Regensburg 2010 – scientific programme
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BP: Fachverband Biologische Physik
BP 2: New Technologies
BP 2.4: Talk
Monday, March 22, 2010, 11:15–11:30, H43
Which parameters influence resolution in optical sub-diffraction imaging? — •Thorben Cordes1,2, Carsten Forthmann1, Christian Steinhauer1, Jan Vogelsang1, and Philip Tinnefeld1 — 1Applied Physics - Biophysics, LMU München, Amalienstr. 54, 80799 München, Germany — 2Biological Physics Research Group, Clarendon Laboratory, University of Oxford, Parks Road, Oxford, OX1 3PU, United Kingdom
In this contribution we will discuss imaging resolution of recently developed superresolution approaches from two different perspectives: (i) At first we reconsider those factors that are actually responsible for the achievable resolution limit of all superresolution techniques - either using subsequent localization of single-fluorophores (STORM, PALM)[1] or targeted readout (STED, SSIM)[1] - by incorporating the photostability of the emitter. (ii) In a second step we experimentally investigate the dependence between resolvable fluorophore density and photophysical parameters that determine the actual imaging speed in a method termed Blink-Microscopy[2]. We therefore employed single-molecule cut-and-paste[3], a method that enables molecule-by-molecule assembly of structures with features below the diffraction limit. Then complex or regular structures with patterns below the diffraction limit were used as calibration structures to characterize how parameters, such as the ON/OFF-time ratio, influence resolution and imaging speed. References: [1] S. W. Hell, Nature Methods 6 (2009) 24-31. [2] C. Steinhauer, et al. JACS 130 (2008) 16840-16841. [3] S. K. Kufer, et al. Science 319 (2008) 594-596.