From millimeters to nanometers - reducing the scale in microscopy
Herbert Schneckenburger and Verena Richter
Institute of Applied Research, Aalen University, Beethovenstr. 1, 73430 Aalen, Germany
Abstract
While conventional wide-field microscopy is limited in resolution, contrast and depth of focus, novel methods of super-resolution and 3D imaging overcome these limitations. However, high light exposure with a risk of phototoxicity to living cells and organisms has to be avoided. Therefore, we developed super-resolution methods, which are more tolerable to living cells, in particular SIM in combination with Total Internal reflection Fluorescence Microscopy (TIRFM) or SIM in combination with Axial Tomography. In addition, variable-angle TIRFM permitted measurements of cell membrane topology with nanometer resolution, e.g. for studies of cell growth or interaction with cytotoxic agents. In addition, molecular parameters in the nanometer range can be deduced from spectral data, fluorescence lifetimes or non-radiative energy transfer (FRET) between a donor and an acceptor molecule. Since this interaction may be sensitive to stimulation by pharmaceutical agents, methods (e.g. TIRFM) are transferred from a fluorescence microscope to a multi-well reader system for pharmacological tests and simultaneous detection of large cell populations.
Speaker
Herbert Schneckenburger
Institute of Applied Research, Aalen University
Germany
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