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  -