Rapid reconstruction of neural circuits using tissue expansion and lattice light sheet microscopy

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eLife. 2022 Oct 26;. doi: 10.7554/eLife.81248

Rapid reconstruction of neural circuits using tissue expansion and lattice light sheet microscopy Saalfeld LabTillberg LabFlyLight

Lillvis Joshua, Otsuna Hideo, Ding Xiaoyu, Pisarev Igor, Kawase Takashi, Colonell Jennifer, Rokicki Konrad, Goina Cristian, Gao Ruixuan, Hu Amy, Wang Kaiyu, Bogovic John, Milkie Daniel, Boyden Edward, Saalfeld Stephan, Tillberg Paul, Dickson Barry

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Abstract

Electron microscopy (EM) allows for the reconstruction of dense neuronal connectomes but suffers from low throughput, limiting its application to small numbers of reference specimens. We developed a protocol and analysis pipeline using tissue expansion and lattice light-sheet microscopy (ExLLSM) to rapidly reconstruct selected circuits across many samples with single synapse resolution and molecular contrast. We validate this approach in Drosophila, demonstrating that it yields synaptic counts similar to those obtained by EM, can be used to compare counts across sex and experience, and to correlate structural connectivity with functional connectivity. This approach fills a critical methodological gap in studying variability in the structure and function of neural circuits across individuals within and between species.