Method Article

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

May 29th, 2025

In This Article

Abstract

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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

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

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Results

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Rotary motion setup with foam ball, motor, rotary encoder; schematic and photo views.

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Two-photon microscopeN/AN/A
ScanImage 3.6Svoboda Lab/Vidrio TechnologyN/A
FLIMview MATLAB softwareN/AN/A
16x 0.8 NA water-immersion
objective		NikonMRP07220
AnimalTracker MATLAB softwareN/AN/A
Stereotaxic alignment systsemDavid kopf1900
Virus: tAKARα (AAV2/1 hSyntAKARα-WPRE)Addgene119921
HeadplateN/AN/A
Headplate holderN/AN/A
Circular coverslip (5 mm diameter)r) VWR101413-528

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Tags

Protein Kinase A ActivityTwo Photon FLIMFluorescence Lifetime ImagingHead Fixed MiceCranial Window ImagingEnforced LocomotionFRET Reporter AnalysisZ Stack AcquisitionLifetime Estimation ErrorRegion of Interest Tracking

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