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ST: Strahlen- und Medizinphysik

ST 12: Biophysikalische Nanoskopie

ST 12.6: Talk

Wednesday, March 15, 2006, 11:40–11:50, D

Model based parameter estimation for high resolution optical microscopy — •David Baddeley, Hans Mathée, Elvira Steinwand, Claudia Batram, Christoph Cremer, and Udo Birk — Kirchhoff Institute für Physik, Universität Heidelberg

Novel methods of light microscopy such as 4Pi and Spatially Illuminated Illumination (SMI) microscopy provide us with information on a size scale significantly below that offered by conventional optical microscopy. In some cases (for instance the 2Photon 4Pi variants) high resolution imaging is possible after mathematical reconstruction, but in SMI microscopy and the one-photon 4Pi variants the Optical Transfer Function (OTF) lacks the continuous support required for unambiguous image reconstruction. The same can be true for 2Photon 4Pi microscopy in cases of imperfect alignment. In such cases a model based approach allows us to evaluate the data without the need for deconvolution. Model based approaches also offer better (sub-resolution) accuracy and reliability than image based methods, as shown in a measurement repeatability of approx 2nm in our SMI distance measurements. We present an automated, model based, approach to the analysis of SMI, 4Pi, and conventional confocal images along with various examples of its application.