Journal
BIOMEDICAL OPTICS EXPRESS
Volume 13, Issue 7, Pages 3773-3785Publisher
Optica Publishing Group
DOI: 10.1364/BOE.459490
Keywords
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Funding
- Wellcome Trust [080007, 092096, 203144, 203285, 207496]
- Biotechnology and Biological Sciences Research Council [BB/P026486/1]
- Cancer Research UK [A14492, A24823, C6946/A24843]
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When using spinning disks combined with PAINT methods for super-resolution imaging, the geometry of the spinning disk has an impact on imaging performance. Disk geometries with higher light collection efficiency perform better for PAINT-based super-resolution imaging.
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.
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