Fluorescent pH probes, fluorescent proteins, and intrinsic cellular fluorochromes are tools to study cytosolic pH (pHcyt) in mammalian cells.

Gloria M. Martinez; Lauren S. Gollahon; Keri Shafer; Sowmini K. Oomman; Christian Busch; Raul Martinez-Zaguilan

doi:10.1117/12.432492

2 July 2001 Fluorescent pH probes, fluorescent proteins, and intrinsic cellular fluorochromes are tools to study cytosolic pH (pH^cyt) in mammalian cells.

Gloria M. Martinez, Lauren S. Gollahon, Keri Shafer, Sowmini K. Oomman, Christian Busch, Raul Martinez-Zaguilan

Author Affiliations +

Proceedings Volume 4259, Biomarkers and Biological Spectral Imaging; (2001) https://doi.org/10.1117/12.432492
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

Our understanding of intracellular pH homeostatis in eukaryotic systems has been enhanced since the introduction of carboxyfluorescein diacetate as a useful pH probe more than 20 years ago. BCECF, a derivative of this earlier fluoroprobe has dominated the field. In the past 10 years, SNARF-1 has emerged as an alternative pH probe. Recently, a novel derivative of BCECF, BCPCF has been developed. Green Fluorescent Proteins (GFPs) have also been used recently to monitor pH in a non invasive manner in several cell types. Here, we report that human mammary epithelial cells can be transfected with the gene encoding for cyan (CFP), green (GFP), and yellow (YFP), to study cytosolic pH. The novel red fluorescent protein (DsRed) is not sensitive to pH. Multidrug resistance (MDR) has been associated with altered cytosolic pH homeostasis. We show that experimental maneuvers that decrease pHⁱⁿ enhance the efficacy of chemotherapeutic drugs. We also show that short pulses of UV-B light elicited acidosis in cells, as evaluated by ratio ion cell imaging, and confocal/spectral imaging microscopy. During the course of these experiments we noticed that cells exhibit intrinsic fluorochromes that can be used to monitor pH in living cells.

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Gloria M. Martinez, Lauren S. Gollahon, Keri Shafer, Sowmini K. Oomman, Christian Busch, and Raul Martinez-Zaguilan "Fluorescent pH probes, fluorescent proteins, and intrinsic cellular fluorochromes are tools to study cytosolic pH (pH^cyt) in mammalian cells.", Proc. SPIE 4259, Biomarkers and Biological Spectral Imaging, (2 July 2001); https://doi.org/10.1117/12.432492

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