Month: March 2012

Total Internal Reflection Fluorescence (TIRF) Microscopy

There are two approaches for achieving total internal reflection in optics: one is prism-based and the other objective-based. In prism-based TIRF microscopy, a prism is attached to the coverslip’s surface which directs a focused light beam or laser towards the coverslip/medium interface. With the help of the prism the angle of the penetrating light is …

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Applications of TIRF Microscopy in Life Science Research

By employing a spatially restricted evanescent field for the excitation of fluorophores, TIRF microscopy allows observation of the localization and dynamics of molecules and processes in an optical section near the plasma membrane (usually around 100 nm). This is advantageous for many applications dealing with processes in or close to the plasma membrane. TIRF microscopy is …

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Protein Transport Processes at the Apical Membrane of Polarized Epithelial Cells

Higher organisms such as mammals are composed of a number of organs that all fulfill specific tasks for which they need to communicate. To enable targeted, controlled communication, these functional units have to be separated, both spatially and biochemically. All animal organs are therefore surrounded by epithelia which, besides protecting them from harmful environmental influences, …

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Visualization of the Natural Killer Cell Immune Synapse by Super-Resolution Nanoscopy

In addition to our use of super-resolution in studies described above, we continue to focus on the interaction of lytic granules with the cytoskeleton. Using STED nanoscopy we visualized individual myosin IIa filaments wrapped around granules, both isolated and within whole cells [8]. These images enabled us to discern between alternative models of myosin IIa-granule …

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Dual Channel STED Nanoscopy of Lytic Granules on Actin Filaments in Natural Killer Cells

Natural killer (NK) cells are innate immune effectors that eliminate diseased and tumorigenic targets through the directed secretion of specialized secretory lysosomes, termed lytic granules. This directed secretion is triggered following the formation of an immunological synapse (IS), which is characterized by actin re-modeling and receptor organization at the interface between the NK cell and …

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