A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells

Shefin S. George; Charles R. Steele; Anthony J. Ricci

STAR Protocols (Sep 2021)

A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells

Shefin S. George,
Charles R. Steele,
Anthony J. Ricci

Affiliations

Shefin S. George: Department of Otolaryngology-Head and Neck Surgery, School of Medicine, 240 Pasteur Drive, Stanford University, Stanford, CA 94305, USA; Corresponding author
Charles R. Steele: Department of Mechanical Engineering, Building 520, 440 Escondido Mall, Stanford University, Stanford, CA 94305, USA
Anthony J. Ricci: Department of Otolaryngology-Head and Neck Surgery, School of Medicine, 240 Pasteur Drive, Stanford University, Stanford, CA 94305, USA; Department of Molecular and Cellular Physiology, School of Medicine, 300 Pasteur Drive, Stanford University, Stanford, CA 94305, USA; Corresponding author

Journal volume & issue: Vol. 2, no. 3
p. 100637

Abstract

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Summary: Fluorescence recovery after photobleaching (FRAP) has been widely used to monitor membrane properties by measuring the lateral diffusion of fluorescent particles. This protocol describes how to perform two-photon FRAP on the stereocilia of live cochlear inner hair cells using a lipophilic dye, di-3-ANEPPDHQ, to assess the stereociliary membrane diffusivity. We also detail two-photon FRAP microscope setup and calibration, as well as FRAP parameter setting and data analysis.For complete details on the use and execution of this protocol, please refer to George et al. (2020).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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