Optimized localization analysis for single-molecule tracking and super-resolution microscopy.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 20364147.
- Also identified by DOI 10.1038/nmeth.1447 and PMC identifier 3127582.
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Abstract
We optimally localized isolated fluorescent beads and molecules imaged as diffraction-limited spots, determined the orientation of molecules and present reliable formulas for the precision of various localization methods. Both theory and experimental data showed that unweighted least-squares fitting of a Gaussian squanders one-third of the available information, a popular formula for its precision exaggerates beyond Fisher's information limit, and weighted least-squares may do worse, whereas maximum-likelihood fitting is practically optimal.
Medical subject headings
- Image Interpretation, Computer-Assisted
- Microscopy, Fluorescence