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BP: Fachverband Biologische Physik

BP 31: Imaging

BP 31.3: Vortrag

Freitag, 30. März 2012, 10:15–10:30, H 1058

Cryogenic Colocalization with Nanometer Resolution — •Siegfried Weisenburger and Vahid Sandoghdar — Max Planck Institute for the Science of Light, 91058 Erlangen, Germany

The advent of super-resolution microscopy methods in the past decade caused a stir in the fluorescence microscopy community [1]. In particular, wide-field localization microscopy has pushed the resolution by more than one order of magnitude. Here, the centers of the point-spread functions of individual fluororescent molecules are determined with very high accuracy only limited by the number of collected photons [2]. At room temperature, the localization accuracy typically reaches a few tens of nanometers restricted by photobleaching. From low-temperature optical studies of single biomolecules, it is known that fluorophores are much more photostable at cryogenic temperatures [3] allowing for the detection of more photons. To overcome the accuracy limitation given by photobleaching, we introduce a new method of colocalization microscopy utilizing liquid helium temperature.

We will demonstrate our technique by colocalization measurements of a double-stranded DNA which is specifically labeled with two fluorescent molecules at a distance of ten nanometers. Furthermore, we will discuss the perspectives of this method for other biological applications.

1) S. Hell, Nat. Methods 6, 24 (2009).

2) R. Thompson et al., Biophys. J. 82, 2775 (2002).

3) R. Zodervan et al., J. Phys. Chem. A 108, 1657 (2004).