Jochen Sieber

Confocal imaging at the nanoscale with two-color STED microscopy

STED microscopy enables confocal imaging of biological samples with a resolution that is not limited by diffraction. It provides new insights in various fields of biology, such as membrane biology, neurobiology and physiology. Its three dimensional sectioning ability allows the acquisition of high resolution image stacks. Furthermore, STED microscopy enables the recording of dynamic processes and live cell images. We present two-color imaging in confocal STED microscopy with a single STED wavelength. Pulsed and continuous wave lasers in the visible and near infra-red wavelengths range are used for stimulated emission. The resolution enhancement is demonstrated in comparison to confocal imaging with biological specimens.

New developments in STED Microscopy

STED microscopy has gained recognition as a method to break the diffraction limit of conventional light microscopy. Despite being a new technique, STED is already successfully implemented in life science research. The resolution enhancement is achieved by depleting fluorescent markers via stimulated emission. The performance is significantly dependent on the laser source and the fluorescence markers. Therefore the use of novel fluorescent markers in conjunction with the right laser system was the main focus of our research. We present new developments and applications of STED microscopy, unraveling structural details on scales below 90nm and give an overview of required specifications for the solid state laser systems.