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29 January 2015 High-throughput measurements of the optical redox ratio using a commercial microplate reader
Taylor M. Cannon, Amy T. Shah, Alex J. Walsh, Melissa C. Skala
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Journal of Biomedical Optics, Vol. 20, Issue 1, 010503 (January 2015). https://doi.org/10.1117/1.JBO.20.1.010503
Abstract
There is a need for accurate, high-throughput, functional measures to gauge the efficacy of potential drugs in living cells. As an early marker of drug response in cells, cellular metabolism provides an attractive platform for high-throughput drug testing. Optical techniques can noninvasively monitor NADH and FAD, two autofluorescent metabolic coenzymes. The autofluorescent redox ratio, defined as the autofluorescence intensity of NADH divided by that of FAD, quantifies relative rates of cellular glycolysis and oxidative phosphorylation. However, current microscopy methods for redox ratio quantification are time-intensive and low-throughput, limiting their practicality in drug screening. Alternatively, high-throughput commercial microplate readers quickly measure fluorescence intensities for hundreds of wells. This study found that a commercial microplate reader can differentiate the receptor status of breast cancer cell lines (p<0.05) based on redox ratio measurements without extrinsic contrast agents. Furthermore, microplate reader redox ratio measurements resolve response (p<0.05) and lack of response (p>0.05) in cell lines that are responsive and nonresponsive, respectively, to the breast cancer drug trastuzumab. These studies indicate that the microplate readers can be used to measure the redox ratio in a high-throughput manner and are sensitive enough to detect differences in cellular metabolism that are consistent with microscopy results.
CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
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Taylor M. Cannon, Amy T. Shah, Alex J. Walsh, and Melissa C. Skala "High-throughput measurements of the optical redox ratio using a commercial microplate reader," Journal of Biomedical Optics 20(1), 010503 (29 January 2015). https://doi.org/10.1117/1.JBO.20.1.010503
Published: 29 January 2015
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Cited by 21 scholarly publications.
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KEYWORDS
Luminescence
Breast cancer
Mode conditioning cables
Microscopy
Receptors
Optical testing
Drug development
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