Leica Science Lab - Tag : TIRF https://www.leica-microsystems.com//science-lab/tag/tirf/ Article tagged with TIRF en-US https://www.leica-microsystems.com/46588 Fluorescence Microscopy Widefield Microscopy Plant Cell Development and Morphogenesis The article discusses how tubulin molecules, which make up microtubules, in plant cells can be studied with total internal reflection fluorescence (TIRF) microscopy to better understand tubulin dynamics and cell development and morphogenesis. To visualize tubulin molecules in microtubules, an imaging solution is needed that allows them to be easily resolved near the plant cell surface. Conventional widefield microscopy detects too much out-of-focus fluorescence signal leading to reduced resolution. TIRF microscopy minimizes out-of-focus fluorescence and enables the tubulin to be clearly resolved, making TIRF a useful tool for studying molecular dynamics. https://www.leica-microsystems.com/plant-cell-development-and-morphogenesis/ Thu, 28 Jan 2021 10:59:00 +0000 PhD Olga Davydenko, PhD James DeRose https://www.leica-microsystems.com/30136 Confocal Microscopy Fluorescence Microscopy Super-Resolution CLEM TIRF Microscopy Widefield Microscopy Live-Cell Imaging Laser Microdissection Microscopy in Virology The coronavirus SARS-CoV-2, causing the Covid-19 disease effects our world in all aspects. Research to find immunization and treatment methods, in other words to fight this virus, gained highest priority in 2020. Microscopes play a significant role in this kind of research. To understand the underlaying principles of receptor binding, genome release, replication, assembly, and virus budding, as well as the response of our immune system, different methodologies and microscopes can be used. This article summarizes why microscopy is an important tool in virology and infection biology and gives examples for different microscopy technologies and their applications in these research fields. https://www.leica-microsystems.com/science-lab/microscopy-in-virology/ Mon, 13 Jul 2020 12:13:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/6282 TIRF Microscopy TIRF Publication List This monthly updated references list presents current papers using Leica AM TIRF in the major application fields for TIRF microscopy. https://www.leica-microsystems.com/science-lab/tirf-publication-list/ Tue, 03 Sep 2019 12:50:00 +0000 https://www.leica-microsystems.com/26320 Live-Cell Imaging TIRF Microscopy Where to go? Cellular Migration requires coordinated Transitions of Actin Cortex Plants, Bacteria, and Fungi possess a rigid cell wall that protects the cell and gives it shape. Animal cells, such as mammalian cells, have no outer wall, which exposes their plasma membrane to the environment. Still, they have a distinct shape, can easily alter their shape, and even change their shape to move around. This is possible thanks to a flexible “inner wall” that is composed of the actin cortex and the cytoskeleton. https://www.leica-microsystems.com/science-lab/where-to-go-cellular-migration-requires-coordinated-transitions-of-actin-cortex/ Mon, 19 Aug 2019 13:27:00 +0000 PhD Olga Davydenko, Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/18921 Super-Resolution The Molecular Architecture of Hemidesmosomes as Revealed by Super-Resolution Microscopy Hemidesmosomes have been extensively studied by immunofluorescence microscopy, but due to its limited resolution, their precise organization remained poorly understood. We studied hemidesmosome organization in cultured keratinocytes by 2- and 3-color super-resolution microscopy. We observed that in the cell periphery, nascent hemidesmosomes are associated with individual keratin filaments and that β4 is distributed along rather than under keratin filaments. By applying innovative methods to quantify molecular distances, we demonstrate that the hemidesmosomal plaque protein plectin interacts simultaneously and asymmetrically with β4 and keratin. https://www.leica-microsystems.com/science-lab/the-molecular-architecture-of-hemidesmosomes-as-revealed-by-super-resolution-microscopy/ Fri, 10 Feb 2017 09:11:00 +0000 MSc Leila Nahidiazar https://www.leica-microsystems.com/18917 Super-Resolution TIRF Microscopy Co-Orientation: Quantifying Simultaneous Co-Localization and Orientational Alignment of Filaments in Light Microscopy Co-localization analysis is a widely used tool to seek evidence for functional interactions between molecules in different color channels in microscopic images. Here we extend the basic co-localization analysis by including the orientations of the structures on which the molecules reside. We refer to the combination of co-localization of molecules and orientational alignment of the structures on which they reside as co-orientation. Because the orientation varies with the length scale at which it is evaluated, we consider this scale as a separate informative dimension in the analysis. Additionally we introduce a data driven method for testing the statistical significance of the co-orientation and provide a method for visualizing the local co-orientation strength in images. We demonstrate our methods on simulated localization microscopy data of filamentous structures, as well as experimental images of similar structures acquired with localization microscopy in different color channels. https://www.leica-microsystems.com/science-lab/co-orientation-quantifying-simultaneous-co-localization-and-orientational-alignment-of-filaments-in-light-microscopy/ Wed, 16 Sep 2015 15:20:00 +0000 Robert Nieuwenhuizen, MSc Leila Nahidiazar https://www.leica-microsystems.com/13946 TIRF Microscopy Video Talk by Daniel Axelrod: Total Internal Reflection Fluorescence (TIRF) Microscopy Total Internal Reflection Fluorescence (TIRF) Microscopy is a technique that only illuminates dye molecules near a surface. In this video, the pioneer of TIRF Microscopy describes what this technique is used for, explains the principles of the evanescent wave, gives many examples of different microscope configurations used in TIRF, and shows how polarized light TIRF can be used to image membrane orientation. https://www.leica-microsystems.com/science-lab/video-talk-by-daniel-axelrod-total-internal-reflection-fluorescence-tirf-microscopy/ Mon, 09 Mar 2015 08:45:00 +0000 https://www.leica-microsystems.com/15111 Super-Resolution Universal PAINT – Dynamic Super-Resolution Microscopy Super-resolution microscopy techniques have revolutionized biology for the last ten years. With their help cellular components can now be visualized at the size of a protein. Nevertheless, imaging living cells is a challenge for most of the super-resolution principles. https://www.leica-microsystems.com/science-lab/universal-paint-dynamic-super-resolution-microscopy/ Mon, 02 Mar 2015 08:24:00 +0000 Ph.D. Eric Hosy, Dr. Christoph Greb https://www.leica-microsystems.com/14389 Super-Resolution TIRF Microscopy Video Interview with William Hughes William Hughes works at the Garvan Institute of Medical Research, Sydney (Australia). In his Lab Head position he is interested in the causes of diabetes particularly looking at changes in exocytic behavior of pancreatic beta cells as well as fat and muscle cells. TIRF microscopy is predestined for researchers looking at cellular processes near the cytoplasmic membrane. https://www.leica-microsystems.com/science-lab/video-interview-with-william-hughes/ Wed, 24 Sep 2014 15:14:00 +0000 Dr. William Hughes, Dipl. oec.-troph. Anja Schué, Dr. Christoph Greb https://www.leica-microsystems.com/11693 Confocal Microscopy Live-Cell Imaging Super-Resolution Neuroscience Fluorescence Microscopy Live-Cell Imaging Evolves to Find New Niches Since its introduction in the 1600s, improvements in microscope technology have continually broadened the types of cells and cellular processes that can be studied. Advances in automation have made this already-simple tool faster and more capable, and time-lapse imaging reveals function and dynamics in addition to structure. Live-cell imaging has enabled us to witness incredible moments in biology in unprecedented detail. Even embryogenesis – the process of cell division and cellular differentiation that occurs at the earliest stages of life – has recently been captured. https://www.leica-microsystems.com/science-lab/live-cell-imaging-evolves-to-find-new-niches/ Mon, 25 Nov 2013 10:15:00 +0000 https://www.leica-microsystems.com/5945 TIRF Microscopy Controlling the TIRF Penetration Depth is Mandatory for Reproducible Results The main feature of total internal reflection fluorescence (TIRF) microscopy is the employment of an evanescent wave for the excitation of fluorophores instead of using direct light. A property of the evanescent wave, which arises from the glass/water or glass/specimen interface, is that its propagation in z-direction gradually degrades, limiting its penetration depth into the specimen to some hundred nanometers. https://www.leica-microsystems.com/science-lab/controlling-the-tirf-penetration-depth-is-mandatory-for-reproducible-results/ Tue, 24 Apr 2012 22:00:00 +0000 Dr. Thomas Veitinger https://www.leica-microsystems.com/5403 TIRF Microscopy Live-Cell Imaging Widefield Microscopy Total Internal Reflection Fluorescence (TIRF) Microscopy Total internal reflection fluorescence (TIRF) is a special technique in fluorescence microscopy developed by Daniel Axelrod at the University of Michigan, Ann Arbor in the early 1980s. TIRF microscopy delivers images with an outstandingly high axial resolution below 100 nm. This allows the observation of membrane-associated processes. https://www.leica-microsystems.com/science-lab/total-internal-reflection-fluorescence-tirf-microscopy/ Sun, 11 Mar 2012 23:00:00 +0000 Wymke Ockenga https://www.leica-microsystems.com/5405 Live-Cell Imaging TIRF Microscopy Widefield Microscopy Applications of TIRF Microscopy in Life Science Research The special feature of TIRF microscopy is the employment of an evanescent field for fluorophore excitation. Unlike standard widefield fluorescence illumination procedures with arc lamps, LEDs or lasers, the evanescent field only penetrates the specimen by about 100 nm starting from the coverslip/medium interface. https://www.leica-microsystems.com/science-lab/applications-of-tirf-microscopy-in-life-science-research/ Sun, 11 Mar 2012 23:00:00 +0000 Wymke Ockenga, Dr. Thomas Veitinger https://www.leica-microsystems.com/5175 TIRF Microscopy Live-Cell Imaging TIRF Microscopy Explores Cellular Transport Processes Due to their special role in organ function and the exchange of biological components some body cells developed certain polarization characteristics. These are reflected in differences of their plasma membrane composition. The essential and fascinating task of polarized protein transport in epithelial cells is to get the right protein into the right membrane. https://www.leica-microsystems.com/science-lab/protein-transport-processes-at-the-apical-membrane-of-polarized-epithelial-cells/ Sun, 11 Mar 2012 23:00:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/5108 Confocal Microscopy Live-Cell Imaging Live-cell Imaging Techniques The understanding of complex and/or fast cellular dynamics is an important step for exploring biological processes. Therefore, today’s life science research is increasingly focusing on dynamic processes like cell migration, morphological changes of cells, organs or whole animals and physiological (e.g. changes of intracellular ion composition) events in living specimens in real time. https://www.leica-microsystems.com/science-lab/live-cell-imaging-techniques/ Thu, 23 Feb 2012 23:00:00 +0000 Dr. Thomas Veitinger, Dr. Zhongxiang Jiang https://www.leica-microsystems.com/2994 Live-Cell Imaging TIRF Microscopy TIRF Microscopy of the Apical Membrane of Polarized Epithelial Cells Application of TIRF microscopy (Total Internal Reflection Fluorescence) allows the visualization of structures at the apical surface of polarized epithelial cells that have been hidden in conventional fluorescence microscopy images. Hence, the approach reveals new insights into the composition of this characteristic cell pole that elucidate processes in apical protein trafficking. https://www.leica-microsystems.com/science-lab/tirf-microscopy-of-the-apical-membrane-of-polarized-epithelial-cells/ Tue, 12 Apr 2011 22:00:00 +0000 Dr. Christoph Greb, Prof. Ralf Jacob https://www.leica-microsystems.com/4411 Super-Resolution A Guide to Super-Resolution Fluorescence Microscopy For centuries, cell biology has been based on light microscopy and at the same time been limited by its optical resolution. However, several new technologies have been developed recently that bypass this limit. https://www.leica-microsystems.com/science-lab/a-guide-to-super-resolution-fluorescence-microscopy/ Sun, 18 Jul 2010 22:00:00 +0000 https://www.leica-microsystems.com/3006 Live-Cell Imaging TIRF Microscopy Exploring Cell Logistics Using TIRF microscopy, scientists have been able to take a closer look at intracellular transport processes with the example of the galactose-binding protein Galectin-3, which has been identified as a potential apical sorting receptor. https://www.leica-microsystems.com/science-lab/exploring-cell-logistics/ Fri, 02 Nov 2007 11:29:00 +0000 Prof. Ralf Jacob, Dipl. Biol. Dominik Schneider