Format

Send to

Choose Destination
Sci Rep. 2014 Jan 24;4:3854. doi: 10.1038/srep03854.

The fidelity of stochastic single-molecule super-resolution reconstructions critically depends upon robust background estimation.

Author information

1
1] Section of Molecular Cytology and Van Leeuwenhoek Centre of Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam Science Park 904, NL-1098 XH Amsterdam The Netherlands [2].
2
1] Section of Molecular Cytology and Van Leeuwenhoek Centre of Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam Science Park 904, NL-1098 XH Amsterdam The Netherlands [2] [3].
3
1] Division of Cell Biology and Van Leeuwenhoek Centre of Advanced Microscopy, The Netherlands Cancer Institute, Amsterdam, The Netherlands [2].
4
Section of Molecular Cytology and Van Leeuwenhoek Centre of Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam Science Park 904, NL-1098 XH Amsterdam The Netherlands.
5
Division of Cell Biology and Van Leeuwenhoek Centre of Advanced Microscopy, The Netherlands Cancer Institute, Amsterdam, The Netherlands.

Abstract

The quality of super resolution images obtained by stochastic single-molecule microscopy critically depends on image analysis algorithms. We find that the choice of background estimator is often the most important determinant of reconstruction quality. A variety of techniques have found use, but many have a very narrow range of applicability depending upon the characteristics of the raw data. Importantly, we observe that when using otherwise accurate algorithms, unaccounted background components can give rise to biases on scales defeating the purpose of super-resolution microscopy. We find that a temporal median filter in particular provides a simple yet effective solution to the problem of background estimation, which we demonstrate over a range of imaging modalities and different reconstruction methods.

Comment in

PMID:
24458236
PMCID:
PMC3900998
DOI:
10.1038/srep03854
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center