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

• Main Authors: Allgeyer, E.S (Author), Cheng, J. (Author), Johnston, D.S (Author), Sirinakis, G. (Author)
• Format: Article
• Language: English
• Published: Optica Publishing Group (formerly OSA) 2022
• Subjects: Binding peptide; Biological imaging; Drosophilla; Geometry; Imaging speed; Optical resolving power; Paint; Proteins; Quantitative comparison; Spinning disks; Spinning-disk geometry; Super resolution imaging; Super-resolution microscopy; Thick samples
• Online Access: View Fulltext in Publisher

 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. © 2022 OSA - The Optical Society. All rights reserved.