Structured illumination microscopy using photoswitchable fluorescent proteins

Liisa Hirvonen; Ondrej Mandula; Kai Wicker; Rainer Heintzmann

doi:10.1117/12.763021

12 February 2008 Structured illumination microscopy using photoswitchable fluorescent proteins

Liisa Hirvonen, Ondrej Mandula, Kai Wicker, Rainer Heintzmann

Proceedings Volume 6861, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV; 68610L (2008) https://doi.org/10.1117/12.763021
Event: SPIE BiOS, 2008, San Jose, California, United States

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.

Citation Download Citation

Liisa Hirvonen, Ondrej Mandula, Kai Wicker, and Rainer Heintzmann "Structured illumination microscopy using photoswitchable fluorescent proteins", Proc. SPIE 6861, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV, 68610L (12 February 2008); https://doi.org/10.1117/12.763021

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