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Proceedings Paper

Structured illumination microscopy using photoswitchable fluorescent proteins
Author(s): Liisa Hirvonen; Ondrej Mandula; Kai Wicker; Rainer Heintzmann
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Paper Abstract

In fluorescence microscopy the lateral resolution is limited to about 200 nm because of diffraction. Resolution improvement by a factor of two can be achieved using structured illumination, where a ine grating is projected onto the sample, and the final image is reconstructed from a set of images taken at different grating positions. Further resolution improvement can be achieved by saturating the transitions involved in fluorescence emission. Recently discovered photoswitchable proteins undergo transitions that are saturable at low illumination intensity. Combining this concept with structured illumination, theoretically unlimited resolution can be achieved, where the smallest resolvable distance will be determined by signal-to-noise ratio. This work focuses on the use of the photoswitchable protein Dronpa with structured illumination to achieve nanometre scale resolution in fixed cells.

Paper Details

Date Published: 12 February 2008
PDF: 8 pages
Proc. SPIE 6861, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV, 68610L (12 February 2008); doi: 10.1117/12.763021
Show Author Affiliations
Liisa Hirvonen, King's College London (United Kingdom)
Ondrej Mandula, King's College London (United Kingdom)
Kai Wicker, King's College London (United Kingdom)
Rainer Heintzmann, King's College London (United Kingdom)

Published in SPIE Proceedings Vol. 6861:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson; Thomas G. Brown, Editor(s)

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