Open AccessGCaMP6 ΔF/F dependence on the excitation wavelength in 3-photon and 2-photon microscopy of mouse brain activity
Author(s) -
Dimitre G. Ouzounov,
Tianyu Wang,
Chunyan Wu,
Chris Xu
Publication year - 2019
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.10.003343
Subject(s) - two photon excitation microscopy , excitation , fluorescence , photon , optics , fluorescence lifetime imaging microscopy , wavelength , physics , microscopy , photon counting , in vivo , calcium imaging , laser , excitation wavelength , materials science , nuclear magnetic resonance , preclinical imaging , calcium , biology , microbiology and biotechnology , quantum mechanics , metallurgy
A fundamental challenge in calcium imaging is to minimize the excitation laser power while still maintaining a sufficient signal-to-noise ratio to distinguish individual transients in the fluorescence traces. It is important to characterize relative fluorescence (i.e., ΔF/F) dependence on the excitation wavelength in vivo where the environment cannot be controlled effectively during imaging. Leveraging time division multiplexing of two excitation beams to achieve nearly simultaneous 2-photon and 3-photon imaging of the calcium transients, we measured systematically the ΔF/F dependence on the excitation wavelength in 2-photon and 3-photon in vivo imaging of neuronal activity in mouse brain labeled with GCaMP6s. The technique can be applied to in vivo measurements of other fluorescence sensors.