Berlin 2015 – scientific programme
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CPP: Fachverband Chemische Physik und Polymerphysik
CPP 28: New Instruments and Methods
CPP 28.7: Talk
Tuesday, March 17, 2015, 15:30–15:45, C 264
Resolution Enhancement For Low-Temperature Scanning Microscopy By Cryostat Immersion Imaging — •Michael Metzger1, Alexander Konrad1, Alfred J. Meixner1, and Marc Brecht2 — 1Institute of Physical and Theoretical Chemistry, University of Tuebingen, Germany — 2Zurich University of Applied Science, Institute of Applied Mathematics and Physics, Winterthur, Switzerland
One convenient way to increase the resolution of e.g. fluorescence images is realized by confocal microscopy with an objective of high numerical aperture and immersion oil. The combination of immersion fluids with high-performance objectives is however exceedingly problematic under low temperature conditions. A new construction of a scanning stage and a well-chosen immersion fluid enables us to immerse an objective together with the sample positioned inside a cryostat at cryogenic temperatures. Heating the sample chamber over the melting point of an appropriate chosen immersion fluid (e.g. 1-propanol) allows us to move the objective into the melted immersion droplet and thereby to increase the refractive index between the objective lens and the sample. We recorded confocal fluorescence images of quantum dots at 160 K with a high NA objective. By determining the point spread function of imaged single quantum dots the effective numerical aperture was appointed to be larger than unity (1.08). The presented method provides new opportunities e.g. for studies on biological systems like vitrified cells at low temperature and is also of relevance for correlative light and electron cryo microscopy.