Leica Science Lab - Tag : FCS https://www.leica-microsystems.com//science-lab/tag/tags/fcs/show/Tag/ Article tagged with FCS en-US https://www.leica-microsystems.com/20133 Quantitative Fluorescence FCS - Fluorescence Correlation Spectroscopy FCS is a fluorescence-based measurement method. Fluorescent molecules passing through a strongly focused, fixed laser beam are excited for fluorescence emission. After passing a confocal pinhole, the emitted photons are registered using very sensitive detectors. https://www.leica-microsystems.com//science-lab/fcs-fluorescence-correlation-spectroscopy/ Wed, 28 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20131 Quantitative Fluorescence FLCS - Fluorescence Lifetime Correlation Spectroscopy Essentially, FCS can be performed with a continuous-wave laser. Using pulsed lasers allows even more sophisticated analysis possibilities, such as time-gated FCS or Fluorescence Lifetime Correlation Spectroscopy (FLCS). Both methods make use of the additional information obtained by the simultaneous measurement of the fluorescence lifetime. https://www.leica-microsystems.com//science-lab/flcs-fluorescence-lifetime-correlation-spectroscopy/ Mon, 26 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/20132 Quantitative Fluorescence FCCS - Fluorescence Cross-Correlation Spectroscopy FCCS (Fluorescence Cross-Correlation Spectroscopy) can be measured using the Leica TCS SP8 FCS system. Similar to FCS , it analyzes fluorescence intensity fluctuations derived from a small observation volume. https://www.leica-microsystems.com//science-lab/fccs-fluorescence-cross-correlation-spectroscopy/ Sun, 25 Mar 2018 22:00:00 +0000 Dr. Lioba Kuschel https://www.leica-microsystems.com/18278 Super-Resolution Live-Cell Imaging Super-Resolution Optical Microscopy of Lipid Plasma Membrane Dynamics Plasma membrane dynamics are an important ruler of cellular activity, particularly through the interaction and diffusion dynamics of membrane-embedded proteins and lipids. FCS (fluorescence correlation spectroscopy) on an optical (confocal) microscope is a popular tool for investigating such dynamics. Unfortunately, its full applicability is constrained by the limited spatial resolution of a conventional optical microscope. The present chapter depicts the combination of optical super-resolution STED (stimulated emission depletion) microscopy with FCS, and why it is an important tool for investigating molecular membrane dynamics in living cells. Compared with conventional FCS, the STED-FCS approach demonstrates an improved possibility to distinguish free from anomalous molecular diffusion, and thus to give new insights into lipid–protein interactions and the traditional lipid ‘raft’ theory. https://www.leica-microsystems.com//science-lab/super-resolution-optical-microscopy-of-lipid-plasma-membrane-dynamics/ Fri, 23 Dec 2016 14:16:00 +0000 Prof. Christian Eggeling https://www.leica-microsystems.com/18271 Super-Resolution Quantitative Fluorescence Live-Cell Imaging STED-FLCS: An Advanced Tool to Reveal Spatiotemporal Heterogeneity of Molecular Membrane Dynamics Heterogeneous diffusion dynamics of molecules play an important role in many cellular signaling events, such as of lipids in plasma membrane bioactivity. However, these dynamics can often only be visualized by single-molecule and super-resolution optical microscopy techniques. Using fluorescence lifetime correlation spectroscopy (FLCS, an extension of fluorescence correlation spectroscopy, FCS) on a super-resolution stimulated emission depletion (STED) microscope, we here extend previous observations of nanoscale lipid dynamics in the plasma membrane of living mammalian cells. https://www.leica-microsystems.com//science-lab/sted-flcs-an-advanced-tool-to-reveal-spatiotemporal-heterogeneity-of-molecular-membrane-dynamics/ Mon, 11 Jul 2016 08:53:00 +0000 Ph.D. Giuseppe Vicidomini https://www.leica-microsystems.com/18431 Quantitative Fluorescence Live-Cell Imaging Individual Macromolecule Motion in a Crowded Living Cell There is solid evidence for analyzing fluorescence correlation and dual color fluorescence crosscorrelation spectroscopy data (FCS and dual color FCCS) in cellular applications by equations based on anomalous subdiffusion. Using equations based on normal diffusion causes artifacts of the fitted biological system response parameters and of the interpretations of the FCS and dual color FCCS data in the crowded environment of living cells. Equations based on normal diffusion are not valid in living cells. The original article embraces the status of the experimental situation and touches obstacles that still hinder the applications of single molecules in the cellular environment. https://www.leica-microsystems.com//science-lab/individual-macromolecule-motion-in-a-crowded-living-cell/ Mon, 27 Jun 2016 04:57:00 +0000 https://www.leica-microsystems.com/16176 Super-Resolution Quantitative Fluorescence A Straightforward Approach for Gated STED-FCS to Investigate Lipid Membrane Dynamics Recent years have seen the development of multiple technologies to investigate, with great spatial and temporal resolution, the dynamics of lipids in cellular and model membranes. One of these approaches is the combination of far-field super-resolution stimulated-emission-depletion (STED) microscopy with fluorescence correlation spectroscopy (FCS). STED-FCS combines the diffraction-unlimited spatial resolution of STED microscopy with the statistical accuracy of FCS to determine sub-millisecond-fast molecular dynamics with single-molecule sensitivity. https://www.leica-microsystems.com//science-lab/a-straightforward-approach-for-gated-sted-fcs-to-investigate-lipid-membrane-dynamics/ Mon, 03 Aug 2015 10:29:00 +0000 https://www.leica-microsystems.com/16174 Super-Resolution Quantitative Fluorescence Live-Cell Imaging Cortical Actin Networks Induce Spatio-temporal Confinement of Phospholipids in the Plasma Membrane – A Minimally Invasive Investigation by STED-FCS Important discoveries in the last decades have changed our view of the plasma membrane organisation. Specifically, the cortical cytoskeleton has emerged as a key modulator of the lateral diffusion of membrane proteins. Cytoskeleton-dependent compartmentalised lipid diffusion has been proposed, but this concept remains controversial because this phenomenon has thus far only been observed with artefact-prone probes in combination with a single technique: single particle tracking. https://www.leica-microsystems.com//science-lab/cortical-actin-networks-induce-spatio-temporal-confinement-of-phospholipids-in-the-plasma-membrane-a-minimally-invasive-investigation-by-sted-fcs/ Fri, 24 Jul 2015 13:02:00 +0000 Débora Machado Andrade https://www.leica-microsystems.com/15239 Super-Resolution Quantitative Fluorescence A Lipid Bound Actin Meshwork Organizes Liquid Phase Separation in Model Membranes The eukaryotic cell membrane is connected to a dense actin rich cortex. We present FCS and STED experiments showing that dense membrane bound actin networks have severe influence on lipid phase separation. Our results reveal a mechanism how cells may prevent macroscopic demixing of their membrane components, while at the same time regulate the local membrane composition. https://www.leica-microsystems.com//science-lab/a-lipid-bound-actin-meshwork-organizes-liquid-phase-separation-in-model-membranes/ Wed, 15 Jul 2015 13:49:00 +0000 https://www.leica-microsystems.com/15347 Super-Resolution STED Nanoscopy: A Glimpse into the Future The well-known saying of "Seeing is believing" became even more apt in biology when stimulated emission depletion (STED) nanoscopy was introduced in 1994 by the Nobel laureate S. Hell and coworkers. This article gives an overview of the various cutting-edge implementations of STED nanoscopy and tries to shine a light into the future: imaging everything faster with unprecedented sensitivity and label-free. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-a-glimpse-into-the-future/ Mon, 23 Mar 2015 18:08:00 +0000 Paolo Bianchini https://www.leica-microsystems.com/14017 Super-Resolution Image Restoration and Deconvolution Abstracts of the 4th European Super-Resolution User-Club Meeting The 4th Super-Resolution User Club Meeting was held in collaboration with Christian Eggeling and the Weatherall Institute of Molecular Medicine in Oxford, UK. Here we present the abstracts of the talks and interviews with participants. https://www.leica-microsystems.com//science-lab/abstracts-of-the-4th-european-super-resolution-user-club-meeting/ Fri, 08 Aug 2014 08:09:00 +0000 Prof. Christian Eggeling, Dr. Timo Zimmermann, Dr. Katrin Willig, PhD Kees Jalink, Dr. Marko Lampe, Prof. Mark Neil, Dr. Sergi Padilla-Parra, MSc Leila Nahidiazar, Ph.D. Giuseppe Vicidomini, Ph.D. Marc van Zandvoort https://www.leica-microsystems.com/10135 Super-Resolution Widefield Microscopy Image Restoration and Deconvolution Abstracts of the 3rd European Super-Resolution User-Club Meeting The 3rd meeting of the Leica Super-Resolution User Club was held from June 17th to 19th, 2013 in collaboration with Alberto Diaspro and the Italian Institute of Technology (IIT) in Genoa. Confocal and widefield super-resolution users from ten European countries took three days’ out to deepen their knowledge on super-resolution techniques and applications and make use of an opportunity for full exchange of experiences. https://www.leica-microsystems.com//science-lab/abstracts-of-the-3rd-european-super-resolution-user-club-meeting/ Mon, 08 Jul 2013 10:27:00 +0000 Prof. Alberto Diaspro, Ph.D. Illaria Testa, Prof. Silvio Rizzoli, Ph.D. Giuseppe Vicidomini, Prof. Ralf Jacob, Ph.D. Eric Hosy, Prof. Colin Sheppard, Zeno Lavagnino https://www.leica-microsystems.com/7735 Super-Resolution Widefield Microscopy Image Restoration and Deconvolution Abstracts of the 2nd European Super-Resolution User-Club Meeting The 2nd meeting of the Leica Super-resolution User club was held from September 25 to 27, 2012 in collaboration with the Science for Life Laboratory at the Karolinska Institute, Stockholm, Sweden. With a mixture of engaging talks by key experts in the field of super-resolution microscopy and stimulating discussion sessions, the meeting proved as popular as last year’s event, attracting a wide range of scientists interested in both confocal and widefield super-resolution and sample preparation techniques. https://www.leica-microsystems.com//science-lab/abstracts-of-the-2nd-european-super-resolution-user-club-meeting/ Thu, 01 Nov 2012 23:00:00 +0000 Prof. Hjalmar Brismar, Prof. Dr. Dr. h.c. Stefan Hell, Prof. Christian Eggeling, Dr. Anna Szymborska, Prof. Alberto Diaspro, Ph.D. Eric Hosy, Ph.D. Giuseppe Vicidomini, Hans van der Voort https://www.leica-microsystems.com/6609 Confocal Microscopy Quantitative Fluorescence Fluorescence Correlation Spectroscopy Fluorescence correlation spectroscopy (FCS) measures fluctuations of fluorescence intensity in a sub-femtolitre volume to detect such parameters as the diffusion time, number of molecules or dark states of fluorescently labeled molecules. The technique was independently developed by Watt Webb and Rudolf Rigler during the early 1970s. https://www.leica-microsystems.com//science-lab/fluorescence-correlation-spectroscopy/ Thu, 02 Aug 2012 22:00:00 +0000 Dr. Constantin Kappel https://www.leica-microsystems.com/5899 Quantitative Fluorescence Quantitative Fluorescence Seeing is believing – and measuring is knowing. Microscopes generate images that are not only used for illustration, but are also subject to quantification. More advanced techniques use illumination patterns (without image formation) or do not generate an image at all – but are still microscopical techniques. These F-techniques are becoming increasingly important in current biosciences. https://www.leica-microsystems.com//science-lab/quantitative-fluorescence/ Wed, 18 Apr 2012 22:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/5082 Confocal Microscopy Quantitative Fluorescence FLCS – Advances in Fluorescence Correlation Spectroscopy The characterization of substances at the single molecule level has become part of the standard repertoire of scientific research institutes. One of the most common methods is Fluorescence Correlation Spectroscopy (FCS), which can be used to examine the dynamics and concentration of fluorescent molecules in solution. https://www.leica-microsystems.com//science-lab/flcs-advances-in-fluorescence-correlation-spectroscopy/ Wed, 14 Dec 2011 23:00:00 +0000 Dr. Andreas Bülter, Dr. Andrea Bleckmann, Uwe Ortmann