Flat-Field Super-Resolution Localization Microscopy with a Low-Cost Refractive Beam-Shaping Element.
View / Open Files
Authors
Ramirez, Pedro P Vallejo
Scherer, Katharina M
Publication Date
2018-04-04Journal Title
Sci Rep
ISSN
2045-2322
Publisher
Springer Science and Business Media LLC
Volume
8
Issue
1
Pages
5630
Language
eng
Type
Article
Physical Medium
Electronic
Metadata
Show full item recordCitation
Rowlands, C. J., Ströhl, F., Ramirez, P. P. V., Scherer, K. M., & Kaminski, C. F. (2018). Flat-Field Super-Resolution Localization Microscopy with a Low-Cost Refractive Beam-Shaping Element.. Sci Rep, 8 (1), 5630. https://doi.org/10.1038/s41598-018-24052-4
Abstract
Super-resolution single-molecule localization microscopy, often referred to as PALM/STORM, works by ensuring that fewer than one fluorophore in a diffraction-limited volume is emitting at any one time, allowing the observer to infer that the emitter is located at the center of the point-spread function. This requires careful control over the incident light intensity in order to control the rate at which fluorophores are switched on; if too many fluorophores are activated, their point-spread functions overlap, which impedes efficient localization. If too few are activated, the imaging time is impractically long. There is therefore considerable recent interest in constructing so-called 'top-hat' illumination profiles that provide a uniform illumination over the whole field of view. We present the use of a single commercially-available low-cost refractive beamshaping element that can be retrofitted to almost any existing microscope; the illumination profile created by this element demonstrates a marked improvement in the power efficiency of dSTORM microscopy, as well as a significant reduction in the propensity for reconstruction artifacts, compared to conventional Gaussian illumination.
Sponsorship
Wellcome Trust (089703/Z/09/Z)
Biotechnology and Biological Sciences Research Council (BB/H023917/1)
Engineering and Physical Sciences Research Council (EP/H018301/1)
Medical Research Council (MR/K02292X/1)
Medical Research Council (G0902243)
Medical Research Council (MR/K015850/1)
Wellcome Trust (093831/B/10/Z)
Wellcome Trust (203249/Z/16/Z)
Engineering and Physical Sciences Research Council (EP/L015889/1)
Identifiers
External DOI: https://doi.org/10.1038/s41598-018-24052-4
This record's URL: https://www.repository.cam.ac.uk/handle/1810/279536
Rights
Attribution 4.0 International (CC BY 4.0)
Licence URL: https://creativecommons.org/licenses/by/4.0/
Statistics
Total file downloads (since January 2020). For more information on metrics see the
IRUS guide.
Recommended or similar items
The current recommendation prototype on the Apollo Repository will be turned off on 03 February 2023. Although the pilot has been fruitful for both parties, the service provider IKVA is focusing on horizon scanning products and so the recommender service can no longer be supported. We recognise the importance of recommender services in supporting research discovery and are evaluating offerings from other service providers. If you would like to offer feedback on this decision please contact us on: support@repository.cam.ac.uk