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Quantitative comparison of spinning disk geometries for PAINT based super-resolution microscopy.

Published version
Peer-reviewed

Type

Article

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Authors

Cheng, Jinmei 
St Johnston, Daniel 

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.

Description

Keywords

Journal Title

Biomed Opt Express

Conference Name

Journal ISSN

2156-7085
2156-7085

Volume Title

Publisher

Optica Publishing Group
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)