Imaging through the Whole Brain of Drosophila at λ/20 Super-resolution
Han-Yuan Lin,
Li-An Chu,
Hsuan Yang,
Kuo-Jen Hsu,
Yen-Yin Lin,
Keng-Hui Lin,
Shi-Wei Chu,
Ann-Shyn Chiang
Affiliations
Han-Yuan Lin
Department of Physics, National Taiwan University, Taipei 10617, Taiwan
Li-An Chu
Brain Research Center, National Tsing Hua University, Hsinchu 30013, Taiwan; Institute of Physics, Academia Sinica, Taipei 11529, Taiwan
Hsuan Yang
Institute of Physics, Academia Sinica, Taipei 11529, Taiwan
Kuo-Jen Hsu
Department of Physics, National Taiwan University, Taipei 10617, Taiwan; Brain Research Center, National Tsing Hua University, Hsinchu 30013, Taiwan
Yen-Yin Lin
Brain Research Center, National Tsing Hua University, Hsinchu 30013, Taiwan
Keng-Hui Lin
Institute of Physics, Academia Sinica, Taipei 11529, Taiwan; Corresponding author
Shi-Wei Chu
Department of Physics, National Taiwan University, Taipei 10617, Taiwan; Molecular Imaging Center, National Taiwan University, Taipei 10617, Taiwan; Corresponding author
Ann-Shyn Chiang
Brain Research Center, National Tsing Hua University, Hsinchu 30013, Taiwan; Institute of Physics, Academia Sinica, Taipei 11529, Taiwan; Institute of Systems Neuroscience, National Tsing Hua University, Hsinchu 30013, Taiwan; Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli 35053, Taiwan; Kavli Institute for Brain and Mind, University of California, San Diego, La Jolla, CA 92093-0526, USA; Corresponding author
Summary: Recently, many super-resolution technologies have been demonstrated, significantly affecting biological studies by observation of cellular structures down to nanometer precision. However, current super-resolution techniques mostly rely on wavefront engineering or wide-field imaging of signal blinking or fluctuation, and thus imaging depths are limited due to tissue scattering or aberration. Here we present a technique that is capable of imaging through an intact Drosophila brain with 20-nm lateral resolution at ∼200 μm depth. The spatial resolution is provided by molecular localization of a photoconvertible fluorescent protein Kaede, whose red form is found to exhibit blinking state. The deep-tissue observation is enabled by optical sectioning of spinning disk microscopy, as well as reduced scattering from optical clearing. Together these techniques are readily available for many biologists, providing three-dimensional resolution of densely entangled dendritic fibers in a complete Drosophila brain. The method paves the way toward whole-brain neural network studies and is applicable to other high-resolution bioimaging. : Optical Imaging; Neuroscience; Techniques in Neuroscience Subject Areas: Optical Imaging, Neuroscience, Techniques in Neuroscience
