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Anaerobic chamber

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The Anaerobic Chamber is a specialized laboratory equipment designed to maintain an oxygen-free environment. It creates a controlled atmosphere for the cultivation and examination of anaerobic microorganisms, which require the absence of oxygen to thrive.

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Lab products found in correlation

Sodium chloride (nacl), by Carl Roth (1 mentions) Proteose peptone, by Merck Group (1 mentions) Anaerobic atmosphere generation bags, by Merck Group (1 mentions) Horse serum, by Thermo Fisher Scientific (1 mentions) Yeast extract, by Thermo Fisher Scientific (1 mentions) α d glucose, by Merck Group (1 mentions) Hepes, by Merck Group (1 mentions) Potassium tetrathionate, by Merck Group (1 mentions) Streptomycin sulfate, by Merck Group (1 mentions) Gaspak system, by BD (1 mentions) X gal, by BD (1 mentions)

3 protocols using anaerobic chamber

1

Gardnerella Isolate Cultivation Protocol

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Gardnerella isolates of different species (52 (link)) were obtained from the Laboratory of Bacteriology, University of Ghent, Belgium. These isolates include strains purchased from culture collections and fresh isolates from BV patients obtained from the University Clinic Bruges. Gardnerella isolates were grown on chocolate (Choc) agar plates (Becton, Dickinson) under anaerobic conditions in an anaerobic chamber equipped with anaerobic atmosphere generation bags (Sigma-Aldrich) for 48 h. All isolates were cultured in New York City broth III (NYCB), consisting of 10 mM HEPES (Sigma-Aldrich), 15 g/L proteose peptone (Sigma-Aldrich), 3.8 g/L yeast extract (Thermo Fisher Scientific), 86 mM sodium chloride (Carl Roth), and 28 mM α-d-glucose (Sigma-Aldrich), supplemented with 10% horse serum (HS) (Thermo Fisher Scientific). Table S1 lists all Gardnerella and Lactobacillus isolates studied.
Landlinger C., Oberbauer V., Podpera Tisakova L., Schwebs T., Berdaguer R., Van Simaey L., Vaneechoutte M, & Corsini L. (2022). Preclinical Data on the Gardnerella-Specific Endolysin PM-477 Indicate Its Potential to Improve the Treatment of Bacterial Vaginosis through Enhanced Biofilm Removal and Avoidance of Resistance. Antimicrobial Agents and Chemotherapy, 66(5), e02319-21.
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2

Tetrathionate-Induced Bacterial Memory

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Tetrathionate-based memory was induced under in vitro conditions in either liquid culture or during growth on plates. For liquid culture induction, strains were backdiluted 1:1000 from overnight culture in SOC broth into pre-reduced anaerobic SOC broth and grown at 37°C in an anaerobic chamber (Coy Lab Products) using 7%H2/ 20% CO2/63% N2 culture gas. After 1h, potassium tetrathionate (Sigma) dissolved in pre-reduced SOC media was added to cultures and induction was undertaken at 37°C in the anaerobic chamber for 4h. Memory was assayed by plating and growth in aerobic conditions of serial dilutions of the bacteria on luria-broth (LB) + 300μg/mL streptomycin sulfate (Sigma) + 60μg/mL 5-Bromo-4-chloro-3-indolyl b-D-galactopyranoside (x-gal) (Santa Cruz Biotechnology) agar plates. Switching levels were estimated through counting greater than 250 blue and white colonies unless otherwise noted. In vitro testing on solid plates involved plating on LB + 300μg/mL streptomycin sulfate (Sigma) + 60μg/mL x-gal (Santa Cruz Biosciences) + 10 mM sodium tetrathionate (Sigma) agar. Growth was maintained in anaerobic conditions for approximately 8–12 hours using the Gaspak system (BD Biosciences), followed by further growth in aerobic conditions to allow development of memory and the x-gal reporter.
Riglar D.T., Giessen T.W., Baym M., Kerns S.J., Niederhuber M.J., Bronson R.T., Kotula J.W., Gerber G.K., Way J.C, & Silver P.A. (2017). Engineered bacteria function in the mammalian gut as long term live diagnostics of inflammation. Nature biotechnology, 35(7), 653-658.
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3

Microglial-Neuron Co-culture Ischemia

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In this experiment, purified microglial cells were seeded into primary neurons at DIV 5 with the cell density of 5 × 104 cells/well in a 24-well plate. Three hours later, co-cultures suffered OGD to mimic ischemia in vitro. Briefly, culture medium was replaced by serum and glucose-free DMEM, and co-cultures were placed in an anaerobic chamber (Sigma, USA) filled with 93% N2, 5% CO2 and 2% O2 for 1 h. Then co-cultures returned back into a normoxic chamber and cultured in normal culture medium. The cells were collected after 24 h to provide support for subsequent experiments.
Wei H.X., Guan Y.N., Chen P.P., Rao Z.Z, & Yang J.S. (2023). Upregulation of EphA4 deteriorate brain damage by shifting microglia M1-polarization via NF-κB signaling after focal cerebral ischemia in rats. Heliyon, 9(7), e18429.
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