Quantitative comparison of spinning disk geometries for PAINT based super-resolution microscopy.
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Publication Date
2022-07-01Journal Title
Biomed Opt Express
ISSN
2156-7085
Publisher
Optica Publishing Group
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Sirinakis, G., Allgeyer, E. S., Cheng, J., & St Johnston, D. (2022). Quantitative comparison of spinning disk geometries for PAINT based super-resolution microscopy.. Biomed Opt Express https://doi.org/10.1364/BOE.459490
Abstract
PAINT methods that use DNA- or protein- based exchangeable probes have become popular for super-resolution imaging and have been combined with spinning disk confocal microscopy for imaging thicker samples. However, the widely available spinning disks used for routine biological imaging are not optimized for PAINT-based applications and may compromise resolution and imaging speed. Here, we use Drosophila egg chambers in the presence of the actin-binding peptide Lifeact to study the performance of four different spinning disk geometries. We find that disk geometries with higher light collection efficiency perform better for PAINT-based super-resolution imaging due to increased photon numbers and, subsequently, detection of more blinking events.
Sponsorship
Wellcome Trust (095927/B/11/Z)
Biotechnology and Biological Sciences Research Council (BB/P026486/1)
Wellcome Trust (203285/Z/16/Z)
Wellcome Trust (092096/Z/10/Z)
Wellcome Trust (203144/Z/16/Z)
Wellcome Trust (080007/Z/06/Z)
Wellcome Trust (080007/B/06/Z)
Wellcome Trust (207496/Z/17/Z)
Wellcome Trust (207496/Z/17/A)
Cancer Research UK (C6946/A24843)
Cancer Research Uk (None)
Identifiers
External DOI: https://doi.org/10.1364/BOE.459490
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336955
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