Balinovic, A;
Albrecht, D;
Endesfelder, U;
(2019)
Spectrally red-shifted fluorescent fiducial markers for optimal drift correction in localization microscopy.
Journal of Physics D - Applied Physics
, 52
, Article 204002. 10.1088/1361-6463/ab0862.
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Abstract
Precise drift correction is crucial for every single-molecule localization-based microscopy experiment. We evaluate commonly used fiducial markers such as gold nanoparticles, TetraSpeck microspheres, FluoSpheres and nanodiamonds. We introduce spectrally red-shifted fluorescent particles as optimal fiducial markers. Our concept exploits exciting the fluorescent particles at low efficiency far away from their absorption maximum. A fluorescent particle that is covered by a multitude of dyes will nevertheless yield a bright fiducial signal. This represents a simple yet powerful approach to alleviate common problems in drift corrections caused by photobleaching, sudden signal intensity changes or saturation effects of fiducial markers. We adapt our approach for PALM, sptPALM and DNA-PAINT experiments and demonstrate its simple use for varying imaging conditions in different spectral channels.
| Type: | Article |
|---|---|
| Title: | Spectrally red-shifted fluorescent fiducial markers for optimal drift correction in localization microscopy |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1088/1361-6463/ab0862 |
| Publisher version: | https://doi.org/10.1088/1361-6463/ab0862 |
| Language: | English |
| Additional information: | Original content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI. For more information see: http://creativecommons.org/licenses/by/3.0/ |
| Keywords: | Science & Technology, Physical Sciences, Physics, Applied, Physics, single-molecule localization microscopy, DNA-PAINT, super-resolution microscopy, fiducial marker based drift correction, single-particle tracking PALM, fluorescent particles, SINGLE-MOLECULE LOCALIZATION, LIVE ESCHERICHIA-COLI, RNA-POLYMERASE, SPATIAL-ORGANIZATION, SUPERRESOLUTION, CELL, REVEALS |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10072401 |
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