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Two-Photon FLIM with a Femtosecond Fibre Laser

We demonstrate two-photon FLIM with a femtosecond fibre laser. Our system consists of a bh DCS-120 scanner attached to a Nikon TE microscope, a 785 nm Toptica Femto Fibre Pro laser, HPM-100 hybrid detectors in a non-descanned beam configuration, and a TCSPC FLIM system with two SPC-150NX TCSPC modules, a GVD-120 scan controller, and a DCC-100 detector controller. We show that the system is perfectly suited to record NADH FLIM data from cells and tissue. It also records autofluorescence images from small organisms. FLIM images are also obtained from stained specimens, such as the Invitrogen BPAE cell and mouse kidney samples.

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