Open AccessLow power super resolution fluorescence microscopy by lifetime modification and image reconstruction
Author(s) -
Richard J. Marsh,
Siân Culley,
Angus J. Bain
Publication year - 2014
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.22.012327
Subject(s) - optics , microscopy , point spread function , fluorescence , materials science , resolution (logic) , fluorescence microscope , sted microscopy , diffraction , image resolution , super resolution microscopy , stimulated emission , physics , laser , computer science , artificial intelligence
We demonstrate a new method for obtaining sub-diffraction resolution in fluorescence microscopy. The technique involves the analysis of the time evolution of fluorescence images in the presence of weak and unstructured (fundamental Gaussian) continuous wave stimulated emission depletion. A reduced point spread functions (PSF) is obtained by the recombination of time segments of the evolving image. A significant reduction in the PSF for 20 nm fluorescent beads (ca. 240 nm to 125 nm) is obtained with an on-sample power of 7.5 mW (17 MW/cm2) - substantially lower than that required for spatially structured stimulated emission depletion microscopy.