Multicolor fiber-optic two-photon endomicroscopy for brain imaging

Visualizing activity patterns of distinct cell types during complex behaviors is essential to understand complex neural networks. It remains challenging to excite multiple fluorophores simultaneously so that different types of neurons can be imaged. In this Letter, we report a multicolor fiber-optic two-photon endomicroscopy platform in which two pulses from a Ti:sapphire laser and an optical parametric oscillator were synchronized and delivered through a single customized double-clad fiber to excite multiple chromophores. A third virtual wavelength could also be generated by spatial-temporal overlapping of the two pulses. The performance of the fiber-optic multicolor two-photon endomicroscope was demonstrated by in vivo imaging of a mouse cerebral cortex with “Brainbow” labeling.