Open AccessPredicting resolution and image quality in RESOLFT and other point scanning microscopes [Invited]
Author(s) -
Andreas Bodén,
Xavier Casas Moreno,
Benjamin Cooper,
Andrew G. York,
Ilaria Testa
Publication year - 2020
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.389911
Subject(s) - photobleaching , point spread function , microscope , optics , fluorophore , optical transfer function , image quality , optical sectioning , resolution (logic) , microscopy , computer science , image resolution , point (geometry) , sampling (signal processing) , computer vision , physics , image (mathematics) , artificial intelligence , fluorescence , mathematics , geometry , detector
The performance of fluorescence microscopy and nanoscopy is often discussed by the effective point spread function and the optical transfer function. However, due to the complexity of the fluorophore properties such as photobleaching or other forms of photoswitching, which introduce a variance in photon emission, it is not trivial to choose optimal imaging parameters and to predict the spatial resolution. In this paper, we analytically derive a theoretical framework for estimating the achievable resolution of a microscope depending on parameters such as photoswitching, labeling densities, exposure time and sampling. We developed a numerical simulation software to analyze the impact of reversibly switchable probes in RESOLFT imaging.