Measurement of intracellular oxygen levels using Fluorescence Lifetime Imaging Microscopy (FLIM)

摘要

Intracellular oxygen levels were measured in vivo under physiological-temperature controlled conditions by monitoring the fluorescence lifetime of the oxygen sensitive dye ruthenium tris(2,2'-dipyridyl) dichloride hexahydrate (RTDP). We employed fluorescence lifetime imaging microscopy (FLIM) and an independent oxygen sensor to calibrate changes in RTDP lifetime with corresponding changes in oxygen level. The FLIM method reproducibly quantified oxygen levels in living human cells cultured from a (Barrett's) adenocarcinoma columnar cell line (SEG-1). Approaches such as those developed here should prove useful for studying oxygen gradients in 3-D biological specimens and for monitoring cellular metabolic status.