Whole-Brain Super-Resolution Imaging in Drosophila

Shi-Wei Chu^1*

¹physics, National Taiwan University, Taipei, Taiwan

* Presenter:Shi-Wei Chu, email:swchu@phys.ntu.edu.tw

Due to strong scattering/aberration from neuron/trachea in living Drosophila brains, the penetration depth of two-photon-microscopy is surprisingly limited to ~100-μm. Here we demonstrated that for in vitro observation, degassing allows penetration through a whole brain, but for in vivo functional imaging, 1300-nm-excited three-photon-microscopy is required to provide whole-living-brain observation with better sectioning and less background. In addition, by combining optical clearing to reduce scattering, confocal microscopy to provide optical sectioning, a novel blinking photoconvertible fluorescent protein Kaede, and molecular localization techniques, 20 nm lateral resolution across the whole Drosophila brain is achieved. Our results pave the way toward the construction of whole-brain functional connectome with optical imaging modalities.

Keywords: brain , three-photon, localization, clearing