Open AccessOptical properties of adult Drosophila brains in one-, two-, and three-photon microscopy
Author(s) -
Kuan-Lun Hsu,
Yen-Yin Lin,
AnnShyn Chiang,
ShiWei Chu
Publication year - 2019
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.10.001627
Subject(s) - optics , scattering , light scattering , two photon excitation microscopy , wavelength , connectome , distortion (music) , photon , excitation , microscopy , physics , materials science , optoelectronics , biology , neuroscience , fluorescence , amplifier , cmos , quantum mechanics , functional connectivity
Drosophila is widely used in connectome studies due to its small brain size, sophisticated genetic tools, and the most complete single-neuron-based anatomical brain map. Surprisingly, even the brain thickness is only 200-μm, common Ti:sapphire-based two-photon excitation cannot penetrate, possibly due to light aberration/scattering of trachea. Here we quantitatively characterized scattering and light distortion of trachea-filled tissues, and found that trachea-induced light distortion dominates at long wavelength by comparing one-photon (488-nm), two-photon (920-nm), and three-photon (1300-nm) excitations. Whole- Drosophila -brain imaging is achieved by reducing tracheal light aberration/scattering via brain-degassing or long-wavelength excitation at 1300-nm. Our work paves the way toward constructing whole-brain connectome in a living Drosophila .