Potentiometric dyes: new modalities for optical imaging of membrane potential

Leslie M. Loew

doi:10.1117/12.239555

10 May 1996 Potentiometric dyes: new modalities for optical imaging of membrane potential

Leslie M. Loew

Proceedings Volume 2678, Optical Diagnostics of Living Cells and Biofluids; (1996) https://doi.org/10.1117/12.239555
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

Voltage sensitive dyes permit the measurement of biomembrane electrical activity where microelectrode measurements are unsuitable or inadequate. This laboratory has developed 2 class of such dyes as well as some new optical methods for mapping membrane potential in single cells. One class of compounds consists of amphiphilic membrane staining dyes containing a putative electrochromic chromophore aligned perpendicular to the membrane/aqueous interface. We have used these dyes to map the membrane potential along the surface of single cells with high resolution dual wavelength ratiometric imaging microscopy techniques. We have found intrinsic regional variations in the electrical properties of cell membranes. Adding chirality produces compounds that display remarkable efficiencies for second harmonic generation. Further, the second harmonic signal can be generated from a dye-stained membrane and is modulatable by changing the membrane voltage. This can form the basis for measurements of membrane electrophysiology at high 3D resolution with infrared light. A second class of dyes are membrane-permeant cations which act as Nernstian indicators. We have developed calibration methods which allow the use of these compounds to quantitate the membrane potential of individual mitochondria from confocal or widefield images of living cells.

Citation Download Citation

Leslie M. Loew "Potentiometric dyes: new modalities for optical imaging of membrane potential", Proc. SPIE 2678, Optical Diagnostics of Living Cells and Biofluids, (10 May 1996); https://doi.org/10.1117/12.239555

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