CN102495051A - Device and method for quickly detecting biological activity and metabolism - Google Patents
Device and method for quickly detecting biological activity and metabolism Download PDFInfo
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- CN102495051A CN102495051A CN2011103928136A CN201110392813A CN102495051A CN 102495051 A CN102495051 A CN 102495051A CN 2011103928136 A CN2011103928136 A CN 2011103928136A CN 201110392813 A CN201110392813 A CN 201110392813A CN 102495051 A CN102495051 A CN 102495051A
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/24—Gas permeable parts
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/38—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of metabolites or enzymes in the cells
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
Abstract
The invention belongs to the technical field of biological activity or metabolism detection, and particularly relates to a device and a method for quickly detecting the biological activity and the metabolism. The device is a sealed container, and separated two cavities or more are arranged inside the sealed container. Breathable material layers containing indicating agents are arranged in the cavities, and the indicating agents are used for indicating and detecting conditions of growth and metabolism of living beings. When the indicating agents in different cavities are same and thicknesses of breathable material layers are different, quick detection of phase difference can be achieved. When the indicating agents in different cavities are different and thicknesses of breathable material layers are different, different metabolites can be synchronously detected. The device and the method for quickly detecting the biological activity and the metabolism can be used for relative detection and quarantine inspection of culture of animals, plants, cells and microorganisms and achieve automatic monitoring of various biological tests.
Description
Technical field
The invention belongs to biologically active or metabolism detection technique field, be specifically related to a kind of apparatus and method that are used for fast detecting biologically active, metabolism.
Background technology
Can produce metabolic product gas in most of biological growths, the metabolic process, the gas of release comprises: oxygen (O
2), carbon dioxide (CO
2), ammonia (NH
3) or sulfuretted hydrogen (H
2S) etc.
Preponderate by photosynthesis under light conditions for plant (comprising unicellular lower eukaryotes such as algae), mainly is to absorb CO
2, discharge O
2, the unglazed photograph under the condition only carried out respiratory metabolism, absorbs O2, discharges CO
2, therefore, through CO
2, but O2 concentration detects monitoring plant growth metabolism situation.
In the cellular incubation, cell consumption glucose can discharge CO
2, CO
2Be sour gas, make nutrient solution be acid, usually use phenol redly, phenol redly explain that by rose-red (alkaline look) flavescence (sour look) cell grows as the pH indicator.
Many growth of microorganism metabolism respiratories are also similar with cell, can discharge CO
2, for this reason, through measuring the CO that discharges
2Amount (methods such as pH photon or electronics) is monitored, whether is measured growth of microorganism, growth characteristics, the record growth curve.
Some microorganisms contain urase can digest urea generation ammonia (NH
3), ammonia is an alkaline gas, can use pH indicator such as bromthymol blue, becomes blue (alkaline look) by yellow (acid look) and detects.
In addition, some bacterium can be decomposed sulfur-containing amino acid generation sulfuretted hydrogen (H in the nutrient culture media
2S), sulfuretted hydrogen is stronger reductive agent, can use redox indicator (like lead acetate) colour developing reflection growth of microorganism situation.
Because there are certain toxicity in pH indicator, redox indicator pair cell, bacterial growth; Even can suppress growth metabolism; All adopt out-phase to detect at present; Be indicator and cultivate not mutually same, indicator (phase) is when color changes, through the change acquisition biological growth data of light emitting diode emission-acceptances detection indicator (phase) wavelength.As shown in Figure 1, the problem of its existence:
1, only an indicator can only detect a kind of metabolic product mutually, can not two or more metabolic products of synchronous monitoring.
2, because gas entering indicator mutual-assistance indicator phase change is the process of a gradual change from top to bottom; Present method must be waited until the generation that could survey growth or metabolism when bottom colors changes, and the time delay of detection has reduced the sensitivity that detects undoubtedly and reduced.
3, a lot of especially severe bacterias of Clinical microorganism, slowly the bacterium (comprising mould, saccharomycete, Much's bacillus etc.) that grows detects; The existence of microorganism is discerned, judged to testing goal often; Need obtain cultivation results (positive still negative) as early as possible; But the detection receptacle of using at present only provides an indicator phase, owing to difference between indicator itself, the indicator phase, forces the baseline of instrument need set than high parameter; The directly sensitivity of influence detection has increased and has estimated the difficulty of differentiating the result.
Summary of the invention
The object of the present invention is to provide a kind of simple in structure, monitor the apparatus and method of fast detecting biologically active, metabolism easily.
The device of fast detecting biologically active provided by the invention, metabolism is an airtight container, and the cavity of separation is contained in its inboard, and said cavity can be Room 2, also can be for multicell (more than the Room 2, like Room 3, Room 4 etc.), shown in Fig. 2 a, b, c, d.
Among the present invention, be provided with the breathable material layer that contains indicator in the said cavity; The thickness of the breathable material layer in the different cavitys can be identical, also can be different; The kind of contained indicator can be identical in the breathable material layer in the different cavitys, also can be different.Condition of different can be used for the different detection purpose.For example:
Contained indicator kind is identical in the breathable material layer in the different cavitys, but the gas permeable material layer thickness is different, differs fast detecting in the time of can realizing.
Contained indicator kind is different in the breathable material layer in the different cavitys, and the gas permeable material layer thickness is also different, can be used for the different metabolic product of synchronous detection.
Among the present invention, the cavity of separation can be located at the inboard bottom of device (container), top (in the lid) or side etc., shown in Fig. 2 e, f, g.
Among the present invention, when the gas permeable material layer thickness was different, the thickness ratio can be between 1:2 ~ 10.
Among the present invention, said indicator is meant can be with O
2, CO
2, NH
3, H
2Gas concentrations such as S change and the material that changes, and said variation can be but be not limited to absorbing light or wavelength of transmitted light changes.Indicator can be but be not limited to pH indicator, redox indicator etc.
Among the present invention, the said breathable material layer that contains indicator, refer to through means such as absorption, emulsification, dissolving with said indicator solid phase in gas permeable material; Gas permeable material can be but be not limited to Electrostatic Absorption nylon membrane, cellulose, agar, silicon rubber, resin etc.
Among the present invention, said airtight container is with gas impermeable material manufacturing such as transparent, translucent or opaque; Its material can be but be not limited to PC, PP, PS, PE, PTFE, glass etc. and can guarantee from the container External Observation or detect in the container to change (absorbing light or wavelength of transmitted light change).
Utilize said apparatus of the present invention can the fast detecting biologically active, the concrete condition of metabolism; Concrete steps do; Culture is positioned in the airtight container of the present invention,, can judges through range estimation according to absorbing light (color, gray scale) or the wavelength of transmitted light change or the difference of indicator; Also can be connected with single-chip microcomputer, computing machine through modes such as scanning, shooting, photograph; Through software monitoring, analyze, judge, record growth curve and result, thereby realize synchronously, the time differ fast detecting biologically active, metabolism situation, see Fig. 3, shown in Figure 4.For example:
Contained indicator kind is identical in the breathable material layer in the different cavitys, but the gas permeable material layer thickness differs fast detecting when different in the time of can realizing.
Contained indicator kind is different in the breathable material layer in the different cavitys, and the gas permeable material layer thickness also not simultaneously, can be used for the different metabolic product of synchronous detection.
Among the present invention, cavity design is had versatility in the container cover inboard, not only can be used for liquid culture and detect, also can be used for solid culture and detect, can indicate the biological growth in solid inclined-plane.
Device of the present invention can be as required; Design is suitable for container, culture flask or the culture tube etc. of the sealing of different sizes, shape arbitrarily; Can be used for animal, plant, cell, microbe culture relevant check, quarantine, and the tests such as susceptibility of various microorganisms.
Use apparatus of the present invention and method can but be not limited to be used for the associated gas of generations such as fast detecting biology, physics, chemistry.
Description of drawings
Fig. 1 is the biological metabolism detection method diagram of prior art.
The inboard cavity body structure diagram of the airtight container that is used for quick indicator organism metabolism that Fig. 2 designs for the present invention.Wherein, (a) for 2 chamber bodies (concentric circles) are arranged, (b) for 2 chamber bodies (dividing half-and-half separated) are arranged, (c) being 3 chamber bodies, (d) is 4 chamber bodies; (e) for there being 2 chamber bodies to be located at container top (lid) inboard, (f) for there being 2 chamber bodies to be located at the container bottom inboard, (g) for there being 2 chamber bodies to be located at the container side inboard.
The cover for sealed container structure of Fig. 3 for using in the embodiment of the invention.Wherein, the concentric circles 2 chamber bodies design stereographic map that (a) contains for lid is inboard (b) is cross-sectional structure.
Fig. 4 is the embodiment of the invention 1 diagram, places the indicator (phase) of identification gas with various in the 2 chamber bodies of cover for sealed container inboard respectively, can survey the diagram of gas with various simultaneously.
Fig. 5 is the embodiment of the invention 2,3 diagrams, place identical indicator (embodiment 2 uses fluorescent pH indicators, and embodiment 3 uses the pH colored indicators) in the 2 chamber bodies of cover for sealed container inboard, but the gas permeable material layer thickness is different.
Fig. 6 is the embodiment of the invention 3 results diagrams, the breathable material layer (measure CO that contains the pH colored indicator of placing different-thickness in the 2 chamber bodies of cover for sealed container inboard
2Burst size), be no bacterial growth from the result (a) of bottle cap top observation, indicate with consistent with reference to phase color, gray scale, (b) for bacterial growth is arranged, indicate with inconsistent with reference to color mutually, gray scale; (c) be no bacterial growth result, (d) the bacteria growing result arranged for scanning from the bottle cap top from bottle cap top scanning; The metabolism intensity curve of sweep record from the bottle cap top: (e) no growth curve of bacteria, (f) for the bacteria growing curve is arranged.
Fig. 7 differs the method for quick principle when of the present invention and further describes.
Embodiment
The invention is further illustrated by the following examples.
In the embodiment of the invention, it is inboard that cavity is located at the airtight container lid, and lid is inboard to form 2 chamber bodies with two inside and outside circles of concentric circles, as shown in Figure 3.(a) show the spatial structure that concentric circles 2 chamber bodies cover, (b) show cross section plane figure.
Nutrient culture media adopts the general enriched medium that Shanghai Jicai Medical Instruments Co., Ltd. produces among the embodiment, and prescription (calculating by 1000mL) is:
| Michaelis 7H9 meat soup | 5.9g |
| BSA | 5.0g |
| Glucose | 2.0g |
| Catalase | 0.003g |
| Glycerine | ? 3 mL |
| Purified water | 1000 mL |
| Add up to | 1000 mL |
Under the aseptic condition with nutrient culture media with every bottle of 5ml packing.
Embodiment 1
Place respectively in the 2 chamber bodies in the above-mentioned bottle cap and can discern carbon dioxide (CO
2) (pH indicator) and sulfuretted hydrogen (H
2S) breathable material layer of (lead acetate oxygenant), the carbon dioxide (CO that the metabolism of synchronous detection biological growth discharges
2) and sulfuretted hydrogen (H
2S) (Fig. 4).
1.pH indicator prepares mutually:
A. get thymol blue (Thymol blue pKa=8.9) 0.1g, add 1N NaOH 2ml dissolving, it is subsequent use to add water to 10ml.
B. get silastic liquid (GE 2501) 10g, add above-mentioned solution 0.2ml, after the mixing, inject respective chamber, cold curing 24 hours, it is blue that the silica gel after the curing is.
2. sulfuretted hydrogen (H
2S) indication preparation mutually:
A. get lead acetate 0.1g, add water 9ml dissolving, it is subsequent use to add 10% TW-80 1ml.
B. get silastic liquid (GE 2501) 10g, add above-mentioned solution 0.2ml, after the mixing, inject respective chamber, cold curing 24 hours, the silica gel after the curing is white in color.
Pb
2+?+S
2-===?PbS?↓?。
3. get the culture tube that two pipes contain nutrient culture media, the crooked bacterium (Campylobacter fetus ATCC 27374) of inoculation fetus is cooked contrast for one, puts 37 degree and cultivates 24hr.
4. result: like Fig. 4 (b), the phase transformation of pH indicator is yellow, sulfuretted hydrogen (H
2S) indication turns black mutually, explains that this bacterium produces H in growth
2S; And control tube does not have any variation, and two keep original tone mutually.
Place the ventilative silastic-layer of different-thickness in the above-mentioned bottle cap in the 2 chamber bodies respectively, in ventilative silastic-layer, contain and to discern carbon dioxide (CO
2) the fluorescence indicator 8-hydroxyl pyrene-1,3 of gas, 6-trisulfonic acid trisodium salt (HPTS); This HPTS is pH fluorescence indicator commonly used; Pka=7.3, maximum excitation wavelength 405nm/455nm changes the carbon dioxide that the detection of biological growth metabolism produces through fluorescence intensity.
1.pH fluorescence indicator prepares mutually:
A.8-hydroxyl pyrene-1,3,6-trisulfonic acid trisodium salt 0.1g adds 1N NaOH 1ml dissolving, and it is subsequent use to add water to 10ml.
B. get silastic liquid (GE 2501) 10g, add above-mentioned solution 0.2ml, after the mixing, inject respective chamber, indication is 1 thickness 1mm mutually, with reference to phase 2 thickness 5mm, cold curing 24 hours (like Fig. 5).
2. get the culture tube that two pipes contain nutrient culture media, an inoculation mycobacterium smegmatis (Mycobacterium smegmatis MC2 155, ATCC 700084) is done contrast for one, puts 37 degree and cultivates.
3. (control tube fluorescence remains unchanged the result for 3W, 450nm) observed result, and inoculated tube is inoculated back 5 hours indication phase fluorescence intensities and significantly changed, and with reference to phase fluorescence intensity changes needs 7 hours at the LED uviol lamp.
Place the silastic-layer of different-thickness in the above-mentioned bottle cap in the 2 chamber bodies respectively, contain at silastic-layer and can discern carbon dioxide (CO
2) indicator to xylenol blue (Xylenol blue pka=8.9), the silastic-layer collocation method that contains indicator is following:
1.pH fluorescence indicator prepares mutually:
A. get xylenol blue 0.1g, add 1N NaOH 2ml dissolving, it is subsequent use to add water to 10ml.
B. get silastic liquid (GE 2501) 10g, add above-mentioned solution 0.2ml, after the mixing, inject respective chamber, indication is 1 thickness 1mm mutually, with reference to phase 2 thickness 6mm, cold curing 24 hours (like Fig. 5).
2. get the culture tube that two pipes contain nutrient culture media, an inoculation mycobacterium smegmatis (Mycobacterium smegmatis MC2 155, ATCC 700084) is done contrast for one, puts 37 degree and cultivates.
3. result: control tube two remains consistent blue mutually, and inoculated tube is inoculated back 6 hours, indicates mutually that blueness begins to shoal, Huang, and obviously distinguishes with reference to being on good terms, and needs 8 hours with reference to flavescence fully mutually (mutually identical with indication).
Among the present invention; The difference of the color of bottle cap internal indicator (gray scale) can be estimated judgement (a, b among Fig. 6); Also can be connected with single-chip microcomputer, computing machine through modes such as scanning, shooting, photograph; Through software analysis, judgement (c, d among Fig. 6), or monitor, write down growth curve (e, f among Fig. 6) in real time.
" time phase difference detection method " according to the invention refer to based on indication with differ when intensity variation exists mutually, utilize light intensity difference to differentiate the growing point method of biology.If will be with reference to being set at existing method mutually, the inventive method is interpretation biological growth, activity or metabolism (Fig. 7) earlier.
Claims (10)
1. the device of biologically active and metabolism fast detecting is characterized in that being a kind of airtight container, and the cavity of separation is contained in its inboard, and said cavity is Room 2, perhaps is the above multicells in Room 2; Be provided with the breathable material layer that contains indicator in the said cavity; The thickness of the breathable material layer in the different cavitys is identical or different; Contained indicator kind is identical or different in the breathable material layer in the different cavitys.
2. device according to claim 1 is characterized in that the cavity of said separation is located at airtight container inboard bottom, top or side.
3. device according to claim 1 is characterized in that contained indicator kind is identical in the breathable material layer in the said different cavity, and the gas permeable material layer thickness is different.
4. device according to claim 1 is characterized in that contained indicator kind is different in the breathable material layer in the said different cavity, and the gas permeable material layer thickness is different.
5. according to claim 3 or 4 described devices, when it is characterized in that said gas permeable material layer thickness is different, its thickness is than between 1:2 ~ 10.
6. device according to claim 1 is characterized in that said indicator is for can be with O
2, CO
2, NH
3, H
2The S gas concentration changes and the material that changes.
7. device according to claim 6 is characterized in that said indicator is pH indicator or redox indicator.
8. device according to claim 1 is characterized in that the said breathable material layer that contains indicator, through absorption, emulsification or dissolving said indicator solid phase is obtained in gas permeable material; The material of said breathable material layer is nylon membrane, cellulose, agar, silicon rubber or resin; The material of said airtight container is transparent, translucent or opaque gas impermeable material.
9. one kind is utilized the said device of one of claim 1-8 to carry out the method that biologically active and metabolism detect; It is characterized in that concrete steps are: culture is positioned in the airtight container of the present invention; According to the requirement that detects, the thickness of breathable material layer and the kind of indicator in the number of the cavity that selection is separated, each cavity, the absorbing light of indicator or the change of wavelength of transmitted light or difference; Judge through range estimation; Perhaps be connected with single-chip microcomputer, computing machine through scanning, shooting or photographic means, through software monitoring, analysis, judgement, record growth curve and result, thereby detection of biological is active, the metabolism situation.
10. method according to claim 9 is characterized in that said culture is liquid culture or solid culture; Be applicable to check, quarantine that animal, plant, cell, microbe culture are correlated with, and the drug sensitive test of various microorganisms.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102816688A (en) * | 2012-09-14 | 2012-12-12 | 江苏嘉语生物医药技术有限公司 | Microorganism culture detection device in oxygen-controllable environment and detection method using same |
| CN103784145A (en) * | 2014-01-22 | 2014-05-14 | 中国农业科学院饲料研究所 | Seven-channel small animal respiratory metabolism measuring device |
| CN103923826A (en) * | 2014-04-30 | 2014-07-16 | 上海市肺科医院 | Detector tube for bacterial culture |
| CN104797925A (en) * | 2012-11-09 | 2015-07-22 | 弗劳恩霍夫应用研究促进协会 | Receptacle and system for optically analyzing a sample without optical lenses |
| CN105209911A (en) * | 2013-03-11 | 2015-12-30 | 贝克顿·迪金森公司 | Bacterial detection cartridge |
| CN106932384A (en) * | 2015-12-29 | 2017-07-07 | 深圳市芭田生态工程股份有限公司 | Agar indicator, its preparation method and application |
| CN107988075A (en) * | 2018-01-04 | 2018-05-04 | 广东省微生物研究所（广东省微生物分析检测中心） | A kind of method that screening from water body produces the strain of high activity bacterium volatile matter |
| WO2021077456A1 (en) * | 2019-10-24 | 2021-04-29 | 中山大学 | Filter device and method used for microbial enzymatic activity testing |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102816688B (en) * | 2012-09-14 | 2013-11-20 | 江苏嘉语生物医药技术有限公司 | Microorganism culture detection device in oxygen-controllable environment and detection method using same |
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| CN104797925A (en) * | 2012-11-09 | 2015-07-22 | 弗劳恩霍夫应用研究促进协会 | Receptacle and system for optically analyzing a sample without optical lenses |
| US9909162B2 (en) | 2013-03-11 | 2018-03-06 | Becton, Dickinson And Company | Bacterial detection cartridge |
| US10557163B2 (en) | 2013-03-11 | 2020-02-11 | Becton Dickinson And Company | Bacterial detection cartridge |
| CN105209911A (en) * | 2013-03-11 | 2015-12-30 | 贝克顿·迪金森公司 | Bacterial detection cartridge |
| CN103784145A (en) * | 2014-01-22 | 2014-05-14 | 中国农业科学院饲料研究所 | Seven-channel small animal respiratory metabolism measuring device |
| CN103923826B (en) * | 2014-04-30 | 2016-04-06 | 上海市肺科医院 | A kind of microbial culture detector tube |
| CN103923826A (en) * | 2014-04-30 | 2014-07-16 | 上海市肺科医院 | Detector tube for bacterial culture |
| CN106932384A (en) * | 2015-12-29 | 2017-07-07 | 深圳市芭田生态工程股份有限公司 | Agar indicator, its preparation method and application |
| CN107988075A (en) * | 2018-01-04 | 2018-05-04 | 广东省微生物研究所（广东省微生物分析检测中心） | A kind of method that screening from water body produces the strain of high activity bacterium volatile matter |
| CN107988075B (en) * | 2018-01-04 | 2021-01-08 | 广东省微生物研究所（广东省微生物分析检测中心） | Method for screening strains generating high-activity bacterial volatile matters from water body |
| WO2021077456A1 (en) * | 2019-10-24 | 2021-04-29 | 中山大学 | Filter device and method used for microbial enzymatic activity testing |
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Application publication date: 20120613 |