We report a two-photon excitation fluorescence microscopy imager that uses a fiber-delivered fs supercontinuum (SC) generation source to achieve simultaneous dual-wavelength imaging. The SC generation is based on pumping a polarization-maintaining photonic crystal fiber with a fiber laser. The SC is then delivered to the imaging head through a broadband hollow-core fiber. Excitation wavelengths centered at 920 nm and 1180 nm were selected, which enabled simultaneous imaging of neurons transfected with GCaMP6 and administrated with muscimol-conjugated BODIPY. Imaging results of calcium dynamics of in vivo mouse brain and cultured primary neurons were collected to demonstrate the technology feasibility.
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