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; Bi Chang Chen

doi:10.1038/s41467-019-12715-3

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^*, Bi Chang Chen

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

25 Citations (Scopus)

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.

Original language	English
Article number	4762
Journal	Nature Communications
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41467-019-12715-3
Publication status	Published - 2019 Dec 1
Externally published	Yes

ASJC Scopus subject areas

General Chemistry
General Biochemistry,Genetics and Molecular Biology
General Physics and Astronomy

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10.1038/s41467-019-12715-3

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title = "Rapid single-wavelength lightsheet localization microscopy for clarified tissue",

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.",

author = "Chu, {Li An} and Lu, {Chieh Han} and Yang, {Shun Min} and Liu, {Yen Ting} and Feng, {Kuan Lin} and Tsai, {Yun Chi} and Chang, {Wei Kun} and Wang, {Wen Cheng} and Chang, {Shu Wei} and Peilin Chen and Lee, {Ting Kuo} and Hwu, {Yeu Kuang} and Chiang, {Ann Shyn} and Chen, {Bi Chang}",

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doi = "10.1038/s41467-019-12715-3",

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AU - Chu, Li An

AU - Lu, Chieh Han

AU - Yang, Shun Min

AU - Liu, Yen Ting

AU - Feng, Kuan Lin

AU - Tsai, Yun Chi

AU - Chang, Wei Kun

AU - Wang, Wen Cheng

AU - Chang, Shu Wei

AU - Chen, Peilin

AU - Lee, Ting Kuo

AU - Hwu, Yeu Kuang

AU - Chiang, Ann Shyn

AU - Chen, Bi Chang

PY - 2019/12/1

Y1 - 2019/12/1

N2 - 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.

AB - 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.

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Rapid single-wavelength lightsheet localization microscopy for clarified tissue

Abstract

ASJC Scopus subject areas

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