Constructing a cost-efficient, high-throughput and high-quality single-molecule localization microscope for super-resolution imaging.
Open accessAbstract
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.
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The technique used to quantify soluble Aβ aggregates has previously been validated in multiple studies and specifically detects aggregates, not monomers.
“The technique used to quantify soluble aggregates has previously been validated in multiple studies and has been shown to specifically detect aggregates, and not monomers [29–31].”
From Selective suppression of oligodendrocyte-derived amyloid beta rescues neuronal dysfunction in Alzheimer’s disease (2024) · cites this paper as [31]
Figure 5 (p. 56) describes imaging recombinant α-synuclein aggregates, while the Introduction (p. 1) mentions amyloid aggregates generally. Our copy does not address validation of a technique for quantifying soluble Aβ aggregates or show that it detects aggregates rather than monomers; searches for those terms found no such passage. (LLM, conf=0.76)
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Rajani, R. M., Ellingford, R., Hellmuth, M., Harris, S. S., Taso, O. S., Graykowski, D., Lam, F. K. W., Arber, C., Fertan, E., Danial, J. S. H., et al. (2024) · PLOS Biology · doi:10.1371/journal.pbio.3002727 · Cited by 57 · see citing works
“The technique used to quantify soluble aggregates has previously been validated in multiple studies and has been shown to specifically detect aggregates, and not monomers [29–31].”
BackgroundMethodSelf-citation
- Authors
- Danial, J. S. H., Lam, J. Y. L., Wu, Y., Woolley, M., Dimou, E., Cheetham, M. R., Emin, D., & Klenerman, D.
- Affiliations
- University of Cambridge (GB)
- Corresponding author
- Danial, John S. H.
- Klenerman, David
- ORCID
- Venue
- Nature Protocols
- Published
- 2022-08-24
- Volume
- 17
- Issue
- 11
- Pages
- 2570-2619
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W4293004995
- Cited by
- 40
- References
- 49
- Citations by year
- 2026: 3 · 2025: 11 · 2024: 12 · 2023: 14
- FWCI
- 4.08
- Type
- journal article
- Language
- en
- Update notices
- correction (2023-09-27)
- Open access
- Licence
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Apollo (University of Cambridge) (submittedVersion, cc-by)
- Apollo (cc-by)
- Funders
- University of Cambridge | King’s College Cambridge, University of Cambridge
- RCUK | Medical Research Council
- Royal Society
- Croucher Foundation
- Deutsche Forschungsgemeinschaft
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Force Microscopy Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
- Keywords
- single-molecule localization microscopy, super-resolution imaging, microscope construction, low-cost instrumentation, high-throughput imaging
- MeSH terms
- Single Molecule Imaging, Humans, Microscopy
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