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Vinyl anaerobic chamber

Manufactured by Coy Laboratory Products
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The Vinyl Anaerobic Chamber is a sealed enclosure designed to maintain an anaerobic environment for various laboratory applications. It provides a controlled atmosphere with low oxygen levels to support the growth and study of anaerobic organisms.

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Lab products found in correlation

Powerwave 340, by Agilent Technologies (2 mentions) Eon microplate spectrophotometer, by Agilent Technologies (1 mentions) Beadblaster 24, by Benchmark Scientific (1 mentions) Gifu anaerobic medium, by Nissui Pharmaceutical (1 mentions) Anaeropack, by Mitsubishi (1 mentions) Emstat3, by PalmSens (1 mentions) Milli q, by Merck Group (1 mentions) Bhi broth, by BD (1 mentions) P5958, by Merck Group (1 mentions) S318 100, by Thermo Fisher Scientific (1 mentions) D16 500, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Anaerobic chamber (228 citations) Coy lab vinyl anaerobic chamber (4 citations) Type A vinyl anaerobic chamber (2 citations) Vinyl anaerobic airlock chamber (2 citations)

17 protocols using vinyl anaerobic chamber

1

Anaerobic Cultivation and Cell Harvesting

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Cultures were prepared in a vinyl anaerobic chamber (Coy Laboratory Products). Media, as described above, were dispensed in serum vials (50 ml of medium per 110-ml glass bottle) and inoculated with 250 μl from a stock culture of Dm RS-1 and Dh AM13 and incubated for ~1 month at 30°C and 35°C, respectively. Cell pellets were recovered from Dh AM13 cultures and from Dm RS-1 cultures by centrifugation at 8,000 g for 20 min. Cell pellets were then resuspended in anoxic sterile water for carbon quantification in the elemental analyzer (see below).
Nabeh N., Brokaw C, & Picard A. (2022). Quantification of Organic Carbon Sequestered by Biogenic Iron Sulfide Minerals in Long-Term Anoxic Laboratory Incubations. Frontiers in Microbiology, 13, 662219.
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2

Bifidobacterium carbon source utilization

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Bifidobacterium bifidum strain RJX-1201, Bifidobacterium breve RJX-1202 and Bifidobacterium longum RJX-1203 were streaked on brain heart infusion agar (BD) supplemented with 1% vitamin K/hemin solution (BD; sBHI), and incubated for 48 h in a vinyl anaerobic chamber (Coy Laboratory Products) containing 5% CO2, 5% H2 and 90% N2 and maintained at 37 °C. Cells were transferred to sBHI liquid medium (BHI broth, BD, supplemented as above) and grown for 24 h in anaerobic conditions. Cultures were washed twice with PBS and optical density at 600 nm (OD600) was measured using a BioTek PowerWave 340 plate reader. OD600 was normalized to 0.2 for all strains and 5 μl bacteria inoculum was added to a final volume of 200 μl containing 10% sBHI and 125 mM carbon source (glucose, fucose, galactose or lactose) in a 96-well plate. OD600 was measured in the plate reader every hour for 48 h with 5 s of medium shaking before each measurement. All of the measurements were normalized to a medium-only blank. Experiment was repeated three times (n = 3) in triplicate and one representative experiment is shown. Error bars are s.d. of three technical replicates.
Vatanen T., Franzosa E.A., Schwager R., Tripathi S., Arthur T.D., Vehik K., Lernmark Å., Hagopian W.A., Rewers M.J., She J.X., Toppari J., Ziegler A.G., Akolkar B., Krischer J.P., Stewart C.J., Ajami N.J., Petrosino J.F., Gevers D., Lähdesmäki H., Vlamakis H., Huttenhower C, & Xavier R.J. (2018). The human gut microbiome in early-onset type 1 diabetes from the TEDDY study. Nature, 562(7728), 589-594.
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3

Anaerobic Purification of T. maritima Enzymes

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T. maritima cells (1 g wet weight) were resuspended with 5 mL of anaerobic 50 mM phosphate buffer (pH 7.4), and 20 mg lysozyme was added. After incubating at 37°C for 6 h, the cells were disrupted by ultrasonication in a vinyl anaerobic chamber (Coy Laboratory Products, Inc.) filled with 95% N2 and 5% H2. After centrifugation at 10,000 × g to remove cell debris, the cell extracts were centrifuged at 100,000 × g, and the supernatant and the membrane fraction were used for enzyme assays.
Liang J., Huang H., Wang Y., Li L., Yi J, & Wang S. (2022). A Cytoplasmic NAD(P)H-Dependent Polysulfide Reductase with Thiosulfate Reductase Activity from the Hyperthermophilic Bacterium Thermotoga maritima. Microbiology Spectrum, 10(4), e00436-22.
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4

Anaerobic Purification of T. maritima Enzymes

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T. maritima cells (1 g wet weight) were resuspended with 5 mL of anaerobic 50 mM phosphate buffer (pH 7.4), and 20 mg lysozyme was added. After incubating at 37°C for 6 h, the cells were disrupted by ultrasonication in a vinyl anaerobic chamber (Coy Laboratory Products, Inc.) filled with 95% N2 and 5% H2. After centrifugation at 10,000 × g to remove cell debris, the cell extracts were centrifuged at 100,000 × g, and the supernatant and the membrane fraction were used for enzyme assays.
Liang J., Huang H., Wang Y., Li L., Yi J, & Wang S. (2022). A Cytoplasmic NAD(P)H-Dependent Polysulfide Reductase with Thiosulfate Reductase Activity from the Hyperthermophilic Bacterium Thermotoga maritima. Microbiology Spectrum, 10(4), e00436-22.
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5

Culturing Engineered Bacteroides Strains

Cited 1 time
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List of strains and expected fluorescence proteins are reported in Supplementary Table 1. We acquired strains from the Sonnenburg lab (Stanford University), who engineered reference Bacteroides strains (ATCC) by inserting chromosomally integrated fluorescence reporters (Whitaker et al., 2017 (link)). Frozen glycerol stocks of Bacteroides were thawed for ~30–60 min, streaked on Blood Agar Plates, then grown overnight for ~48 h at 37°C anaerobically (85% N2, 10% CO2, 5% H2) in a vinyl anaerobic chamber (Coy Laboratory Products). Strains were subcultured and passaged twice on agar using the same conditions before any of the experiments included in this manuscript.
Midani F.S, & David L.A. (2023). Tracking defined microbial communities by multicolor flow cytometry reveals tradeoffs between productivity and diversity. Frontiers in Microbiology, 13, 910390.
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6

Cultivation of C. thermocellum DSM 1313

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C. thermocellum DSM 1313 was obtained from the DSMZ microorganism collection (www.dsmz.de). All strains used in this study are listed in Table 2. Strains were stocked by addition of 25% (vol/vol) glycerol to overnight cultures grown in CTFUD (described by Olson and Lynd [38 (link)]) and stored in 1-ml aliquots in cryovials at −80°C. The strains were prepared in a vinyl anaerobic chamber from Coy Laboratory Products (COY Labs [Grass Lake, MI] or TG Instruments [Helsingborg, Sweden]) with 5% H2, 10% CO2, and 85% N2.
Yayo J., Kuil T., Olson D.G., Lynd L.R., Holwerda E.K, & van Maris A.J. (2021). Laboratory Evolution and Reverse Engineering of Clostridium thermocellum for Growth on Glucose and Fructose. Applied and Environmental Microbiology, 87(9), e03017-20.
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7

Anaerobic Characterization of DXP Synthase

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Unless otherwise noted, all reagents were obtained from commercial sources. All DNA primers were purchased from Integrated DNA Technologies. E. coli DXP synthase and E. coli MEP synthase (IspC) were overexpressed and purified as reported previously.23 (link),40 (link) Anaerobic spectropho-tometric analyses were performed on a Tecan infinite M nano UV/visible plate reader (Switzerland) that is housed inside a Coy Laboratory Products (Grass Lake, MI) vinyl anaerobic chamber. Anaerobic conditions for experiments that were conducted outside of the chamber (e.g., CD) were established in the anaerobic chamber, and anaerobic solutions were transferred to airtight, septum-capped vials or cuvettes. For steady state CD studies, spectra were recorded on an Aviv (Lakewood, NJ) 420 CD spectrometer. Trypsin digest peptides were sequenced by the Taplin Mass Spectrometry Facility at Harvard University (Cambridge, MA). Intact masses of trypsin digest peptides were determined by the Mass Spectrometry Core at The Johns Hopkins University School of Medicine. HDX-MS experiments were carried out on a 7T Bruker Daltonics Fourier Transform Mass Spectrometer.
DeColli A.A., Zhang X., Heflin K.L., Jordan F, & Freel Meyers C.L. (2019). Active Site Histidines Link Conformational Dynamics with Catalysis on Anti-Infective Target 1-Deoxy-d-xylulose 5-Phosphate Synthase. Biochemistry, 58(49), 4970-4982.
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8

Clostridium difficile Vegetative Cell Isolation

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Spore stocks of C. difficile strains 630 (ATCC BAA-1382) and VPI 10463 (ATCC 43255) were prepared as previously described (15 (link)).
Vegetative C. difficile was cultured and manipulated in a vinyl anaerobic chamber (Coy Laboratory Products) at 37°C. To generate the vegetative cell inoculum, strain VPI 10463 was streaked from the spore stock onto a plate of prereduced cycloserine-cefoxitin-fructose agar containing 0.1% taurocholate (TCCFA). TCCFA was prepared as previously described (15 (link)). The following day, a colony from the TCCFA was subcultured onto brain heart infusion (BHI) agar. After overnight growth, a colony from this plate was inoculated into 5 ml of brain heart infusion broth supplemented with 0.01% cysteine (BHIS) and incubated overnight. The following morning, broth-grown bacteria were harvested by centrifugation at 4,000 × g for 13 min. The supernatant was discarded, and the pellet was resuspended in 5 ml of sterile phosphate-buffered saline (PBS) (Gibco; catalog no. 10010023). The pellet was washed two more times before being used to challenge mice.
Leslie J.L., Jenior M.L., Vendrov K.C., Standke A.K., Barron M.R., O’Brien T.J., Unverdorben L., Thaprawat P., Bergin I.L., Schloss P.D, & Young V.B. (2021). Protection from Lethal Clostridioides difficile Infection via Intraspecies Competition for Cogerminant. mBio, 12(2), e00522-21.
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9

Bacterial Growth Under Aerobic and Anaerobic Conditions

Cited 2 times
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Bacterial strains were cultured overnight in LB broth either aerobically or in a vinyl anaerobic chamber (Coy Laboratory Products, Grass Lake, MI). On the following day, cultures were incubated in M9 (Life Technologies) medium with or without various carbon supplementation as described elsewhere (41 (link), 42 (link)). Strains were cultured for 24 to 196 h at 37°C either aerobically or in the anaerobic chamber, respectively. Absorbance readings at 600 nm were taken every 15 min using an Eon microplate spectrophotometer with Gen5 software (BioTek, Winooski, VT).
Martin R.M., Cao J., Wu W., Zhao L., Manthei D.M., Pirani A., Snitkin E., Malani P.N., Rao K, & Bachman M.A. (2018). Identification of Pathogenicity-Associated Loci in Klebsiella pneumoniae from Hospitalized Patients. mSystems, 3(3), e00015-18.
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10

Gnotobiotic Microbiome Transfer for Colorectal Cancer

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Wild-type and villin-TLR4 germ-free (GF) mice were generated at the University of Miami Gnotobiotic Facility. Microbial engraftment was performed by transferring the mucosa-associated microbiota from C57Bl/6J or villin-TLR4 donor mice to wild-type GF recipient mice. Briefly, the mucosa-associated microbiota was extracted by homogenizing flushed colons in Hank’s balanced salt solution using a BeadBlaster-24 (Benchmark) in a vinyl anaerobic chamber (Coy Laboratory Products). One mL of Hank’s balanced salt solution was used for every 50 mg of colon. Wild-type GF recipient mice were orally gavaged with 200 μL of the resulting slurry and housed in separate biocontainment unit isocages depending on the donor mouse microbiome (vilin-TLR4 versus C57Bl/6J). After three weeks of engraftment, 2 recipient mice per each donor mouse were euthanized and a piece of mucosa was analyzed by 16s rRNA sequencing to verify engraftment. The remaining mice underwent the azoxymethane / dextran sulfate sodium (AOM-DSS) model of tumorigenesis.
Burgueño J.F., Fritsch J., Gonzalez E.E., Landau K.S., Santander A.M., Fernández I., Hazime H., Davies J.M., Santaolalla R., Phillips M.C., Diaz S., Dheer R., Brito N., Pignac-Kobinger J., Fernández E., Conner G.E, & Abreu M.T. (2020). Epithelial TLR4 signaling activates DUOX2 to induce microbiota-driven tumorigenesis. Gastroenterology, 160(3), 797-808.e6.
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