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

Authors :
George SS
Steele CR
Ricci AJ
Source :
STAR protocols [STAR Protoc] 2021 Jun 30; Vol. 2 (3), pp. 100637. Date of Electronic Publication: 2021 Jun 30 (Print Publication: 2021).
Publication Year :
2021

Abstract

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).<br />Competing Interests: The authors declare no competing interests.<br /> (© 2021 The Authors.)

Details

Language :
English
ISSN :
2666-1667
Volume :
2
Issue :
3
Database :
MEDLINE
Journal :
STAR protocols
Publication Type :
Academic Journal
Accession number :
34258597
Full Text :
https://doi.org/10.1016/j.xpro.2021.100637