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

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-...

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Autores principales: George, Shefin S., Steele, Charles R., Ricci, Anthony J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8255937/
https://www.ncbi.nlm.nih.gov/pubmed/34258597
http://dx.doi.org/10.1016/j.xpro.2021.100637
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author George, Shefin S.
Steele, Charles R.
Ricci, Anthony J.
author_facet George, Shefin S.
Steele, Charles R.
Ricci, Anthony J.
author_sort George, Shefin S.
collection PubMed
description 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).
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spelling pubmed-82559372021-07-12 A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells George, Shefin S. Steele, Charles R. Ricci, Anthony J. STAR Protoc Protocol 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). Elsevier 2021-06-30 /pmc/articles/PMC8255937/ /pubmed/34258597 http://dx.doi.org/10.1016/j.xpro.2021.100637 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
George, Shefin S.
Steele, Charles R.
Ricci, Anthony J.
A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells
title A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells
title_full A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells
title_fullStr A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells
title_full_unstemmed A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells
title_short A two-photon FRAP protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells
title_sort two-photon frap protocol to measure the stereociliary membrane diffusivity in rat cochlear hair cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8255937/
https://www.ncbi.nlm.nih.gov/pubmed/34258597
http://dx.doi.org/10.1016/j.xpro.2021.100637
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