Structured illumination microscopy using photoswitchable fluorescent proteins

Liisa Hirvonen, Ondrej Mandula, Kai Wicker, Rainer Heintzmann

Research output: Chapter in Book/Conference paperConference paper

18 Citations (Scopus)

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 fine 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.

Original languageEnglish
Title of host publicationThree-Dimensional and Multidimensional Microscopy
Subtitle of host publicationImage Acquisition and Processing XV
DOIs
Publication statusPublished - 21 Apr 2008
Externally publishedYes
EventThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV - San Jose, CA, United States
Duration: 21 Jan 200824 Jan 2008

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6861
ISSN (Print)1605-7422

Conference

ConferenceThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV
CountryUnited States
CitySan Jose, CA
Period21/01/0824/01/08

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