A gas trapping method for high-throughput metabolic experiments
Author(s) -
James R. Krycer,
Ciana Diskin,
Marin E. Nelson,
Zeng Xiao-Yi,
Daniel J. Fazakerley,
David E. James
Publication year - 2018
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114629
Subject(s) - carbon dioxide , throughput , trap (plumbing) , suspension (topology) , trapping , acid gas , chemistry , hydrogen sulfide , chromatography , materials science , process engineering , computer science , environmental science , sulfur , biology , organic chemistry , inorganic chemistry , engineering , telecommunications , ecology , mathematics , homotopy , environmental engineering , pure mathematics , wireless
Research into cellular metabolism has become more high-throughput, with typical cell-culture experiments being performed in multiwell plates (microplates). This format presents a challenge when trying to collect gaseous products, such as carbon dioxide (CO2), which requires a sealed environment and a vessel separate from the biological sample. To address this limitation, we developed a gas trapping protocol using perforated plastic lids in sealed cell-culture multiwell plates. We used this trap design to measure CO2 production from glucose and fatty acid metabolism, as well as hydrogen sulfide production from cysteine-treated cells. Our data clearly show that this gas trap can be applied to liquid and solid gas-collection media and can be used to study gaseous product generation by both adherent cells and cells in suspension. Since our gas traps can be adapted to multiwell plates of various sizes, they present a convenient, cost-effective solution that can accommodate the trend toward high-throughput measurements in metabolic research.
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