Measurement of intracellular oxygen levels using fluorescence lifetime imaging microscopy (FLIM)

Dhruv Sud; Wei Zhong; David Beer; Mary-Ann Mycek

doi:10.1117/12.632827

7 October 2005 Measurement of intracellular oxygen levels using fluorescence lifetime imaging microscopy (FLIM)

Dhruv Sud, Wei Zhong, David Beer, Mary-Ann Mycek

Proceedings Volume 5859, Photon Migration and Diffuse-Light Imaging II; 585907 (2005) https://doi.org/10.1117/12.632827
Event: European Conferences on Biomedical Optics 2005, 2005, Munich, Germany

Abstract

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.

Citation Download Citation

Dhruv Sud, Wei Zhong, David Beer, and Mary-Ann Mycek "Measurement of intracellular oxygen levels using fluorescence lifetime imaging microscopy (FLIM)", Proc. SPIE 5859, Photon Migration and Diffuse-Light Imaging II, 585907 (7 October 2005); https://doi.org/10.1117/12.632827

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