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Journal of Biomedical Optics • Open Access

Correlating two-photon excited fluorescence imaging of breast cancer cellular redox state with seahorse flux analysis of normalized cellular oxygen consumption
Author(s): Jue Hou; Heather J. Wright; Nicole Sy-Kay Chan; Richard D. H. Tran; Olga V. Razorenova; Eric O. Potma; Bruce J. Tromberg

Paper Abstract

Two-photon excited fluorescence (TPEF) imaging of the cellular cofactors nicotinamide adenine dinucleotide and oxidized flavin adenine dinucleotide is widely used to measure cellular metabolism, both in normal and pathological cells and tissues. When dual-wavelength excitation is used, ratiometric TPEF imaging of the intrinsic cofactor fluorescence provides a metabolic index of cells—the “optical redox ratio” (ORR). With increased interest in understanding and controlling cellular metabolism in cancer, there is a need to evaluate the performance of ORR in malignant cells. We compare TPEF metabolic imaging with seahorse flux analysis of cellular oxygen consumption in two different breast cancer cell lines (MCF-7 and MDA-MB-231). We monitor metabolic index in living cells under both normal culture conditions and, for MCF-7, in response to cell respiration inhibitors and uncouplers. We observe a significant correlation between the TPEF-derived ORR and the flux analyzer measurements (R=0.7901, p<0.001). Our results confirm that the ORR is a valid dynamic index of cell metabolism under a range of oxygen consumption conditions relevant for cancer imaging.

Paper Details

Date Published: 14 June 2016
PDF: 3 pages
J. Biomed. Opt. 21(6) 060503 doi: 10.1117/1.JBO.21.6.060503
Published in: Journal of Biomedical Optics Volume 21, Issue 6
Show Author Affiliations
Jue Hou, Beckman Laser Institute and Medical Clinic (United States)
Heather J. Wright, Univ. of California, Irvine (United States)
Nicole Sy-Kay Chan, Beckman Laser Institute and Medical Clinic (United States)
Richard D. H. Tran, Beckman Laser Institute and Medical Clinic (United States)
Olga V. Razorenova, Univ. of California, Irvine (United States)
Eric O. Potma, Univ. of California, Irvine (United States)
Bruce J. Tromberg, Beckman Laser Institute and Medical Clinic (United States)


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