TY - JOUR A1 - Zessin, Patrick J. M. A1 - Krüger, Carmen L. A1 - Malkusch, Sebastian A1 - Endesfelder, Ulrike A1 - Heilemann, Mike T1 - A hydrophilic gel matrix for single-molecule super-resolution microscopy T2 - Optical Nanoscopy N2 - Background: Novel microscopic techniques which bypass the resolution limit in light microscopy are becoming routinely established today. The higher spatial resolution of super-resolution microscopy techniques demands for precise correction of drift, spectral and spatial offset of images recorded at different axial planes. Methods: We employ a hydrophilic gel matrix for super-resolution microscopy of cellular structures. The matrix allows distributing fiducial markers in 3D, and using these for drift correction and multi-channel registration. We demonstrate single-molecule super-resolution microscopy with photoswitchable fluorophores at different axial planes. We calculate a correction matrix for each spectral channel, correct for drift, spectral and spatial offset in 3D. Results and discussion: We demonstrate single-molecule super-resolution microscopy with photoswitchable fluorophores in a hydrophilic gel matrix. We distribute multi-color fiducial markers in the gel matrix and correct for drift and register multiple imaging channels. We perform two-color super-resolution imaging of click-labeled DNA and histone H2B in different axial planes, and demonstrate the quality of drift correction and channel registration quantitatively. This approach delivers robust microscopic data which is a prerequisite for data interpretation. KW - Single-molecule localization microscopy KW - Super-resolution microscopy KW - Drift correction KW - Registration Y1 - 2013 UR - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/31623 UR - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-316232 SN - 2192-2853 N1 - © 2013 Zessin et al.; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. VL - 2 IS - 4 SP - 1 EP - 8 PB - Springer Open CY - Berlin ; Heidelberg [u. a.] ER -