compound in tetrahydrofuran, adding N, N-diisopropylethylamine, and uniformly stirring to obtain a zwitterionic compound solution, wherein the mass fraction of the isocyanate-terminated zwitterionic compound is 8-12%, and the mass fraction of the N, N-diisopropylethylamine is 0.3-0.5%; immersing the sampling bottle body with the aminated surface into a zwitterionic compound solution for reaction for 1-3 h, taking out, cleaning with tetrahydrofuran, and using N2And (3) after drying, carrying out heat treatment at 100-120 ℃ for 1-2 h, and cooling to obtain the sampling bottle body subjected to antimicrobial adhesion treatment.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090193971A1 (en) * | 2008-01-04 | 2009-08-06 | The Texas A&M University System | Advanced wetted wall aerosol sampling cyclone system and methods |
| CN110376027A (en) * | 2019-07-26 | 2019-10-25 | 东南大学 | Multi-stage biological aerosol sampler and the method for sampling |
| CN111500427A (en) * | 2020-03-21 | 2020-08-07 | 深圳市朗司医疗科技有限公司 | Portable wet wall cyclone microorganism aerosol collector |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090193971A1 (en) * | 2008-01-04 | 2009-08-06 | The Texas A&M University System | Advanced wetted wall aerosol sampling cyclone system and methods |
| CN110376027A (en) * | 2019-07-26 | 2019-10-25 | 东南大学 | Multi-stage biological aerosol sampler and the method for sampling |
| CN111500427A (en) * | 2020-03-21 | 2020-08-07 | 深圳市朗司医疗科技有限公司 | Portable wet wall cyclone microorganism aerosol collector |
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