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

STAR Protocols

Charles R. Steele¹  Anthony J. Ricci²  Shefin S. George³ 

[1] Corresponding author;Department of Mechanical Engineering, Building 520, 440 Escondido Mall, Stanford University, Stanford, CA 94305, USA;Department of Otolaryngology-Head and Neck Surgery, School of Medicine, 240 Pasteur Drive, Stanford University, Stanford, CA 94305, USA;

关键词: Cell Membrane;    Microscopy;    Neuroscience;    Molecular/Chemical Probes;   

DOI  :  

来源: DOAJ

【 摘 要 】

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).

【 授权许可】

Unknown