期刊
BIOPHYSICAL JOURNAL
卷 86, 期 2, 页码 1185-1200出版社
BIOPHYSICAL SOCIETY
DOI: 10.1016/S0006-3495(04)74193-4
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资金
- NIAID NIH HHS [R01 AI050747, R01 AI039167, R21 AI053748, R01 AI50747, R21 AI53748, R01 AI39167] Funding Source: Medline
One of the most basic questions in single-molecule microscopy concerns the accuracy with which the location of a single molecule can be determined. Using the Fisher information matrix it is shown that the limit of the localization accuracy for a single molecule is given by lambda(em)/2pin(aroot)gammaAt, where lambda(em), n(a), gamma, A, and t denote the emission wavelength of the single molecule, the numerical aperture of the objective, the efficiency of the optical system, the emission rate of the single molecule and the acquisition time, respectively. Using Monte Carlo simulations it is shown that estimation algorithms can come close to attaining the limit given in the expression. Explicit quantitative results are also provided to show how the limit of the localization accuracy is reduced by factors such as pixelation of the detector and noise sources in the detection system. The results demonstrate what is achievable by single-molecule microscopy and provide guidelines for experimental design.
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