Leica Science Lab - Tag : Gated Detection https://www.leica-microsystems.com//science-lab/tag/?tx_leicaacademy_pi4%5Baction%5D=show&tx_leicaacademy_pi4%5Bcontroller%5D=Tag&tx_leicaacademy_pi4%5Btag%5D=677&cHash=0dbb3e3f93c073331009bd404ae86502 Article tagged with Gated Detection en-US https://www.leica-microsystems.com/24510 Confocal Microscopy You Shall Not Pass! Time-gated Detection Takes Care of Endogenous Fluorescence in Plant Research. Yutaka Kodama is Associate Professor at the Biosciences Education Research Center, Utsunomiya University, Japan. He graduated from Saga University, Faculty of Agriculture, in 2004 and received his PhD from the Graduate School of Biological Science at the Nara Institute of Science and Technology in 2007. https://www.leica-microsystems.com/science-lab/you-shall-not-pass-time-gated-detection-takes-care-of-endogenous-fluorescence-in-plant-research/ Tue, 13 Nov 2018 23:00:00 +0000 https://www.leica-microsystems.com/18271 Super-Resolution Quantitative Imaging 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/17611 Super-Resolution Multiphoton Microscopy Live-Cell Imaging 4Pi-RESOLFT Nanoscopy Here we apply the 4Pi scheme to RESOLFT nanoscopy using two-photon absorption for the on-switching of fluorescent proteins. We show that in this combination, the lobes are so low that low-light level, 3D nanoscale imaging of living cells becomes possible. Our method thus offers robust access to densely packed, axially extended cellular regions that have been notoriously difficult to super-resolve. Our approach also entails a fluorescence read-out scheme that translates molecular sensitivity to local off-switching rates into improved signal-to-noise ratio and resolution. https://www.leica-microsystems.com/science-lab/4pi-resolft-nanoscopy/ Fri, 26 Feb 2016 12:41:00 +0000 https://www.leica-microsystems.com/15928 Super-Resolution Gated STED Microscopy with Time-gated Single-photon Avalanche Diode The maximization of the useful (within the time gate) photon flux is then an important aspect to obtain super-resolved STED images. Here we show that by using a fast-gated single-photon avalanche diode (SPAD), i.e. a detector able to rapidly (hundreds picoseconds) switch-on and -off can improve significantly the signal-to-noise ratio (SNR) of the gated STED image. In addition to an enhancement of the image SNR, the use of the fast-gated SPAD reduces also the system complexity. We demonstrate these abilities both on calibration and biological sample. https://www.leica-microsystems.com/science-lab/gated-sted-microscopy-with-time-gated-single-photon-avalanche-diode/ Tue, 25 Aug 2015 17:24:00 +0000 https://www.leica-microsystems.com/16176 Super-Resolution Quantitative Imaging 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/13128 Super-Resolution Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy Here we illustrate the protocol for imaging by two-color STED nanoscopy the cytotoxic immune synapse of NK cells recapitulated on glass. Using this method we obtain sub-100 nm resolution of synapse proteins and the cytoskeleton. https://www.leica-microsystems.com/science-lab/visualization-of-the-immunological-synapse-by-dual-color-time-gated-stimulated-emission-depletion-sted-nanoscopy/ Fri, 28 Mar 2014 16:34:00 +0000 Ph.D. Emily M. Mace, Ph.D., MD Jordan S. Orange https://www.leica-microsystems.com/13103 Super-Resolution A New Filtering Technique for Removing Anti-Stokes Emission Background in Gated CW-STED Microscopy Stimulated emission depletion (STED) microscopy is a prominent approach of super-resolution optical microscopy, which allows cellular imaging with so far unprecedented unlimited spatial resolution. The introduction of time-gated detection in STED microscopy significantly reduces the (instantaneous) intensity required to obtain sub-diffraction spatial resolution. https://www.leica-microsystems.com/science-lab/a-new-filtering-technique-for-removing-anti-stokes-emission-background-in-gated-cw-sted-microscopy/ Tue, 25 Mar 2014 20:34:00 +0000 https://www.leica-microsystems.com/10208 Super-Resolution STED Nanoscopy with Time-Gated Detection: Theoretical and Experimental Aspects In a stimulated emission depletion (STED) microscope the region in which fluorescence markers can emit spontaneously shrinks with continued STED beam action after a singular excitation event. This fact has been recently used to substantially improve the effective spatial resolution in STED nanoscopy using time-gated detection, pulsed excitation and continuous wave (CW) STED beams. https://www.leica-microsystems.com/science-lab/sted-nanoscopy-with-time-gated-detection-theoretical-and-experimental-aspects/ Mon, 08 Jul 2013 08:48:00 +0000 Ph.D. Giuseppe Vicidomini https://www.leica-microsystems.com/10111 Confocal Microscopy Multiple Microscopy Modes in a Single Sweep with Supercontinuum White Light Lasers have been critical to the advancement on confocal microscopy, and the white light laser (WLL) offers particular advantages. Finessing WLL output for bioimaging is a complex task, though, and traditional approaches retain key limitations. But acousto-optical beamsplitting enables smoother operation, leading to enhanced microscopy capabilities. https://www.leica-microsystems.com/science-lab/multiple-microscopy-modes-in-a-single-sweep-with-supercontinuum-white-light/ Fri, 14 Jun 2013 14:09:00 +0000 Dr. Lioba Kuschel, Dr. Rolf T. Borlinghaus