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Constructing a cost-efficient, high-throughput and high-quality single-molecule localization microscope for super-resolution imaging.

Accepted version
Peer-reviewed

Type

Article

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Authors

Wu, Yunzhao 
Woolley, Matthew 

Abstract

Single-molecule localization microscopy (SMLM) leverages the power of modern optics to unleash ultra-precise structural nanoscopy of complex biological machines in their native environments as well as ultra-sensitive and high-throughput medical diagnostics with the sensitivity of a single molecule. To achieve this remarkable speed and resolution, SMLM setups are either built by research laboratories with strong expertise in optical engineering or commercially sold at a hefty price tag. The inaccessibility of SMLM to life scientists for technical or financial reasons is detrimental to the progress of biological and biomedical discoveries reliant on super-resolution imaging. In this work, we present the NanoPro, an economic, high-throughput, high-quality and easy-to-assemble SMLM for super-resolution imaging. We show that our instrument performs similarly to the most expensive, best-in-class commercial microscopes and rivals existing open-source microscopes at a lower price and construction complexity. To facilitate its wide adoption, we compiled a step-by-step protocol, accompanied by extensive illustrations, to aid inexperienced researchers in constructing the NanoPro as well as assessing its performance by imaging ground-truth samples as small as 20 nm. The detailed visual instructions make it possible for students with little expertise in microscopy engineering to construct, validate and use the NanoPro in <1 week, provided that all components are available.

Description

Keywords

Humans, Single Molecule Imaging, Microscopy

Journal Title

Nat Protoc

Conference Name

Journal ISSN

1754-2189
1750-2799

Volume Title

Publisher

Springer Science and Business Media LLC
Sponsorship
UK Dementia Research Institute (PILOT2020-DANIAL)
European Research Council (669237)
EPSRC (EP/T01427X/1)