Krycer James R, Diskin Ciana, Nelson Marin E, Zeng Xiao-Yi, Fazakerley Daniel J, James David E
School of Life and Environmental Sciences, Charles Perkins Centre, The University of Sydney.
Sydney Medical School, The University of Sydney.
Biotechniques. 2018 Jan 1;64(1):27-29. doi: 10.2144/000114629.
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.
细胞代谢研究已变得更加高通量,典型的细胞培养实验在多孔板(微孔板)中进行。当试图收集气态产物,如二氧化碳(CO₂)时,这种形式带来了挑战,因为这需要一个密封环境以及一个与生物样品分开的容器。为解决这一限制,我们开发了一种气体捕获方案,在密封的细胞培养多孔板中使用穿孔塑料盖。我们利用这种捕获设计来测量葡萄糖和脂肪酸代谢产生的CO₂,以及半胱氨酸处理的细胞产生的硫化氢。我们的数据清楚地表明,这种气体捕获器可应用于液体和固体气体收集介质,可用于研究贴壁细胞和悬浮细胞产生气态产物的情况。由于我们的气体捕获器可适配各种尺寸的多孔板,它们提供了一种方便、经济高效的解决方案,能够适应代谢研究中高通量测量的趋势。