Open AccessComparing strategies for deep astigmatism-based single-molecule localization microscopy
Author(s) -
Marijn Siemons,
Bas M. C. Cloin,
Desireé Salas,
Wilco Nijenhuis,
Eugene A. Katrukha,
Lukas C. Kapitein
Publication year - 2020
Publication title -
biomedical optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.382023
Subject(s) - optics , microscopy , astigmatism , adaptive optics , lens (geology) , numerical aperture , image resolution , physics , superresolution , materials science , computer science , artificial intelligence , wavelength , image (mathematics)
Single-molecule localization microscopy (SMLM) enables fluorescent microscopy with nanometric resolution. While localizing molecules close to the coverslip is relatively straightforward using high numerical aperture (NA) oil immersion (OI) objectives, optical aberrations impede SMLM deeper in watery samples. Adaptive optics (AO) with a deformable mirror (DM) can be used to correct such aberrations and to induce precise levels of astigmatism to encode the z- position of molecules. Alternatively, the use of water immersion (WI) objectives might be sufficient to limit the most dominant aberrations. Here we compare SMLM at various depths using either WI or OI with or without AO. In addition, we compare the performance of a cylindrical lens and a DM for astigmatism-based z- encoding. We find that OI combined with adaptive optics improves localization precision beyond the performance of WI-based imaging and enables deep (>10 µm) 3D localization.