Pupil function design for multifocal confocal, STED, and isoSTED microscopy
Open Access
- 16 June 2021
- journal article
- research article
- Published by Optica Publishing Group in Applied Optics
- Vol. 60 (18), 5354-5359
- https://doi.org/10.1364/ao.416585
Abstract
Point scanning super-resolution microscopy techniques such as stimulated emission depletion (STED) microscopy are powerful tools to observe biological samples at sub-diffraction limited resolution in three dimensions. However, scanning the sample with only a single beam limits the imaging speed in these microscopes. Here, we propose a concept to increase this speed by introducing highly flexible multifocal illumination and detection. We introduce phase patterns in the objectives' pupil planes to create arrays of foci in the sample plane with negligible loss of laser power. High uniformity of these foci's intensities is achieved by iteratively applying a weighted Gerchberg-Saxton phase retrieval algorithm. We characterize the performance of this iterative approach numerically and present simulation results that demonstrate the high quality of the focus arrays for future implementations in laser-scanning STED and isoSTED microscopes. The same approach can also be applied in diffraction-limited confocal laser scanning microscopy. Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License.Funding Information
- Wellcome Trust (203285/B/16/Z)
This publication has 33 references indexed in Scilit:
- Adaptive optics enables 3D STED microscopy in aberrating specimensOptics Express, 2012
- Optical Nanoscopy: From Acquisition to AnalysisAnnual Review of Biomedical Engineering, 2012
- Effects of polarization on the de-excitation dark focal spot in STED microscopyJournal of Optics, 2010
- STED microscopy reveals crystal colour centres with nanometric resolutionNature Photonics, 2009
- Microscopy and its focal switchNature Methods, 2008
- Comparison of I5M and 4Pi‐microscopyJournal of Microscopy, 2006
- Breaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteinsProceedings of the National Academy of Sciences of the United States of America, 2005
- Phase‐retrieved pupil functions in wide‐field fluorescence microscopyJournal of Microscopy, 2004
- Focal Spots of SizeOpen Up Far-Field Florescence Microscopy at 33 nm Axial ResolutionPhysical Review Letters, 2002
- Electromagnetic diffraction in optical systems, II. Structure of the image field in an aplanatic systemProceedings of the Royal Society of London. Series A - Mathematical and Physical Sciences, 1959