RT Journal Article
SR Electronic
T1 Direct Supercritical Angle Localization Microscopy for Nanometer 3D Superresolution
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.25.171058
DO 10.1101/2020.06.25.171058
A1 Anindita Dasgupta
A1 Joran Deschamps
A1 Ulf Matti
A1 Uwe Hübner
A1 Jan Becker
A1 Sebastian Strauss
A1 Ralf Jungmann
A1 Rainer Heintzmann
A1 Jonas Ries
YR 2020
UL http://biorxiv.org/content/early/2020/06/26/2020.06.25.171058.abstract
AB 3D single molecule localization microscopy (SMLM) is an emerging superresolution method for structural cell biology, as it allows probing precise positions of proteins in cellular structures. Supercritical angle fluorescence strongly depends on the z-position of the fluorophore and can be used for z localization in a method called supercritical angle localization microscopy (SALM). Here, we realize the full potential of SALM by directly splitting supercritical and undercritical emission, using an ultra-high NA objective, and applying new fitting routines to extract precise intensities of single emitters, resulting in a four-fold improved z-resolution compared to the state of the art. We demonstrate nanometer isotropic localization precision on DNA origami structures, and on clathrin coated vesicles and microtubules in cells, illustrating the potential of SALM for cell biology.Competing Interest StatementThe authors have declared no competing interest.