A hydrophilic gel matrix for single-molecule super-resolution microscopy
- 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.
Author: | Patrick J. M. Zessin, Carmen L. Krüger, Sebastian MalkuschORCiDGND, Ulrike EndesfelderORCiDGND, Mike HeilemannORCiDGND |
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URN: | urn:nbn:de:hebis:30:3-316232 |
DOI: | https://doi.org/10.1186/2192-2853-2-4 |
ISSN: | 2192-2853 |
Parent Title (English): | Optical Nanoscopy |
Publisher: | Springer Open |
Place of publication: | Berlin ; Heidelberg [u. a.] |
Document Type: | Article |
Language: | English |
Year of Completion: | 2013 |
Date of first Publication: | 2013/09/10 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2013/09/18 |
Tag: | Drift correction; Registration; Single-molecule localization microscopy; Super-resolution microscopy |
Volume: | 2 |
Issue: | 4 |
Page Number: | 8 |
First Page: | 1 |
Last Page: | 8 |
Note: | © 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. |
HeBIS-PPN: | 353192600 |
Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
Sammlungen: | Universitätspublikationen |
Licence (German): | Creative Commons - Namensnennung 2.0 |