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Open Access Original article

Optical photon reassignment microscopy (OPRA)

Stephan Roth12, Colin JR Sheppard3, Kai Wicker12 and Rainer Heintzmann124*

Author Affiliations

1 Institute of Photonic Technology, Albert-Einstein-Str.9, 07745, Jena, Germany

2 Institute of Physical Chemistry, Abbe Center of Photonics, Friedrich-Schiller-University Jena, Helmholtzweg 4, 07743, Jena, Germany

3 Nanophysics, Instituto Italiano di Tecnologia, via Morego 30, 16163, Genoa, Italy

4 King’s College London, Randall Division, NHH, SE1 1UL, London, UK

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Optical Nanoscopy 2013, 2:5  doi:10.1186/2192-2853-2-5

Published: 18 October 2013

Abstract

To enhance the resolution of a confocal laser scanning microscope the additional information of a pinhole plane image taken at every excitation scan position can be used (Sheppard 1988). This photon reassignment principle is based on the fact that the most probable position of an emitter is at half way between the nominal focus of the excitation laser and the position corresponding to the (off centre) detection position. Therefore, by reassigning the detected photons to this place, an image with enhanced detection efficiency and resolution is obtained. Here we present optical photon reassignment microscopy (OPRA) which realizes this concept in an all-optical way obviating the need for image-processing. With the help of an additional intermediate optical beam expansion between descanning and a further rescanning of the detected light, an image with the advantages of photon reassignment can be acquired. However, just as in computational photon reassignment, a loss in confocal sectioning performance is caused by working with relatively open pinholes. The OPRA system shares properties such as flexibility and ease of use with a confocal laser scanning microscope, and is therefore expected to be of use for future biomedical routine research.

Keywords:
Photon reassignment; Image scanning microscopy; Confocal laser scanning microscopy