21 ноября (четверг), 1705, 115 КПМ
Dr. Klaus Weisshart
Carl Zeiss Microscopy GmbH, BioSciences Division Carl Zeiss Microscopy GmbH, BioSciences Division
Super-resolution microscopy in 3D – Photoactivated Localization and Structured Illumination Microscopy to study cellular structures and dynamics
Resolving ultrafine details of subcellular structures is key to understand the organization and functioning of cellular networks. Recent advances in far-field fluorescence microscopy provide the necessary tools to analyze these structures with resolutions well below the classical diffraction limit in all three-dimensions. These technical advances went hand in hand with improved versions of photo-switchable fluorophores that allowed to push resolution limits further down. The wide spectrum of suitable dyes along with the high contrast achieved endow these fluorescence based superresolution (SR) techniques with the power to study the complexity of sub-cellular organelles and the relation of their constituting components down to the molecular level and under physiological conditions. They provide us in this way with a far better understanding of the assembly of macromolecular complexes and their functions within a cell than has been possible before employing conventional imaging methods. Here we will give an overview of the technical state-of-the art of two of these technologies, Structured Illumination Microscopy (SIM) and Photoactivated Localization Microscopy (PALM), and provide typical application examples in this exciting field.
25 ноября (понедельник), 1705, 110 КПМ
Dr. Thomas Gensch
ICS-4, Research Centre J?lich Super-resolution
Fluorescence Microscopy of Cellular Components
Fluorescence Microscopy has become an indispensable tool in cell biology being powerful in visualizing the processes of life in fixed and living cells. But - like all optical microscopy methods - it suffers from the diffraction limited resolution that is for visible light in the range of 200 - 300 nm. Since the days of Ernst Abbe this limitation was widely accepted until at the end of the last millennium several experimental approaches – like stimulated emission depletion (STED), structured illumination microscopy (SIM) and single molecule localization microscopies (SMLM, PALM, STORM) - had been suggested and were realized in the first decade of the new millennium. These methods are subsumed under the name super-resolution fluorescence microscopy. Since their first realizations super-resolution fluorescence microscopy has tremendously developed and allows nowadays images of biological cells with spatial and time resolutions of better than 50 nm and down to 10s. The lecture will give an overview about the different techniques, some applications on biological systems from the literature and the super-resolution fluorescence microscopy in our own laboratory. We started in 2009 to build a widefield/TIRF fluorescence microscope suitable for localization-based super-resolution microscopy. We study with PALM and directSTORM several cellular processes including actin filament structure and dynamics, mitochondrial protein import, proteins in synapses and synaptic vesicles of hippocampal neurons and organization and trafficking of ion channels and transporter proteins.