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Regensburg 2022 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 9: Bioimaging

BP 9.6: Vortrag

Dienstag, 6. September 2022, 11:00–11:15, H16

An open-top scanned oblique lightsheet microscope for neuronal network imaging — •Achim Theo Brinkop1, Stefan Stöberl1, Florian Schorre1, and Friedhelm Serwane1,2,31Faculty of Physics, LMU Munich, Germany — 2Munich Cluster for Systems Neurology (SyNergy), Germany — 3Graduate School of Systemic Neuroscience (GSN), Munich, Germany

Understanding signal processing in neuronal networks such as brain organoids on a single-neuron level has remained a challenge. Imaging network activity requires a millisecond temporal resolution with single-neuron spatial resolution, all in an observation volume containing the 3D network. Advances in lightsheet microscopy have brought this goal closer to experimental reach, but at the cost of complex optical set-ups which (i) impose geometrical constraints to sample mounting or (ii) require multiple imaging objectives with custom optical components.

We report on the development of an open-top single-objective oblique lightsheet microscope which reduces the complexity compared to existing set-ups. We implement the open-top geometry by using only two primary objectives. The lightsheet is digital scanned by a fast galvo mirror to maintain high image quality. Our first prototype with excitation wavelengths of 561 (488, 638) nm is expected to allow for a 1/e2-resolution of 1.84 (1.47, 1.95) µm axially and 0.27 (0.23, 0.30) µm laterally. It offers a volumetric temporal resolution of 8 (2) Hz for a volume of 400 x 90 (360) x 100 µm3.

With this set-up, we aim to gain insights into large neuronal networks of retina organoids, both in wildtype and disease condition.

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