Varying label density allows artifact-free analysis of membrane-protein nanoclusters

We present a new method to robustly discriminate clustered from random distributions of molecules detected with single molecule localization microscopy-based techniques like PALM and STORM. The approach is based on the deliberate variation of the labeling density, e.g. by titration of fluorescent an...

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Detalles Bibliográficos
Autores principales: Baumgart, Florian, Arnold, Andreas, Leskovar, Konrad, Staszek, Kaj, Fölser, Martin, Weghuber, Julian, Stockinger, Hannes, Schütz, Gerhard. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2016
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6404959/
https://www.ncbi.nlm.nih.gov/pubmed/27295310
http://dx.doi.org/10.1038/nmeth.3897
Descripción
Sumario:We present a new method to robustly discriminate clustered from random distributions of molecules detected with single molecule localization microscopy-based techniques like PALM and STORM. The approach is based on the deliberate variation of the labeling density, e.g. by titration of fluorescent antibody, combined with quantitative cluster analysis. It thereby circumvents the problem of cluster artifacts generated by overcounting of blinking fluorophores.

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