CN107603858A - The preparation of anaerobe culture medium, inoculation and culture apparatus - Google Patents
The preparation of anaerobe culture medium, inoculation and culture apparatus Download PDFInfo
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- 238000011081 inoculation Methods 0.000 title claims abstract description 36
- 239000001963 growth medium Substances 0.000 title claims abstract description 34
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000005273 aeration Methods 0.000 claims abstract description 17
- 238000010438 heat treatment Methods 0.000 claims abstract description 12
- 239000003365 glass fiber Substances 0.000 claims abstract description 10
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229910052751 metal Inorganic materials 0.000 claims abstract description 4
- 239000002184 metal Substances 0.000 claims abstract description 4
- 238000004804 winding Methods 0.000 claims abstract description 4
- 230000001012 protector Effects 0.000 claims abstract 2
- 229910001220 stainless steel Inorganic materials 0.000 claims description 28
- 239000010935 stainless steel Substances 0.000 claims description 24
- 241001290534 Lachnocaulon Species 0.000 claims description 18
- 229910000831 Steel Inorganic materials 0.000 claims description 12
- 239000010959 steel Substances 0.000 claims description 12
- 239000011521 glass Substances 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 3
- 238000007667 floating Methods 0.000 claims description 2
- 239000007789 gas Substances 0.000 description 75
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 26
- 239000000243 solution Substances 0.000 description 22
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 20
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 18
- 229910002092 carbon dioxide Inorganic materials 0.000 description 16
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 12
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 11
- 229910052757 nitrogen Inorganic materials 0.000 description 11
- 229940088594 vitamin Drugs 0.000 description 9
- 235000013343 vitamin Nutrition 0.000 description 9
- 239000011782 vitamin Substances 0.000 description 9
- 229930003231 vitamin Natural products 0.000 description 9
- 150000003722 vitamin derivatives Chemical class 0.000 description 9
- 239000001569 carbon dioxide Substances 0.000 description 8
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 8
- 238000003501 co-culture Methods 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 241001135750 Geobacter Species 0.000 description 6
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 6
- 208000006440 Open Bite Diseases 0.000 description 6
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 6
- 239000011573 trace mineral Substances 0.000 description 6
- 235000013619 trace mineral Nutrition 0.000 description 6
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 5
- 239000002054 inoculum Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 241001135751 Geobacter metallireducens Species 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 238000006392 deoxygenation reaction Methods 0.000 description 4
- 229910001873 dinitrogen Inorganic materials 0.000 description 4
- 229910052564 epsomite Inorganic materials 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000009938 salting Methods 0.000 description 4
- GHOKWGTUZJEAQD-SSDOTTSWSA-N 3-[[(2s)-2,4-dihydroxy-3,3-dimethylbutanoyl]amino]propanoic acid Chemical compound OCC(C)(C)[C@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-SSDOTTSWSA-N 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 3
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 description 3
- 229910002566 KAl(SO4)2·12H2O Inorganic materials 0.000 description 3
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 3
- 229910004619 Na2MoO4 Inorganic materials 0.000 description 3
- 229910020350 Na2WO4 Inorganic materials 0.000 description 3
- 150000007513 acids Chemical class 0.000 description 3
- 238000004500 asepsis Methods 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 150000001615 biotins Chemical class 0.000 description 3
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- LBGWCDQCRMORLC-UHFFFAOYSA-N ethanol methane Chemical compound C[H].[H]C([H])[H].[H]CC([H])O LBGWCDQCRMORLC-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229960000304 folic acid Drugs 0.000 description 3
- 235000019152 folic acid Nutrition 0.000 description 3
- 239000011724 folic acid Substances 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 235000019136 lipoic acid Nutrition 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 229960003512 nicotinic acid Drugs 0.000 description 3
- 235000001968 nicotinic acid Nutrition 0.000 description 3
- 239000011664 nicotinic acid Substances 0.000 description 3
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000011684 sodium molybdate Substances 0.000 description 3
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 3
- 239000011655 sodium selenate Substances 0.000 description 3
- XMVONEAAOPAGAO-UHFFFAOYSA-N sodium tungstate Chemical compound [Na+].[Na+].[O-][W]([O-])(=O)=O XMVONEAAOPAGAO-UHFFFAOYSA-N 0.000 description 3
- 235000019157 thiamine Nutrition 0.000 description 3
- 150000003544 thiamines Chemical class 0.000 description 3
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- ALYNCZNDIQEVRV-UHFFFAOYSA-N 4-aminobenzoic acid Chemical compound NC1=CC=C(C(O)=O)C=C1 ALYNCZNDIQEVRV-UHFFFAOYSA-N 0.000 description 2
- 229910021592 Copper(II) chloride Inorganic materials 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241001494297 Geobacter sulfurreducens Species 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 239000007836 KH2PO4 Substances 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 2
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000001530 fumaric acid Substances 0.000 description 2
- 239000008246 gaseous mixture Substances 0.000 description 2
- 239000011565 manganese chloride Substances 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 229910052603 melanterite Inorganic materials 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- LGQLOGILCSXPEA-UHFFFAOYSA-L nickel sulfate Chemical compound [Ni+2].[O-]S([O-])(=O)=O LGQLOGILCSXPEA-UHFFFAOYSA-L 0.000 description 2
- 229910000363 nickel(II) sulfate Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 2
- 239000011686 zinc sulphate Substances 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- 239000004201 L-cysteine Substances 0.000 description 1
- 235000013878 L-cysteine Nutrition 0.000 description 1
- 229910021586 Nickel(II) chloride Inorganic materials 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 238000005276 aerator Methods 0.000 description 1
- 239000005030 aluminium foil Substances 0.000 description 1
- 229960004050 aminobenzoic acid Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 229910052927 chalcanthite Inorganic materials 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 150000001860 citric acid derivatives Chemical class 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229960002413 ferric citrate Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- NPFOYSMITVOQOS-UHFFFAOYSA-K iron(III) citrate Chemical compound [Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NPFOYSMITVOQOS-UHFFFAOYSA-K 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 238000011177 media preparation Methods 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 229910052979 sodium sulfide Inorganic materials 0.000 description 1
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 description 1
- 244000000000 soil microbiome Species 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 229910052720 vanadium Inorganic materials 0.000 description 1
- 239000011592 zinc chloride Substances 0.000 description 1
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of preparation of anaerobe culture medium, inoculation and culture apparatus, including gas bomb group（1）, float gas flowmeter group（4）, quartz glass tube（11）It is sequentially connected with aeration component, the gas bomb group（1）N is respectively provided with for three2、CO2And H2/CO2Gas bomb（1.1、1.2、1.3）；Quartz glass tube（11）Built with copper wire（10）, quartz glass tube（11）Outside winding glass fibre heating wire（9）And have metal protection protector, glass fibre heating wire connects single-phase contact type voltage regulator（19）.Apparatus of the present invention small volume, it is simple to operate, it is cheap, can freely it operate, while meet the preparation, inoculation and culture of anaerobe culture medium, it is applicable various environment, the microculture of various culture atmosphere.
Description
Technical field
The present invention relates to field of microbial culture technology, a kind of specifically laboratory anaerobe culture medium system
Standby, inoculation and culture apparatus.
Background technology
Anaerobe is extensive in distributed in nature, and species is various, and effect also increasingly draws attention.Anaerobe is cultivated
Key be used to such microbial inoculant and culture when all in the environment of anaerobic.
At present, anaerobe culture is typically using alkaline Pyrogallil acid, anaerobic jar cultivation, cooked meat medium
Method and anaerobism glove box.
Alkaline Pyrogallil acid is simple to operate without special and expensive equipment, suitable for any sealable container,
Anaerobic environment can be established rapidly, and its shortcoming is that a small amount of carbon monoxide can be produced in oxidizing process, the life to some anaerobic bacterias
With inhibitory action.
Anaerobic jar can not realize culture medium preparation, the anaerobic environment of microbial inoculant link, can only ensure incubation
Anaerobic environment.
Cooked meat medium method is usually used in the fluid nutrient medium culture of anaerobe, and versatility is poor, can only be applicable spy
Determine the culture of anaerobe.
Though anaerobic culture box can be not only used for anaerobe culture, also can anaerobe inoculation, there is also it is many not
Foot, such as inconvenient for operation, expensive equipment, volume are larger, are not suitable for extreme anaerobe（Such as seabed strain, deep soil bacterium
Kind）Culture medium is unable to constant temperature.
There is also many drawbacks for anaerobism glove box：
（1）Due to some strains need can grow under elevated pressure conditions, be such as derived from seabed strain, be derived from deep soil strain but
Anaerobism glove box can not simulate different pressure conditions, and so far seabed strain and soil strain can not normally give birth in the case of compression
It is long, benefit from limited；
（2）Integrally can not arbitrarily it place, many experiments need to carry out in constant incubator.Therefore, design and prepare a kind of behaviour
Work is simple, cost is low, and can realize the preparation of anaerobe culture medium, is inoculated with and be incubated at integral anaerobic culture device
Great meaning.
The content of the invention
The purpose of the present invention is to overcome the shortcomings of the prior art, there is provided a kind of anaerobe culture medium is prepared, connect
Kind and culture apparatus, the device volume is small, simple to operate, cheap, can freely operate, and is applicable the microorganism training of various environment
Support.
A kind of preparation of anaerobe culture medium, inoculation and culture apparatus, including：Gas bomb group 1, float gas flow
Meter group 4, quartz glass tube 11 and aeration component, the gas bomb group 1 are three and are respectively provided with N2、CO2And H2/CO2Gas
Body steel cylinder（1.1、1.2、1.3）；
The float gas flowmeter group 4 is three float gas flowmeters（4.1、4.2、4.3）One end is divided by stainless steel tube 3
Not with three gas bombs（1.1、1.2、1.3）It is corresponding to be connected, the first float gas flowmeter 4.1 and the second float gas flow
The other end of meter 4.2 is connected with the first cutting sleeve type three-way joint 5, then the port of export and the 3rd of the first cutting sleeve type three-way joint 5
The other end of float gas flowmeter 4.3 is connected with bite type tee ball valve 6, and the port of export of bite type tee ball valve 6 passes through outer
The stainless steel tube for being cased with rubber tube 7 is connected with the bottom of quartz glass tube 11；
Quartz glass tube 11 simultaneously has metal built with copper wire 10, the outside of quartz glass tube 11 winding glass fibre heating wire 9
Protective case, glass fibre heating wire connect single-phase contact type voltage regulator 19；
The top of quartz glass tube 11 is connected by being cased with the stainless steel tube of rubber tube 7 outside with the second cutting sleeve type three-way joint 5.1,
The one outlet end of second cutting sleeve type three-way joint 5.1 is sequentially connected with integrated valve hatpin valve 12, rubber tube 14, glass syringe
17th, stainless steel syringe needle 18；Another port of export of second cutting sleeve type three-way joint 5.1 is connected with integrated valve hatpin valve 12, then
Multiple aeration components are parallel with, are connected between aeration component with the 3rd cutting sleeve type three-way joint 5.2.
Gas bomb outlet is provided with steel cylinder pressure-reducing valve 2.
Stainless steel tube 3 is the stainless steel tube of 1/8 inch of external diameter.
Aeration component is made up of toggle switch valve 13, rubber tube 14, stainless steel syringe needle 15, anaerobism bottle 16.
The float gas flowmeter group 4, cutting sleeve type three-way joint（5、5.1、5.2）, bite type tee ball valve 6 and overall
Formula bonnet needle-valve 12 is installed in same stainless steel surface.
H2/CO2Gas bomb is built with hydrogen and carbon dioxide.
Compared with prior art, the beneficial effects of the present invention are：
1st, two float gas flowmeters are connected using cutting sleeve type three-way joint, the change of Anaerobic culturel atmosphere composition has can be achieved
Change；
2nd, bite type tee ball valve realizes the mutual conversion between different culture atmosphere；
3rd, two integrated valve hatpin valves are connected using cutting sleeve type three-way joint to realize that gas circuit shunts, it is whole by adjusting two
Body formula bonnet needle-valve can realize that anaerobic culture medium prepares operation or individually operated while inoculation with anaerobe；
4th, quartz glass tube, copper wire, glass fibre heating wire and single-phase contact type voltage regulator, the online de- of gas is realized
Oxygen；
Apparatus of the present invention small volume, it is simple to operate, it is cheap, can freely it operate, while meet anaerobe culture medium
Prepare, be inoculated with and cultivate, be applicable various environment, the microculture of various culture atmosphere.
Brief description of the drawings
Fig. 1 is the overall structure diagram of present system.
Fig. 2 a are not to be inoculated with Geobacter metallireducens FC culture mediums；
FC culture mediums after Fig. 2 b are cultivated one week for inoculation Geobacter metallireducens.
Fig. 3 a are not to be inoculated with Geobacter sulfurreducens NBF culture mediums；
NBF culture mediums after Fig. 3 b are cultivated one week for inoculation Geobacter sulfurreducens.
Fig. 4 is the Geobacter metallireducens and co-culture of Methanosarsina barkeri 800
Coupling metabolism ethanol methane phase curve map.
In figure, 1 is gas bomb group, and 1.1 be first gas steel cylinder, and 1.2 be second gas steel cylinder, and 1.3 be third gas
Steel cylinder, 2 be steel cylinder pressure-reducing valve, and 3 be stainless steel tube, and 4 be float gas flowmeter group, and 4.1 be the first float gas flowmeter,
4.2 be the second float gas flowmeter, and 4.3 be the 3rd float gas flowmeter, and 5 be the first cutting sleeve type three-way joint, and 5.1 be
Two cutting sleeve type three-way joints, 5.2 be the 3rd cutting sleeve type three-way joint, and 6 be bite type tee ball valve, and 7 be rubber tube, and 8 be electric wire,
9 be glass fibre heating wire, and 10 be copper wire, and 11 be quartz glass tube, and 12 be integrated valve hatpin valve, and 13 be toggle switch
Valve, 14 be rubber tube, and 15,18 be stainless steel syringe needle, and 16 be anaerobism bottle, and 17 be glass syringe, and 19 be single-phase contact pressure regulation
Device.
Embodiment
The present invention will be further described with embodiment for explanation below in conjunction with the accompanying drawings, but not protects model to it
The limitation enclosed.
As Figure 1-Figure 4, a kind of preparation of anaerobe culture medium, inoculation and culture apparatus, including：Gas bomb group
1st, float gas flowmeter group 4, quartz glass tube 11 and aeration component, the gas bomb group 1 are three and are respectively provided with N2、CO2
And H2/CO2Gas bomb（1.1、1.2、1.3）；
The float gas flowmeter group 4 is three float gas flowmeters（4.1、4.2、4.3）One end is divided by stainless steel tube 3
Not with three gas bombs（1.1、1.2、1.3）It is corresponding to be connected, the first float gas flowmeter 4.1 and the second float gas flow
The other end of meter 4.2 is connected with the first cutting sleeve type three-way joint 5, then the port of export and the 3rd of the first cutting sleeve type three-way joint 5
The other end of float gas flowmeter 4.3 is connected with bite type tee ball valve 6, and the port of export of bite type tee ball valve 6 passes through outer
The stainless steel tube for being cased with rubber tube 7 is connected with the bottom of quartz glass tube 11；
Quartz glass tube 11 simultaneously has metal built with copper wire 10, the outside of quartz glass tube 11 winding glass fibre heating wire 9
Protective case, glass fibre heating wire connect single-phase contact type voltage regulator 19；Culture medium can be achieved and cultivate the deoxidation of atmosphere, solve
In culture medium and culture atmosphere containing oxygen cause anaerobe can not normal growth situation；
The top of quartz glass tube 11 is connected by being cased with the stainless steel tube of rubber tube 7 outside with the second cutting sleeve type three-way joint 5.1,
The one outlet end of second cutting sleeve type three-way joint 5.1 is sequentially connected with integrated valve hatpin valve 12, rubber tube 14, glass syringe
17th, stainless steel syringe needle 18；Another port of export of second cutting sleeve type three-way joint 5.1 is connected with integrated valve hatpin valve 12, then
Multiple aeration components are parallel with, are connected between aeration component with the 3rd cutting sleeve type three-way joint 5.2.
Gas bomb outlet is provided with steel cylinder pressure-reducing valve 2.
Stainless steel tube 3 is the stainless steel tube of 1/8 inch of external diameter.
Aeration component is made up of toggle switch valve 13, rubber tube 14, stainless steel syringe needle 15, anaerobism bottle 16.The side of being so designed that
Just the processing that exchanges for and carry out disinfection of syringe needle, and can insert in culture vessel.
The float gas flowmeter group 4, cutting sleeve type three-way joint（5、5.1、5.2）, bite type tee ball valve 6 and overall
Formula bonnet needle-valve 12 is installed in same stainless steel surface.
Two integrated valve hatpin valves 12 realize the preparation of anaerobic culture medium and the inoculation of anaerobe both can be same
Shi Jinhang can also be individually operated.
What apparatus of the present invention were realized in：
1st, deoxygenation system regenerates：After anaerobe inoculation and aerator use a period of time, the purple in quartz glass tube
Copper wire 10 can be oxidized blackening, and it need to be reduced.Single-phase contact type voltage regulator 19 is switched on power, connects and hydrogen is housed
The connected float gas flowmeter 1.3 of gas bomb, close two other float gas flowmeter, rotate bite type three-way ball
Valve 6 switches gas to hydrogen, opens integrated valve hatpin valve 12, treats that copper wire 10 is changed into aubergine from black in pipe, illustrates purple
Copper wire 10 is reduced, and deoxygenation system regeneration is completed.
2nd, prepared by anaerobic culture medium：Opening is equipped with the pressure-reducing valve of nitrogen gas steel cylinder 1.1 and equipped with carbon dioxide
The pressure-reducing valve of gas bomb 1.2 and its connected float gas flowmeter 4.1,4.2, close another float gas flowmeter
4.3, then bite type tee ball valve 6 and integrated valve hatpin valve 12 are opened, bite type tee ball valve 6 plays the work of converted gas
With, and open toggle switch valve 13, under be terminated with 14G stainless steels syringe needle 15, and stainless steel syringe needle 15 is inserted in anaerobism bottle 16
Carry out Air Exposure.
3rd, the inoculation of anaerobe：Open the pressure-reducing valve of the gas bomb 1.1 equipped with nitrogen and its connected float gas
Flowmeter body 4.1, two other float gas flowmeter 4.2 and 4.3 is closed, then open bite type tee ball valve 6 and monoblock type
Bonnet needle-valve 12, bite type tee ball valve 6 play a part of converted gas, glass syringe 17,20G stainless steels syringe needle 18 and outer
The mm rubber tubes 14 of footpath 5 are referred to as anaerobe classification inoculation apparatus.Switch on power during inoculation, about 30 min are preheated, then according to bacterium
Kind fostering requirement twists bite type tee ball valve 6 and is changed to nitrogen, and opens the monoblock type bonnet being connected with the mm rubber tubes 14 of external diameter 5
Needle-valve 12, it is inoculated with using anaerobe classification inoculation apparatus.
Embodiment one
Anaerobic bacteriaGeobacter metallireducensCulture
1st, prepared by ironic citrate culture medium
As shown in Fig. 2 a, Fig. 2 b, ironic citrate is used（Ferric Citrate, FC）As electron acceptor culture anaerobeGeobacter metallireducens.The process for preparation of ironic citrate culture medium is：Ironic citrate is weighed with electronic balance
13.7 g are stand-by, take 600 mL deionized waters to be positioned on magnetic force heating stirrer in 1 L beakers, add a small amount of ironic citrate,
A small amount of state for adding, maintaining the agitating and heating when adding medicament, and in this process again after ironic citrate to be added is complete
In it is appropriate add mL of distilled water 200 or so, ironic citrate is preferably dissolved, after all ironic citrates are completely dissolved,
First pH to 6.0~6.5 is adjusted with 10 mol/L NaOH（It is low with 5 mol/L or 2 mol/L when pH is close to 5.0
Concentration NaOH is adjusted）, then with 0.5 mol/L NaOH it is transferred to 7.0 or so（6.8~7.2）, it is separately added into after solution cooling
NaH2PO4 ·H2O 0.60 g、NH4Cl 0.25 g、KCl 0.10 g、NaHCO3 2.50 g、1 mmol/L Na2SeO4Solution
1 mL, the mL of trace element solution 10（Trace element solution forms：1.5 g/L aminotriacetic acids, 3.0 g/L MgSO4·7H2O,
1.0 g/L NaCl, 0.13 g/L ZnCl2, 0.5 g/L MnSO4·H2O, 0.1 g/L FeSO4·7H2O, 0.1 g/L
CaCl2·2H2O, 0.1 g/L CoCl2·6H2O, 0.01 g/L H3BO3, 0.01 g/L CuSO4·5H2O, 0.01 g/L KAl
(SO4)2·12H2O, 0.024 g/L Na2WO4·2H2O, 0.025 g/L Na2MoO4·2H2O, 0.025 g/L NiCl2·
2H2O）, the mL of vitamin solution 10（Vitamin solution forms：0.002 g/L biotins, 0.005 g/L pantothenic acid, 0.0001 g/L
B-12,0.005 g/L p-aminobenzoic acid, 0.005 g/L lipoic acids, 0.005 g/L nicotinic acid, 0.005 g/L thiamines,
0.005 g/L vitamin B2s, 0.01 g/L vitamin B6s, 0.002 g/L folic acid）, 1000 mL are settled to, the citric acid prepared
Iron culture medium is dispensed into 150 mL Anaerobic culturel bottles, every bottle of 50 mL, is cultivated after the completion of packing with homemade anaerobe
System and device carries out Air Exposure to it, and opening is equipped with the pressure-reducing valve of nitrogen gas steel cylinder 1.1 and equipped with carbon dioxide
The pressure-reducing valve of gas bomb 1.2 and its connected float gas flowmeter 4.1,4.2, close another float gas flowmeter
4.3, then open bite type tee ball valve 6 and integrated valve hatpin valve 12, and open toggle switch valve 13, under be terminated with 14G not
Become rusty draw point first 15, and syringe needle is inserted in anaerobism bottle 16 is inner to be aerated with deoxygenation.A length of 20 min during every bottle of aeration, used in aeration
Gas is the gaseous mixture of carbon dioxide and nitrogen（CO2/N2 = 20/80）.121oC after ironic citrate culture medium deoxidation treatment is high
Pressure 30 min of sterilizing, it is stand-by to be cooled to room temperature.
2、Geobacter metallireducensInoculation and culture
Switch on power about 30 min of preheating during inoculation, and the anaerobism bottle bottleneck equipped with ironic citrate culture medium first is put in into alcolhol burner
On slightly scorch, then with asepsis injector add the mL of 2 mol/L sodium acetate solutions 0.5, then will extract strain after it is sterile
Open the pressure-reducing valve of the gas bomb equipped with nitrogen close to the glass syringe for connecing bacterium device and its be connected in syringe needle one end
Float gas flowmeter, close two other float gas flowmeter, then open bite type tee ball valve and monoblock type bonnet
Needle-valve, then twist bite type tee ball valve according to Spawn incubation requirement and be changed to nitrogen, and open and be connected with the mm rubber tubes of external diameter 5
Integrated valve hatpin valve, glass syringe syringe needle one end constantly overflows anaerobic gas, it is ensured that inoculation link is in anaerobism shape
State, then by strain（Geobacter metallireducens）Inject in culture medium, inoculum concentration is 1 mL, is placed after inoculation
Cultivated in 30oC constant incubators about one week.Fig. 2 a are not to be inoculated withGeobacter metallireducensIronic citrate
Culture medium, Fig. 2 b are inoculationGeobacter metallireducensCultivate the ironic citrate culture medium after one week, the energy from figure
Enough find out that culture medium color is changed into light yellow from rufous, illustrate the anaerobism created using anaerobe inoculation with culture apparatus
Environment meetsGeobacter metallireducensGrowth demand, the device can be used for strictly anaerobic microorganism inoculation
With culture.
Embodiment two
Anaerobic bacteriaGeobacter sulfurreducensCulture
1、NBF（NB medium with fumarate）The preparation of culture medium
Experiment is with the electroactive bacterium of NBF medium culture strictly anaerobicsGeobacter sulfurreducens.NBF culture mediums
Process for preparation is：It is stand-by that the g of fumaric acid 4.64 is weighed with electronic balance, and 800 mL deionized waters are taken in 1 L beakers, add rich horse
Acid, after fumaric acid to be added is completely dissolved, first adjust pH to 6.5~7.0 with 10 mol/L NaOH（When pH is close to 6.0
When, adjusted with 10 mol/L or 2mol/L low concentrations NaOH）, it is transferred to 7.0 or so（6.8~7.2）, divide after solution cooling
CaCl is not added2·2H2O 0.04 g 、MgSO4·7H2O 0.1 g 、KCl 0.10 g 、NaHCO3 1.8 g 、Na2CO3·
H2O 0.5g、1 m mol/L Na2SeO4The mL of solution 1.0, the mL of 100X NB salting liquids 10（NB salting liquids form：42 g/L
K2HPO4, 22 g/L KH2PO4, 20 g/L NH4Cl, 36 g/L NaCl, 38 g/L KCl）, the mL of NB trace element solutions 10
（NB trace element solutions form：2.14 g/L aminotriacetic acids, 0.1 g/L MnCl2·4H2O, 0.3 g/L MgSO4·
7H2O, 0.2 g/L ZnSO4·7H2O, 0.17 g/L CoCl2·6H2O, 0.005 g/L H3BO3, 0.03 g/L CuCl2·
2H2O, 0.005 g/L KAl (SO4)2·12H2O, 0.02 g/L Na2WO4·2H2O, 0.9 g/L Na2MoO4·2H2O, 0.11
g/L NiSO4·6H2O）, the mL of vitamin solution 15（Vitamin solution forms：0.002 g/L biotins, 0.005 g/L pantothenic acid,
0.0001 g/L B-12,0.005 g/L lipoic acids, 0.005 g/L nicotinic acid, 0.005 g/L thiamines, 0.005 g/L dimension lifes
Plain B2,0.002 g/L folic acid）, it is settled to 1000 mL.The NBF solution for preparing completion is dispensed to 150 mL Anaerobic culturel bottles
In, every bottle of 50 mL, Air Exposure is carried out to it with homemade anaerobe culture systems device after the completion of packing, open dress
There are the pressure-reducing valve of nitrogen gas steel cylinder 1.1 and the pressure-reducing valve of the gas bomb 1.2 equipped with carbon dioxide and its be connected floating
Sub- gas flowmeter 4.1,4.2, closes another float gas flowmeter 4.3, then opens bite type tee ball valve 6 and entirety
Formula bonnet needle-valve 12, and open toggle switch valve 13, under be terminated with 14G stainless steels syringe needle 15, and syringe needle is inserted in anaerobism bottle 16
In be aerated with deoxygenation.A length of 20 min during every bottle of aeration, aeration are gases used for carbon dioxide and the gaseous mixture of nitrogen
（CO2/N2 = 20/80）.121oC after NBF culture medium deoxidation treatments, the min of autoclaving 30, it is stand-by to be cooled to room temperature.
2、Geobacter sulfurreducensInoculation and culture
Switch on power about 30 min of preheating during inoculation, is first put in the anaerobism bottle bottleneck equipped with NBF culture mediums on alcolhol burner slightly
Scorch, then with asepsis injector add the mL of 2 mol/L sodium acetate solutions 0.5, then by extract strain after asepsis injector
The pressure-reducing valve of the gas bomb equipped with nitrogen and its connected float are opened close to the glass syringe for connecing bacterium device in syringe needle one end
Gas flowmeter, two other float gas flowmeter is closed, then open bite type tee ball valve and integrated valve hatpin valve, so
Bite type tee ball valve is twisted according to Spawn incubation requirement afterwards and be changed to nitrogen, and open the monoblock type being connected with the mm rubber tubes of external diameter 5
Anaerobic gas are constantly overflowed in bonnet needle-valve, glass syringe syringe needle one end, it is ensured that inoculation link is in anaerobic state, then will
In strain injection culture medium, inoculum concentration is 1 mL, is positioned in 30oC constant incubators and cultivates about one week after inoculation.Fig. 3 a are
It is not inoculated withGeobacter sulfurreducens The pictorial diagram of NBF culture mediums, Fig. 3 b are inoculationGeobacter sulfurreducensAnd the NBF culture mediums after cultivating one week, it can be seen that culture medium color is changed into pink from colourless from figure
Color, this shows that the anaerobic environment equally meets anaerobic bacteriaGeobacter sulfurreducensGrowth demand, strain growing way
Well.
Embodiment three
Geobacter metallireducensWithMethanosarsina barkeri 800 co-culture coupling metabolism
Ethanol methane phase
1、NBM（NB modi ed medium）The preparation of culture medium
Experiment is with NBM（Media NB modified）For minimal medium, and connect using the simple anaerobe in laboratory is made by oneself
Kind and culture apparatus cultureGeobacter metallireducensWithMethanosarsina barkeri 800 co-
Culture coupling metabolism ethanol methane phases, to examine the validity of the device.The process for preparation of NBM culture mediums：First in beaker
The mL of distilled water about 800 is added, pH to 7.0 is adjusted with NaOH, CaCl is separately added into after solution cooling2·2H2O 0.3 g、
MgSO4·7H2O 0.3 g、FeSO4·7H2O 17 mg、Na2CO3·H2O 0.5 g、NaHCO3 1.8 g, 100X NB salting liquids
10 ml（NB salting liquids form：42 g/L K2HPO4, 22 g/L KH2PO4, 20 g/L NH4Cl, 36 g/L NaCl, 38 g/L
KCl), the ml of NB trace element solutions 10（NB trace element solutions form：2.14 g/L aminotriacetic acids, 0.1 g/L
MnCl2·4H2O, 0.3 g/L MgSO4·7H2O, 0.2 g/L ZnSO4·7H2O, 0.17 g/L CoCl2·6H2O, 0.005
g/L H3BO3, 0.03 g/L CuCl2·2H2O, 0.005 g/L KAl (SO4)2·12H2O, 0.02 g/L Na2WO4·2H2O,
0.9 g/L Na2MoO4·2H2O, 0.11 g/L NiSO4·6H2O）, 1 mmol/L Na2SeO4 1 ml, is settled to 1000
ML, then NBM culture mediums are placed in triangular flask, boiled after being sealed with aluminium-foil paper, the culture medium boiled is placed in ice domain, it is cold
But to room temperature.The NBM solution for preparing completion is subjected to Air Exposure with homemade anaerobe culture systems device to it（Such as
Fig. 1）, open the pressure-reducing valve equipped with nitrogen gas steel cylinder 1.1 and gas bomb 1.2 equipped with carbon dioxide pressure-reducing valve and
Its connected float gas flowmeter 4.1,4.2, another float gas flowmeter 4.3 is closed, then open bite type threeway
Ball valve 6 and integrated valve hatpin valve 12, and open toggle switch valve 13, under be terminated with 14G stainless steels syringe needle 15, and syringe needle is inserted
It is aerated in anaerobism bottle 16, a length of 20 min during every bottle of aeration, the gases used mixing for carbon dioxide and nitrogen of aeration
Gas（CO2/N2 = 20/80）.121oC after NBM culture medium deoxidation treatments, the min of autoclaving 30, it is stand-by to be cooled to room temperature.
2、 Geobacter metallireducensWithMethanosarsina barkeri 800 co-culture's
Inoculation and culture
Switch on power about 30 min of preheating during inoculation, and the anaerobism bottle bottleneck equipped with NBM culture mediums is put on alcolhol burner and slightly processed
It is roasting, then by being inoculated with mcg vitamin solution of the gas circuit with 1 mL syringes, 0.50 mL filtration sterilizations of addition（Vitamin solution
Composition：0.002 g/L biotins, 0.005 g/L pantothenic acid, 0.0001 g/L B-12,0.005 g/L lipoic acids, 0.005 g/L
Nicotinic acid, 0.005 g/L thiamines, 0.005 g/L vitamin B2s, 0.002 g/L folic acid）, 0.5 mL reducing agents（100 mmol/L
L-Cysteine and 50 mmol/L Na2S mixed solutions）And 0.5 mL ethanol solutions（2 mol/L）.By the nothing of inoculation
Bacterium syringe needle one end is close to the glass syringe for connecing bacterium device, the pressure-reducing valve and its phase of gas bomb of the opening equipped with nitrogen
Float gas flowmeter even, closes two other float gas flowmeter, then open bite type tee ball valve and integrated valve
Hatpin valve, then twist bite type tee ball valve according to Spawn incubation requirement and be changed to nitrogen, and open and the mm rubber tube phases of external diameter 5
Anaerobic gas are constantly overflowed in integrated valve hatpin valve even, glass syringe syringe needle one end, it is ensured that inoculation link is in anaerobism
State, then willMethanosarsina barkeri 800 withGeobacter metallireducensCo-culture
Inject in NBM culture mediums（Inoculum concentration is 10%, V/V）, it is placed under 37oC and cultivates, and utilizes gas-chromatography periodic detection anaerobism bottle
Overhead concentration of methane gas.Geobacter metallireducensWithMethanosarsina barkeri 800 co-
The concentration changes with time of methane is as shown in Figure 4 in culture anaerobism bottles.As seen from the figure：The concentration of preceding 20 d methane is 0,20
Methane concentration steeply rises after d, and this explanation uses the multi-functional anaerobe inoculation in self-control laboratory and culture apparatus successfully
CultivateGeobacter metallireducensWithMethanosarsina barkeri 800 co-culture。Geobacter metallireducensWithMethanosarsina barkeri 800 co-culture are strictly anaerobic
Microorganism, and two kinds of microorganisms can grow in homemade device, this has absolutely proved the device in strictly anaerobic microorganism
Inoculation and culture when validity.
Claims (4)
1. a kind of preparation of anaerobe culture medium, inoculation and culture apparatus, it is characterised in that：Including gas bomb group（1）, it is floating
Sub- gas flowmeter group（4）, quartz glass tube（11）With aeration component, the gas bomb group（1）N is respectively provided with for three2、
CO2And H2/CO2Gas bomb（1.1、1.2、1.3）；
The float gas flowmeter group（4）For three float gas flowmeters（4.1、4.2、4.3）One end passes through stainless steel tube
（3）Respectively with three gas bombs（1.1、1.2、1.3）It is corresponding to be connected, the first float gas flowmeter（4.1）With the second float
Gas flowmeter（4.2）The other end and the first cutting sleeve type three-way joint（5）It is connected, then the first cutting sleeve type three-way joint（5）
The port of export and the 3rd float gas flowmeter（4.3）The other end and bite type tee ball valve（6）It is connected, bite type three-way ball
Valve（6）The port of export by being cased with rubber tube outside（7）Stainless steel tube and quartz glass tube（11）Bottom is connected；
Quartz glass tube（11）Built with copper wire（10）, quartz glass tube（11）Outside winding glass fibre heating wire（9）And
With metal protection protector, glass fibre heating wire connects single-phase contact type voltage regulator（19）；
Quartz glass tube（11）Top by being cased with rubber tube outside（7）Stainless steel tube and the second cutting sleeve type three-way joint
（5.1）It is connected, the second cutting sleeve type three-way joint（5.1）One outlet end be sequentially connected with integrated valve hatpin valve（12）, rubber
Pipe（14）, glass syringe（17）, stainless steel syringe needle（18）；Second cutting sleeve type three-way joint（5.1）Another port of export connect
There is integrated valve hatpin valve（12）, multiple aeration components are then parallel with, are aerated between component with the 3rd cutting sleeve type three-way joint
（5.2）It is connected.
2. device as claimed in claim 1, it is characterised in that：Gas bomb outlet is provided with steel cylinder pressure-reducing valve（2）.
3. device as claimed in claim 1, it is characterised in that：Component is aerated by toggle switch valve（13）, rubber tube（14）, no
Become rusty draw point head（15）, anaerobism bottle（16）Composition.
4. device as claimed in claim 1, it is characterised in that：The float gas flowmeter group（4）, cutting sleeve type three-way joint
（5、5.1、5.2）, bite type tee ball valve（6）With integrated valve hatpin valve（12）It is installed in same stainless steel surface.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108641944A (en) * | 2018-05-17 | 2018-10-12 | 华东理工大学 | A kind of CO2It is biologically converted into the device and method of methane |
| CN113151105A (en) * | 2021-05-17 | 2021-07-23 | 东莞理工学院 | Pure culture method of anaerobic microorganisms |
| CN114458948A (en) * | 2022-02-23 | 2022-05-10 | 中国水产科学研究院黄海水产研究所 | Algae culture gas supply system and gas supply method thereof |
| CN115232713A (en) * | 2022-09-02 | 2022-10-25 | 安徽大学 | Portable laboratory simple and easy device of oxygen in getting rid of culture medium |
| CN115851427A (en) * | 2023-02-21 | 2023-03-28 | 北京林业大学 | Device and method for culturing geobacillus with ultrahigh-conductivity biological nanowire |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204981150U (en) * | 2015-07-17 | 2016-01-20 | 杭州秀川科技有限公司 | Deaerator |
| CN107175242A (en) * | 2017-07-07 | 2017-09-19 | 湖南大学 | Anaerobic culturel bottle inflation vacuumizes purging system |
| CN207793227U (en) * | 2017-11-03 | 2018-08-31 | 内蒙古科技大学 | The preparation of anaerobe culture medium, inoculation and culture apparatus |
-
2017
- 2017-11-03 CN CN201711070730.9A patent/CN107603858A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204981150U (en) * | 2015-07-17 | 2016-01-20 | 杭州秀川科技有限公司 | Deaerator |
| CN107175242A (en) * | 2017-07-07 | 2017-09-19 | 湖南大学 | Anaerobic culturel bottle inflation vacuumizes purging system |
| CN207793227U (en) * | 2017-11-03 | 2018-08-31 | 内蒙古科技大学 | The preparation of anaerobe culture medium, inoculation and culture apparatus |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108641944A (en) * | 2018-05-17 | 2018-10-12 | 华东理工大学 | A kind of CO2It is biologically converted into the device and method of methane |
| CN108641944B (en) * | 2018-05-17 | 2022-06-03 | 华东理工大学 | CO (carbon monoxide)2Device and method for bioconversion into methane |
| CN113151105A (en) * | 2021-05-17 | 2021-07-23 | 东莞理工学院 | Pure culture method of anaerobic microorganisms |
| CN114458948A (en) * | 2022-02-23 | 2022-05-10 | 中国水产科学研究院黄海水产研究所 | Algae culture gas supply system and gas supply method thereof |
| CN114458948B (en) * | 2022-02-23 | 2022-12-09 | 中国水产科学研究院黄海水产研究所 | Algae culture gas supply system and gas supply method thereof |
| CN115232713A (en) * | 2022-09-02 | 2022-10-25 | 安徽大学 | Portable laboratory simple and easy device of oxygen in getting rid of culture medium |
| CN115851427A (en) * | 2023-02-21 | 2023-03-28 | 北京林业大学 | Device and method for culturing geobacillus with ultrahigh-conductivity biological nanowire |
| WO2024174506A1 (en) * | 2023-02-21 | 2024-08-29 | 北京林业大学 | Apparatus and method for culturing geobacter metallireducens growing with ultra-high conductivity biological nanowires |
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