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 Dasgupta, Anindita
A1 Deschamps, Joran
A1 Matti, Ulf
A1 Hübner, Uwe
A1 Becker, Jan
A1 Strauss, Sebastian
A1 Jungmann, Ralf
A1 Heintzmann, Rainer
A1 Ries, Jonas
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.