Application of fluorescence polarization microscopy to measure fluorophore orientation in the outer hair cell plasma membrane

Jennifer N. Greeson; Robert M. Raphael

doi:10.1117/1.2717499

1 March 2007 Application of fluorescence polarization microscopy to measure fluorophore orientation in the outer hair cell plasma membrane

Jennifer N. Greeson, Robert M. Raphael

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Journal of Biomedical Optics, Vol. 12, Issue 2, 021002 (March 2007). https://doi.org/10.1117/1.2717499

Abstract

The biophysical properties and organization of cell membranes regulate many membrane-based processes, including electromotility in outer hair cells (OHCs) of the cochlea. Studies of the membrane environment can be carried out by measuring the orientation of membrane-bound fluorophores using fluorescence polarization microscopy (FPM). Due to the cylindrical shape of OHCs, existing FPM theory developed for spherical cells is not applicable. We develop a new method for analyzing FPM data suitable for the quasi-cylindrical OHC. We present the theory for this model, as well as a study of the orientation of the fluorescent probe pyridinium, 4-[2-[6-(dioctylamino)-2-naphthalenyl]ethenyl]-1-(3-sulfopropyl) (di-8-ANEPPS) in the OHC membrane. Our results indicate that the absorption transition dipole moment of di-8-ANEPPS orients symmetrically about the membrane normal at 27 deg with respect to the plane of the membrane. The observed agreement between theoretical predictions and experimental measurements establishes the applicability of FPM to study OHC plasma membrane properties.

©(2007) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Jennifer N. Greeson and Robert M. Raphael "Application of fluorescence polarization microscopy to measure fluorophore orientation in the outer hair cell plasma membrane," Journal of Biomedical Optics 12(2), 021002 (1 March 2007). https://doi.org/10.1117/1.2717499

Published: 1 March 2007

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KEYWORDS

Molecules

Polarization

Absorption

Luminescence

Microscopy

Plasma

Polarizers

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