Visualizing Protein Kinase A Activity in a Mouse Using Two-Photon Fluorescence Lifetime Imaging Microscopy

Abstract

Source: Jongbloets, B. C., et al. Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy. J. Vis. Exp. (2019)

This video demonstrates a two-photon fluorescence lifetime imaging microscopy procedure for visualizing protein kinase A activity in head-fixed, behaving mice during enforced locomotion.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

Results

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Two-photon microscope	N/A	N/A
ScanImage 3.6	Svoboda Lab/Vidrio Technology	N/A
FLIMview MATLAB software	N/A	N/A
16x 0.8 NA water-immersion objective	Nikon	MRP07220
AnimalTracker MATLAB software	N/A	N/A
Stereotaxic alignment systsem	David kopf	1900
Virus: tAKARα (AAV2/1 hSyntAKARα-WPRE)	Addgene	119921
Headplate	N/A	N/A
Headplate holder	N/A	N/A
Circular coverslip (5 mm diameter)	r) VWR	101413-528