Varying label density allows artifact-free analysis of membrane-protein nanoclusters.
basic_science · Level V
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- Record sourced from PubMed, PMID 27295310.
- Also identified by DOI 10.1038/nmeth.3897 and PMC identifier 6404959.
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Abstract
We present a method to robustly discriminate clustered from randomly distributed molecules detected with techniques based on single-molecule localization microscopy, such as PALM and STORM. The approach is based on deliberate variation of labeling density, such as titration of fluorescent antibody, combined with quantitative cluster analysis, and it thereby circumvents the problem of cluster artifacts generated by overcounting of blinking fluorophores. The method was used to analyze nanocluster formation in resting and activated immune cells.
Medical subject headings
- Artifacts
- Cell Membrane
- Fluorescent Dyes
- Membrane Proteins
- Microscopy, Fluorescence
- Nanostructures