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Updated: Jul 15, 2025
Simultaneous Two-photon In Vivo Imaging of Synaptic Inputs and Postsynaptic Targets in the Mouse Retrosplenial Cortex
Published on: March 13, 2016
Simultaneous multimodal three-photon and optical coherence microscopy of the mouse brain in the 1700 nm optical
Xusan Yang1,2, Siyang Liu3, Fei Xia1,4,5
1School of Applied and Engineering Physics, Cornell University, Ithaca, NY, 14853, USA.
Abstract:
Multimodal microscopy combining various imaging approaches can provide complementary information about tissue in a single imaging session. Here, we demonstrate a multimodal approach combining three-photon microscopy (3PM) and spectral-domain optical coherence microscopy (SD-OCM). We show that an optical parametric chirped-pulse amplification (OPCPA) laser source, which is the standard source for three-photon fluorescence excitation and third harmonic generation (THG), can be used for simultaneous OCM, 3-photon (3P) fluorescence and THG imaging. We validated the system performance in deep mouse brains in vivo with an OPCPA source operating at 1620 nm center wavelength. We visualized small structures such as myelinated axons, neurons, and large fiber tracts in white matter with high spatial resolution non-invasively using linear and nonlinear contrast at >1 mm depth in intact adult mouse brain. Our results showed that simultaneous OCM and 3PM at the long wavelength window can be conveniently combined for deep tissue imaging in vivo.
