Proliferation, colony formation assay, and single cell gel electrophoresis (a.k.a. Comet) assays, also known as the Comet assay were performed as previously described (17 (link)). Hypoxia was created by placing cells into Heracell™ 150i Tri-Gas Incubator (Thermo Scientific) with 1% oxygen, 94% nitrogen, 5% CO2. Cells were exposed to a 137Cs γ-ray source (Atomic Energy of Canada) at the specified doses.
Heracell 150i tri gas incubator
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany
The Heracell™ 150i Tri-Gas Incubator is a laboratory equipment designed to provide a controlled environment for cell and tissue culture applications. It features the ability to regulate temperature, humidity, and gas composition (CO2, O2, and N2) within the incubation chamber.
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Lab products found in correlation
4 protocols using heracell 150i tri gas incubator
1
Radiation Effect Assays
2
Culturing and Characterizing Campylobacter jejuni
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Strains of C. jejuni used in this study (Table 1
For strain construction, motility agar assays and negative-stain electron microscopy (performed at Imperial College London), cultures were grown in a Heracell 150i trigas incubator (Thermo-Fischer) set to 5% O2, 10% CO2, 85% N2 at 37° C.
For fluorescence microscopy experiments (performed at Gakushuin University), cultures were grown at 37° C using Mitsubishi Anaeropack MicroAero gas generator packs.
For strain construction, motility agar assays and negative-stain electron microscopy (performed at Imperial College London), cultures were grown in a Heracell 150i trigas incubator (Thermo-Fischer) set to 5% O2, 10% CO2, 85% N2 at 37° C.
For fluorescence microscopy experiments (performed at Gakushuin University), cultures were grown at 37° C using Mitsubishi Anaeropack MicroAero gas generator packs.
3
Cell Proliferation and Clonogenicity Assays
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To assay for proliferation, 3000 cells were plated onto 24-well plates and maintained in media overnight. A colorimetric MTT assay was used to assess cell number by optical density after 3 days as described previously (Yoon et al, 1999 (link)). Data reflect the mean of six samples.
To assay for colony forming ability, 200–500 cells were plated onto 60 mm culture dishes and incubated for 7–14 days. Colonies were stained with 0.5% crystal violet in 6% glutaraldehyde solution. The number of colonies consisting of 50 or more cells was scored. Data reflect the mean of nine samples.
Hypoxia was created by placing cells into Heracell 150i Tri-Gas Incubator (Thermo Scientific, Waltham, MA, USA) with 1% oxygen, 94% nitrogen, and 5% CO2. Each experiment was performed at least three times.
To assay for colony forming ability, 200–500 cells were plated onto 60 mm culture dishes and incubated for 7–14 days. Colonies were stained with 0.5% crystal violet in 6% glutaraldehyde solution. The number of colonies consisting of 50 or more cells was scored. Data reflect the mean of nine samples.
Hypoxia was created by placing cells into Heracell 150i Tri-Gas Incubator (Thermo Scientific, Waltham, MA, USA) with 1% oxygen, 94% nitrogen, and 5% CO2. Each experiment was performed at least three times.
4
Culturing P. falciparum for Mosquito Infections
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P. falciparum NF54 parasites (a kind gift of R. Sauerwein) were cultured in O+ human red blood cells (Haema, Berlin) at 37 °C, 4% CO2 and 3% O2 in a Heracell 150i Tri-gas incubator (Thermo Fisher Scientific). For gametocyte production, asynchronous asexual parasite cultures were diluted to 1% parasitemia and 4% hematocrit and maintained for 15–16 d with daily change of RPMI-1640 medium (Thermo Fisher Scientific) supplemented with 10% human A+ serum (Haema, Berlin) and 10 mM hypoxanthine (c-c-Pro) until mosquito infections.
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