Localization of a fluorescent source without numerical fitting.
We present an algebraic solution to the problem of localizing a single fluorescent particle with sub-diffraction-limit accuracy. The algorithm is derived and its performance studied experimentally. Isolated 20 nm fluorescent beads were imaged using a wide-field microscope at two different positions separated by 100 nm and at a range of signal-to-noise ratios (SNR). The data were analyzed using both the new algorithm and the standard approach of fitting the data to a Gaussian profile. Results indicate that the proposed approach is nearly as accurate as Gaussian fitting across a wide range of SNR while executing over 200 times faster. In addition, the new algorithm is able to localize at lower SNR than the fitting method.
Andersson
Sean B
SB
Department of Mechanical Engineering, Boston University, Boston, MA 02215, USA. sanderss@bu.edu
D000465:Algorithms; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D053758:Nanoparticles / Q000648:ultrastructure*; D010363:Pattern Recognition, Automated / Q000379:methods*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*