Berlin 2024 – scientific programme
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BP: Fachverband Biologische Physik
BP 15: Poster IIb
BP 15.18: Poster
Tuesday, March 19, 2024, 18:00–20:30, Poster F
Electro-optic imaging for a lightsheet based fluorescence lifetime imaging microscope (FLIM) — •Nils Bode1, Adam Bowman2, Dara Dowlatshahi3, Rose Knight3, Soichi Wakatsuki3, and Mark Kasevich2 — 1Physics Department, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstraße 1, 91058 Erlangen, Germany — 2Physics Department, Stanford University, 382 Via Pueblo Mall, Stanford, CA 94305, USA — 3School of Medicine, Stanford University, 318 Campus Drive West, Clark Center, Stanford, CA 94305, USA
Electro-optic imaging enables fast fluorescence lifetime microscopy with throughputs that are 103−105 times higher than those of typically used single photon counters. This enables the investigation of biological processes, like spatially resolved neuron activity and action potential propagation, that are beyond earlier systems capability. The presented electro-optic imaging setup utilizes a Pockels cell in combination with polarization splitting optics to acquire two distinct temporal gates per frame, whose ratio allows for lifetime calculation. For this work an electro-optical setup was built around a classical lightsheet microscope. This combination enables fast fluorescent lifetime imaging of volumetric samples. We used Arabidopsis thaliana seedlings, both labeled and label-free, as biological sample systems and performed spatial and lifetime calibration with fluorescent beads.
Keywords: lightsheet microscopy; fluorescence lifetime microscopy; electro optic imaging; polarization gating