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Rapid single-wavelength lightsheet localization microscopy for clarified tissue

Author

Listed:
  • Li-An Chu

    (Brain Research Center, National Tsing Hua University
    Institute of Physics, Academia Sinica)

  • Chieh-Han Lu

    (Institute of Physics, Academia Sinica
    Research Center for Applied Sciences, Academia Sinica
    Department of Genetics and Complex Diseases, Harvard T H Chan School of Public Health)

  • 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
    Institute of Physics, Academia Sinica
    National Tsing Hua University
    Kaohsiung Medical University)

  • Bi-Chang Chen

    (Brain Research Center, National Tsing Hua University
    Research Center for Applied Sciences, Academia Sinica)

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.

Suggested Citation

  • 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 &, 2019. "Rapid single-wavelength lightsheet localization microscopy for clarified tissue," Nature Communications, Nature, vol. 10(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-12715-3
    DOI: 10.1038/s41467-019-12715-3
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    Cited by:

    1. Xuejiao Tian & Tzu-Yang Lin & Po-Ting Lin & Min-Ju Tsai & Hsin Chen & Wen-Jie Chen & Chia-Ming Lee & Chiao-Hui Tu & Jui-Cheng Hsu & Tung-Han Hsieh & Yi-Chung Tung & Chien-Kai Wang & Suewei Lin & Li-An, 2024. "Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy," Nature Communications, Nature, vol. 15(1), pages 1-16, December.

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