Cell-based assays using calcein acetoxymethyl ester show variation in fluorescence with treatment conditions

Authors

  • Fayth Miles University of California, Los Angeles, CA
  • Jill Lynch The University of Delaware
  • Robert Sikes Laboratory for Cancer Ontogeny and Therapeutics, Department of Biological Sciences The Center for Translational Cancer Research, The University of Delaware, Newark, DE

DOI:

https://doi.org/10.14440/jbm.2015.73

Keywords:

cellular fluorescence, calcein, cell-based assays, biochemical compounds, prostate cancer cells

Abstract

The use of fluorogenic compounds in cell and molecular biology has increased in both frequency and range of applications. However, such compounds may introduce artifacts in intracellular fluorescence and cell number estimations as a consequence of interaction with exogenous stimulants, necessitating the use of adequate controls for accurate measurements and valid conclusions. Using calcein acetoxymethyl ester (AM) in combination with various exogenous cellular treatments, we report that the standard practice of direct normalization of experimental values to controls is insufficient for fluorogenic measurements. Treatments applied to cells may influence intracellular conversion of the fluorogenic compound, thereby enhancing or decreasing fluorescence relative to controls. We hereby encourage caution and recommend normalization of cellular fluorescence within each treatment group before comparison to controls.

Author Biographies

Fayth Miles, University of California, Los Angeles, CA

Department of Epidemiology, Fielding School of Public Health, University of California, Los Angeles, CA

Senior Fellow Trainee

Jill Lynch, The University of Delaware

Laboratory for Cancer Ontogeny and Therapeutics, Department of Biological Sciences, The University of Delaware, Newark DE 19716

Graduate Student

Robert Sikes, Laboratory for Cancer Ontogeny and Therapeutics, Department of Biological Sciences The Center for Translational Cancer Research, The University of Delaware, Newark, DE

Department of Biological Sciences, associate professor; Center for Translational Cancer Research, director

University of Delaware, Newark, DE

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Published

2015-10-29

How to Cite

1.
Miles F, Lynch J, Sikes R. Cell-based assays using calcein acetoxymethyl ester show variation in fluorescence with treatment conditions. J Biol Methods [Internet]. 2015Oct.29 [cited 2022Aug.11];2(3):e29. Available from: https://jbmethods.org/jbm/article/view/73

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