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.
Original language | English |
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Journal | Nature Methods |
Volume | 7 |
Issue number | 5 |
Pages (from-to) | 377-U59 |
ISSN | 1548-7091 |
DOIs | |
Publication status | Published - 2010 |