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Super-resolution techniques like PALM and STORM require accurate localization of single fluorophores detected using a CCD. Popular localization algorithms inefficiently assume each photon registered by a pixel can only come from an area in the specimen corresponding to that pixel (not from neighboring areas), before iteratively (slowly) fitting a Gaussian to pixel intensity; they fail with noisy images. We present an alternative; a probability distribution extending over many pixels is assigned to each photon, and independent distributions are joined to describe emitter location. We compare algorithms, and recommend which serves best under different conditions. At low signal-to-noise ratios, ours is 2-fold more precise than others, and 2 orders of magnitude faster; at high ratios, it closely approximates the maximum likelihood estimate.

Type

Journal article

Journal

Opt Express

Publication Date

30/07/2012

Volume

20

Pages

18478 - 18493

Keywords

Animals, COS Cells, Cercopithecus aethiops, Image Processing, Computer-Assisted, Limit of Detection, Microtubules, Optical Phenomena