期刊
JOURNAL OF PHYSICS D-APPLIED PHYSICS
卷 52, 期 20, 页码 -出版社
IOP PUBLISHING LTD
DOI: 10.1088/1361-6463/ab0862
关键词
single-molecule localization microscopy; DNA-PAINT; super-resolution microscopy; fiducial marker based drift correction; single-particle tracking PALM; fluorescent particles
资金
- Max Planck Society
- SYNMIKRO
- Fonds der Chemischen Industrie
- IMPRS-Mic
- European Union [750673]
- Marie Curie Actions (MSCA) [750673] Funding Source: Marie Curie Actions (MSCA)
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.
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