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.

journal_name

Nat Commun

journal_title

Nature communications

authors

Chu LA,Lu CH,Yang SM,Liu YT,Feng KL,Tsai YC,Chang WK,Wang WC,Chang SW,Chen P,Lee TK,Hwu YK,Chiang AS,Chen BC

doi

10.1038/s41467-019-12715-3

subject

Has Abstract

pub_date

2019-10-18 00:00:00

pages

4762

issue

1

issn

2041-1723

pii

10.1038/s41467-019-12715-3

journal_volume

10

pub_type

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