Rapid single-wavelength lightsheet localization microscopy for clarified tissue

Li-An Chu, Chieh-Han Lu, Shun-Min Yang, Yen-Ting Liu, Kuan-Lin Feng, Yun-Chi Tsai, Wei-Kun Chang, Wen-Cheng Wang, Shu-Wei Chang, Peilin Chen, Ting-Kuo Lee, Yeu-Kuang Hwu, Ann-Shyn Chiang () and Bi-Chang Chen ()
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Li-An Chu: Brain Research Center, National Tsing Hua University
Chieh-Han Lu: Institute of Physics, Academia Sinica
Shun-Min Yang: Institute of Physics, Academia Sinica
Yen-Ting Liu: Research Center for Applied Sciences, Academia Sinica
Kuan-Lin Feng: National Tsing Hua University
Yun-Chi Tsai: Research Center for Applied Sciences, Academia Sinica
Wei-Kun Chang: Brain Research Center, National Tsing Hua University
Wen-Cheng Wang: Research Center for Applied Sciences, Academia Sinica
Shu-Wei Chang: Research Center for Applied Sciences, Academia Sinica
Peilin Chen: Research Center for Applied Sciences, Academia Sinica
Ting-Kuo Lee: Institute of Physics, Academia Sinica
Yeu-Kuang Hwu: Institute of Physics, Academia Sinica
Ann-Shyn Chiang: Brain Research Center, National Tsing Hua University
Bi-Chang Chen: Brain Research Center, National Tsing Hua University

Nature Communications, 2019, vol. 10, issue 1, 1-10

Abstract: Abstract Optical super-resolution microscopy allows nanoscale imaging of protein molecules in intact biological tissues. However, it is still challenging to perform large volume super-resolution imaging for entire animal organs. Here we develop a single-wavelength Bessel lightsheet method, optimized for refractive-index matching with clarified specimens to overcome the aberrations encountered in imaging thick tissues. Using spontaneous blinking fluorophores to label proteins of interest, we resolve the morphology of most, if not all, dopaminergic neurons in the whole adult brain (3.64 × 107 µm3) of Drosophila melanogaster at the nanometer scale with high imaging speed (436 µm3 per second) for localization. Quantitative single-molecule localization reveals the subcellular distribution of a monoamine transporter protein in the axons of a single, identified serotonergic Dorsal Paired Medial (DPM) neuron. Large datasets are obtained from imaging one brain per day to provide a robust statistical analysis of these imaging data.

Date: 2019
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DOI: 10.1038/s41467-019-12715-3

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