Month: August 2013

To correlate dynamic events in adherent cells with both ultrastructural and 3D information, we developed a method for cultured cells that combines confocal time-lapse images of GFP-tagged proteins with electron microscopy. With laser micro-patterned culture substrate, we created coordinates that were conserved at every step of the sample preparation and visualization processes. Specifically designed for …

From Dynamic Live Cell Imaging to 3D Ultrastructure: Novel Integrated Methods for High Pressure Freezing and Correlative Light-Electron Microscopy Read More »

We developed a new class of two-photon excitation–stimulated emission depletion (2PE–STED) optical microscope. In this work, we show the opportunity to perform superresolved fluorescence imaging, exciting and stimulating the emission of a fluorophore by means of a single wavelength. We show that a widely used red-emitting fluorophore, ATTO647N, can be two-photon excited at a wavelength …

Single-Wavelength Two-Photon Excitation-stimulated Emission Depletion (SW2PE-STED) Superresolution Imaging Read More »

The new method combines two types of microscopy. Using a confocal scanning microscope, fluorescence images are recorded point by point at fixed time intervals. Hence, the images contain an implicit time structure. This information can be used with the help of raster image correlation spectroscopy (RICS) to determine the dynamics of biomolecules, such as proteins, …

Watching Molecule Movements in Live Cells Read More »

Fluorescence microscopy and other light-based applications require optical filters that have demanding spectral and physical characteristics. Often, these characteristics are application-specific and an optic that might be appropriate and optimal for one is both inappropriate and sub-optimal for another. The Handbook of Optical Filters for Fluorescence Microscopy is a compilation of the principles that the …

Handbook of Optical Filters for Fluorescence Microscopy Read More »

Silvio Rizzoli is one the very first scientists to have worked with STED microscopes. Asked about his personal highlights of his work with STED microscopy in the past years, he mentions three milestones: In the first biological application of STED microscopy (Willig et al. [2006]), Rizzoli, Stefan Hell and colleagues from Göttingen used STED to …

New Labeling Tools Can Help to Realize the Full Potential of Super-Resolution Microscopy Read More »