Measurement of cell respiration and oxygenation in standard multichannel biochips using phosphorescent O2-sensitive probes

Abstract

Measurement of cell oxygenation and oxygen consumption is useful for studies of cell bioenergetics, metabolism, mitochondrial function, drug toxicity and common pathophysiological conditions. Here we present a new platform for such applications which uses commercial multichannel biochips (μ-slides, Ibidi) and phosphorescent O₂ sensitive probes. This platform was evaluated with both extracellular and intracellular O₂ probes, several different cell types and treatments including mitochondrial uncoupling and inhibition, depletion of extracellular Ca²⁺ and inhibition of V-ATPase and histone deacetylases. The results show that compared to the standard microwell plates currently used, the μ-slide platform provides facile O₂ measurements with both suspension and adherent cells, higher sensitivity and reproducibility, and faster measurement time. It also allows re-perfusion and multiple treatments of cells and multi-parametric analyses in conjunction with other probes. Optical measurements are conducted on standard fluorescence readers and microscopes.