Analysis of TrkB Receptor Activity Using FRET Sensors


Book Section

© 2018, Springer Science+Business Media New York. Here, we describe the use of 2-photon fluorescence lifetime imaging (2pFLIM) of a Förster resonance energy transfer (FRET) sensor to study the spatial and temporal activity pattern of the BDNF receptor TrkB. Combining 2pFLIM with laser uncaging of MNI-glutamate over single dendritic spines in organotypic hippocampal slices allows measurement of glutamate-induced changes in TrkB activity within a single spine or dendrite. This protocol covers the installation and setup of requisite hardware and software, the use of the software to acquire data, and the analysis of the data. A prerequisite for attempting this protocol is proficiency with 2-photon imaging of live cells and a microscope that is controlled by ScanImage 3.8 (Vidrio Technologies).

Full Text

Duke Authors

Cited Authors

  • Hall, CE; McNamara, JO; Yasuda, R

Published Date

  • January 1, 2019

Volume / Issue

  • 143 /

Book Title

  • Neuromethods

Start / End Page

  • 149 - 157

Digital Object Identifier (DOI)

  • 10.1007/7657_2018_12

Citation Source

  • Scopus