2011/01/28 by Stefan Geissbuehler, Claudio Dellagiacoma, Theo Lasser · 1 citation
Biochemistry, Genetics and Molecular Biology · Physics and Astronomy · #Advanced Fluorescence Microscopy Techniques #Cell Image Analysis Techniques #Advanced Optical Sensing Technologies
paper · doi:10.1364/boe.2.000408
openalex publication_date 2011/01/28 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/02
A straightforward method to achieve super-resolution consists of taking an image sequence of stochastically blinking emitters using a standard wide-field fluorescence microscope. Densely packed single molecules can be distinguished sequentially in time using high-precision localization algorithms (e.g., PALM and STORM) or by analyzing the statistics of the temporal fluctuations (SOFI). In a face-to-face comparison of the two post-processing algorithms, we show that localization-based super-resolution can deliver higher resolution enhancements but imposes significant constraints on the blinking behavior of the probes, which limits its applicability for live-cell imaging. SOFI, on the other hand, works more consistently over different photo-switching kinetics and also delivers information about the specific blinking statistics. Its suitability for low SNR acquisition reveals SOFI's potential as a high-speed super-resolution imaging technique.