Leica Science Lab - Tag : F-Techniques https://www.leica-microsystems.com//science-lab/tag/f-techniques/ Article tagged with F-Techniques en-US https://www.leica-microsystems.com/44422 Confocal Microscopy Live-Cell Imaging Advanced Quantitative Fluorescence Microscopy to Probe the Molecular Dynamics of Viral Entry Viral entry into the host cell requires the coordination of many cellular and viral proteins in a precise order. Modern microscopy techniques are now allowing researchers to investigate these interactions with higher spatiotemporal resolution than ever before. Here we present two examples from the field of HIV research that make use of an innovative quantitative imaging approach as well as cutting edge fluorescence lifetime-based confocal microscopy methods to gain novel insights into how HIV fuses to cell membranes and enters the cell. https://www.leica-microsystems.com/advanced-quantitative-fluorescence-microscopy-to-probe-the-molecular-dynamics-of-viral-entry/ Wed, 11 Nov 2020 09:48:00 +0000 PhD Ben Libberton https://www.leica-microsystems.com/13262 Quantitative Imaging FRAP with TCS SP8 Resonant Scanner Fast FRAP experiments need a sufficient number of measurement points for meaningful interpretation and fitting analysis. To study very fast translocational processes, the use of a resonant scanner (RS) is preferred. The advantage in using FRAP with the RS is that statistics are much better in experiments that require fast acquisition: If the half time of recovery is about 0.5 sec you may have only about 3 to 4 data points using the conventional scanner, whereas with the resonant scanner you can get about 20 data points. https://www.leica-microsystems.com/science-lab/frap-with-tcs-sp8-resonant-scanner/ Tue, 26 Jan 2016 15:39:00 +0000 Dr. Jan Schröder https://www.leica-microsystems.com/9759 Quantitative Imaging Step by Step Guide for FRAP Experiments Fluorescence Recovery After Photobleaching (FRAP) has been considered the most widely applied method for observing translational diffusion processes of macromolecules. The resulting information can be used to determine kinetic properties, like the diffusion coefficient, mobile fraction, and transport rate of the fluorescently labeled molecules. FRAP employs irradiation of a fluorophore with a short laser pulse. State of the art laser scanning microscopes, like the TCS SP8 confocal system, have the advantage of using a high intensity laser for photobleaching and a low intensity laser for image recording. With the LAS AF application wizard you can choose between different ways to carry out a FRAP experiment. https://www.leica-microsystems.com/science-lab/step-by-step-guide-for-frap-experiments/ Fri, 17 May 2013 13:50:00 +0000 Dr. Jan Schröder https://www.leica-microsystems.com/6600 Confocal Microscopy Live-Cell Imaging Quantitative Imaging Label-free FLIM Many biological samples exhibit autofluorescence. Its often broad spectra can interfere with fluorescent labeling strategies. This application letter demonstrates how autofluorescence can serve as an intrinsic contrast in fluorescence lifetime imaging microscopy (FLIM) resulting in multi-color image stacks. https://www.leica-microsystems.com/science-lab/label-free-flim/ Thu, 09 Aug 2012 22:00:00 +0000 Dr. Constantin Kappel https://www.leica-microsystems.com/6602 Confocal Microscopy Live-Cell Imaging Quantitative Imaging FRET with FLIM FLIM combines lifetime measurements with imaging: lifetimes obtained for each image pixel are color-coded to produce additional image contrast. Thus, FLIM delivers information about the spatial distribution of a fluorescent molecule together with information about its biochemical status or nano-environment. A typical application of FLIM is FLIM-FRET. FRET is a well-established technique to study molecular interactions. It scrutinizes protein binding and estimates intermolecular distances on an Angström scale as well. https://www.leica-microsystems.com/science-lab/fret-with-flim/ Tue, 07 Aug 2012 22:00:00 +0000 Dr. Constantin Kappel https://www.leica-microsystems.com/6609 Confocal Microscopy Quantitative Imaging 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 Imaging 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-1/ Wed, 18 Apr 2012 22:00:00 +0000 https://www.leica-microsystems.com/5082 Confocal Microscopy Quantitative Imaging 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 https://www.leica-microsystems.com/4811 Confocal Microscopy Quantitative Imaging Fluorescence Recovery after Photobleaching (FRAP) and its Offspring FRAP (Fluorescence recovery after photobleaching) can be used to study cellular protein dynamics: For visualization the protein of interest is fused to a fluorescent protein or a fluorescent dye. A region of interest (ROI) can be monitored applying a high amount of light to bleach the fluorescence within the ROI. The following illumination with low light conditions provides insight into the redistribution of molecules via recovery of fluorescence. https://www.leica-microsystems.com/science-lab/fluorescence-recovery-after-photobleaching-frap-and-its-offspring/ Wed, 23 Nov 2011 07:19:37 +0000 Dr. Jan Schröder https://www.leica-microsystems.com/4356 Confocal Microscopy Quantitative Imaging Förster Resonance Energy Transfer (FRET) The Förster Resonance Energy Transfer (FRET) phenomenon offers techniques that allow studies of interactions in dimensions below the optical resolution limit. FRET describes the transfer of the energy from an excited state of a donor molecule to an acceptor molecule. Unlike absorption or emission of photons, FRET is a non-radiative energy exchange and consequently not a variation of light-matter interactions. https://www.leica-microsystems.com/science-lab/foerster-resonance-energy-transfer-fret/ Wed, 09 Nov 2011 09:45:00 +0000 Gabriele Burger