We present a method for Z-super-localization in fluorescence microscopy, based on conical diffraction. By using a thin biaxial crystal, the Point Spread Function (PSF) shape of an objective is made to depend strongly on the z coordinate. This z dependence is then exploited to localize fluorescent emitters axially with a great precision. We study how this method can be used for single molecule imaging with a global assessment by Fisher information analysis. Preliminary experiments demonstrate that this technique can obtain resolutions of tens of nm with the use of high NA objectives.

Date Published: 9 March 2015
PDF: 11 pages
Proc. SPIE 9331, Single Molecule Spectroscopy and Superresolution Imaging VIII, 93310Q (9 March 2015); doi: 10.1117/12.2077712

Published in SPIE Proceedings Vol. 9331:
Single Molecule Spectroscopy and Superresolution Imaging VIII
Jörg Enderlein; Ingo Gregor; Zygmunt Karol Gryczynski; Rainer Erdmann; Felix Koberling, Editor(s)