PT  - JOURNAL ARTICLE
AU  - Dasgupta, Anindita
AU  - Deschamps, Joran
AU  - Matti, Ulf
AU  - Hübner, Uwe
AU  - Becker, Jan
AU  - Strauss, Sebastian
AU  - Jungmann, Ralf
AU  - Heintzmann, Rainer
AU  - Ries, Jonas
TI  - Direct Supercritical Angle Localization Microscopy for Nanometer 3D Superresolution
AID  - 10.1101/2020.06.25.171058
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.06.25.171058
4099  - http://biorxiv.org/content/early/2020/06/26/2020.06.25.171058.short
4100  - http://biorxiv.org/content/early/2020/06/26/2020.06.25.171058.full
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.