Leica Science Lab - Tag : Diffraction Limit https://www.leica-microsystems.com//science-lab/tag/tags/diffraction-limit/show/Tag/ Article tagged with Diffraction Limit en-US https://www.leica-microsystems.com/15098 Basics in Microscopy Video Talk by Kurt Thorn: The Abbe Diffraction Experiment This lecture describes the famous experiments of Ernst Abbe which showed how diffraction of light by a specimen (and interference with the illuminating light) gives rise to an image and how collection of diffracted light defines the resolution of the microscope. These concepts are demonstrated by using a diffraction grating as a specimen and visualizing and comparing the diffraction pattern in the back focal plane as well as the image in the image plane. https://www.leica-microsystems.com//science-lab/video-talk-by-kurt-thorn-the-abbe-diffraction-experiment/ Mon, 13 Feb 2017 12:12:00 +0000 PhD Kurt Thorn 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/ Mon, 19 Dec 2016 09:29:00 +0000 https://www.leica-microsystems.com/18991 Basics in Microscopy Microscope Resolution: Concepts, Factors and Calculation In microscopy, the term ‘resolution’ is used to describe the ability of a microscope to distinguish detail. In other words, this is the minimum distance at which two distinct points of a specimen can still be seen - either by the observer or the microscope camera - as separate entities. The resolution of a microscope is intrinsically linked to the numerical aperture (NA) of the optical components as well as the wavelength of light which is used to examine a specimen. In addition, we have to consider the limit of diffraction which was first described in 1873 by Ernst Abbe. This article covers some of the history behind these concepts as well as explaining each using relatively simple terminology. https://www.leica-microsystems.com//science-lab/microscope-resolution-concepts-factors-and-calculation/ Fri, 02 Dec 2016 14:09:00 +0000 PhD Martin Wilson https://www.leica-microsystems.com/18088 Super-Resolution Translation Microscopy (TRAM) for Super-Resolution Imaging Super-resolution microscopy is transforming our understanding of biology but accessibility is limited by its technical complexity, high costs and the requirement for bespoke sample preparation. We present a novel, simple and multi-color super-resolution microscopy technique, called translation microscopy (TRAM), in which a super-resolution image is restored from multiple diffraction-limited resolution observations using a conventional microscope whilst translating the sample in the image plane. https://www.leica-microsystems.com//science-lab/translation-microscopy-tram-for-super-resolution-imaging/ Fri, 22 Jul 2016 17:51:00 +0000 https://www.leica-microsystems.com/17673 Confocal Microscopy Super-Resolution Image Restoration and Deconvolution HyVolution – Super-Resolution Imaging with a Confocal Microscope Since the invention of the microscope, there has been continual discussion about the possibility of showing more detailed features of specimens as compared to just magnifying them. In this article we describe the HyVolution concept and how the combination of confocal multiparameter fluorescence imaging at the confocal super-resolution regime with psf-based real deconvolution allows high-speed multicolor imaging with a resolution down to 140 nm. https://www.leica-microsystems.com//science-lab/hyvolution-super-resolution-imaging-with-a-confocal-microscope/ Fri, 01 Apr 2016 07:19:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Constantin Kappel https://www.leica-microsystems.com/16796 Super-Resolution Video: Fluorescence is a State of Mind How to break a fundamental law of physics and win a Nobel Prize to boot. Stefan Hell explains super-resolved fluorescence microscopy for which he shared the 2014 Nobel Prize in chemistry. https://www.leica-microsystems.com//science-lab/video-fluorescence-is-a-state-of-mind/ Thu, 07 Jan 2016 20:45:00 +0000 Prof. Dr. Dr. h.c. Stefan Hell https://www.leica-microsystems.com/16124 Basics in Microscopy A Brief History of Light Microscopy – From the Medieval Reading Stone to Super-Resolution The history of microscopy begins in the Middle Ages. As far back as the 11th century, plano-convex lenses made of polished beryl were used in the Arab world as reading stones to magnify manuscripts. However, the further development of these lenses into the first microscopes cannot be attributed to any one person. It took the ideas and designs of many scientists and scholars to produce instruments capable of strong magnification. https://www.leica-microsystems.com//science-lab/a-brief-history-of-light-microscopy-from-the-medieval-reading-stone-to-super-resolution/ Tue, 08 Sep 2015 17:41:00 +0000 Wymke Ockenga 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/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/15237 Super-Resolution Live-Cell Imaging Pathways to Optical STED Microscopy STED nanoscopy has evolved to a highly versatile tool for the observation of the living cell, more and more finding its way into state-of-the-art optical imaging facilities in biomedical research institutes. https://www.leica-microsystems.com//science-lab/pathways-to-optical-sted-microscopy/ Thu, 12 Mar 2015 15:49:00 +0000 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/12945 Super-Resolution GSDIM Publication List GSDIM microscopy is a widefield super-resolution technique based on the localization of fluorophores with nanometer precision. With its help a lateral resolution of down to 20 nm can be achieved, whereas the new 3D feature even shrinks axial resolution to 50 nm. Here we provide a collection of publications around that super-resolution microscopy method also called dSTORM. https://www.leica-microsystems.com//science-lab/gsdim-publication-list/ Sun, 15 Feb 2015 10:25:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/15079 Super-Resolution Confocal Microscopy Image Restoration and Deconvolution Super-Resolution – On a Heuristic Point of View About the Resolution of a Light Microscope Since super-resolution has become one of the most favored methods in biomedical research, the term has become increasingly popular. Still, there is much of confusion about what is super-resolution and what is resolution at all. Here, the classical view of microscopic resolution is discussed and some techniques that resolve better than classical are briefly introduced. The picture on the right shows the intensity distribution of an image of two points whose distance is just the Rayleigh criterion (false color coding). https://www.leica-microsystems.com//science-lab/super-resolution-on-a-heuristic-point-of-view-about-the-resolution-of-a-light-microscope/ Wed, 21 Jan 2015 16:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/15059 Super-Resolution Finding, Defining and Breaking the Diffraction Barrier in Microscopy – A Historical Perspective Diffraction plays a crucial role in microscopy as it prevents the recording of arbitrarily sharp images with conventional light microscopes. Many names are connected with the notion of diffraction and the definition of resolution. An overview over the contributions of the different scientists to the recognition and definition of the diffraction barrier in the past centuries is given and the recent developments that led to breaking this barrier are portrayed. https://www.leica-microsystems.com//science-lab/finding-defining-and-breaking-the-diffraction-barrier-in-microscopy-a-historical-perspective/ Mon, 12 Jan 2015 15:42:00 +0000 https://www.leica-microsystems.com/14451 Super-Resolution Super-Resolution Microscopy – Get Your Free e-Book for Download Until recently, the diffraction of light had placed a fundamental limit on how far biologists could peer into cells with optical microscopes, preventing them from resolving features less than 250 nm in size, missing critical structures within cells. Over the past 20 years scientists have developed several ingenious techniques allowing them to resolve features as small as 20 nm. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-get-your-free-e-book-for-download/ Wed, 19 Nov 2014 15:10:00 +0000 https://www.leica-microsystems.com/14647 Super-Resolution 3D STED (Stimulated Emission Depletion) Microscopy The resolution needed to image subcellular architecture and dynamics in light microscopy is hindered by the diffraction limits as described by Ernst Abbe. Simply stated, structures smaller than 200 nanometers are lost in a blur. However, the field of super-resolution microscopy has produced methods to obtain resolution beyond this limit. Leica Microsystems has pioneered this field and offers the Leica TCS SP8 STED 3X for 3D Stimulated Emission Depletion microscopy. STED instantly produces super-resolution images, compatible with the dynamics of living cells, without the need for post-processing. https://www.leica-microsystems.com//science-lab/3d-sted-stimulated-emission-depletion-microscopy/ Tue, 14 Oct 2014 14:03:00 +0000 PhD Christopher Vega https://www.leica-microsystems.com/14623 Super-Resolution Nobel Prize in Chemistry for Achievements in Super-Resolution Microscopy On October 8th 2014, The Royal Swedish Academy of Sciences has decided to award the Nobel Prize in Chemistry for 2014 to Eric Betzig, Stefan W. Hell and William E. Moerner "for the development of super-resolved fluorescence microscopy". For a long time optical microscopy was held back by a presumed limitation: that it would never obtain a better resolution than half the wavelength of light. Helped by fluorescent molecules the Nobel Laureates in Chemistry 2014 ingeniously circumvented this limitation. Their ground-breaking work has brought optical microscopy into the nanodimension. https://www.leica-microsystems.com//science-lab/nobel-prize-in-chemistry-for-achievements-in-super-resolution-microscopy/ Thu, 09 Oct 2014 13:25:00 +0000 Nobel Media AB 2014 https://www.leica-microsystems.com/13795 Super-Resolution Video Interview with Stefan Hell, the Inventor of Super-Resolution Professor Stefan Hell is director at the Max Planck Institute for Biophysical Chemistry and head of the department of NanoBiophotonics in Goettingen and widely considered as the father of super-resolution. His inventions of 4Pi and STED microscopy were turned into the first commercial super-resolution microscopes available by Leica Microsystems in 2004 and 2007. https://www.leica-microsystems.com//science-lab/video-interview-with-stefan-hell-the-inventor-of-super-resolution/ Wed, 08 Oct 2014 18:14:00 +0000 Prof. Dr. Dr. h.c. Stefan Hell, Dipl. oec.-troph. Anja Schué, PhD Isabelle Köster https://www.leica-microsystems.com/11763 Basics in Microscopy Video Talk by Jeff Lichtman: Resolution in Microscopy – Wave Optics and the Diffraction Limit Light has properties of particles and waves. Understanding the wave nature of light is essential to understanding the workings of a microscope. This lecture describes Huygens Wavelets, constructive/destructive interference, and diffraction. https://www.leica-microsystems.com//science-lab/video-talk-by-jeff-lichtman-resolution-in-microscopy-wave-optics-and-the-diffraction-limit/ Thu, 14 Aug 2014 12:48:00 +0000 Dr. Jeff Lichtman https://www.leica-microsystems.com/13950 Super-Resolution Neuroscience STED Microscopy of Living Cells – New Frontiers in Membrane and Neurobiology Recent developments in fluorescence far-field microscopy such as STED microscopy have accomplished observation of the living cell with a spatial resolution far below the diffraction limit. Here, we briefly review the current approaches to super-resolution optical microscopy and present the implementation of STED microscopy for novel insights into live cell mechanisms, with a focus on neurobiology and plasma membrane dynamics. https://www.leica-microsystems.com//science-lab/sted-microscopy-of-living-cells-new-frontiers-in-membrane-and-neurobiology/ Tue, 12 Aug 2014 12:11:00 +0000 Prof. Christian Eggeling https://www.leica-microsystems.com/13527 Super-Resolution Video Talk on Super-Resolution: Overview and Stimulated Emission Depletion (STED) Microscopy Historically, light microscopy has been limited in its ability to resolve closely spaced objects, with the best microscopes only able to resolve objects separated by 200 nm or more. This limit is known as the diffraction limit. In the last twenty years, a number of techniques have been developed that allow resolution beyond the diffraction limit. https://www.leica-microsystems.com//science-lab/video-talk-on-super-resolution-overview-and-stimulated-emission-depletion-sted-microscopy/ Thu, 05 Jun 2014 17:05:00 +0000 Prof. Dr. Dr. h.c. Stefan Hell https://www.leica-microsystems.com/11024 Super-Resolution Interview with Stefan Hell: Breaking Nanoscale Boundaries Stefan Hell is credited with having conceived and applied the first viable concept for breaking Abbe’s diffraction limit in a light-focusing microscope. He is a scientific member of the Max Planck Society and a director at the Max Planck Institute for Biophysical Chemistry in Göttingen, Germany, where he currently leads the department of nanobiophotonics. https://www.leica-microsystems.com//science-lab/interview-with-stefan-hell-breaking-nanoscale-boundaries/ Wed, 09 Oct 2013 13:29:00 +0000 Dr. Peter Andersen https://www.leica-microsystems.com/9043 Super-Resolution STED and GSDIM: Diffraction Unlimited Resolution for all Types of Fluorescence Imaging This article gives an overview of two different types of superresolution techniques. Stimulated emission depletion (STED) microscopy is a versatile and fast method that is based on point scanning microscopy – usually an extension of a confocal microscope. Ground state depletion imaging (GSDIM) is a parallel recording widefield approach that explores inherent switching of fluorochromes and typically comes with a TIRF microscope. The two methods use very different approaches to reach the same goal: to see more details in light microscopes than possible when diffraction limited. https://www.leica-microsystems.com//science-lab/sted-and-gsdim-diffraction-unlimited-resolution-for-all-types-of-fluorescence-imaging/ Thu, 28 Feb 2013 23:00:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/6497 Widefield Microscopy Super-Resolution Widefield Super-Resolution with GSDIM Great advancements in biology have been possible by using fluorescence microscopy. So far, the resolution of the images was limited due to physical constraints. In the past couple of years, new methods evolved circumventing these limitations and bringing fluorescence microscopy to a new level of resolution, boosting the possibilities in science with fluorescence microscopes. https://www.leica-microsystems.com//science-lab/widefield-super-resolution-with-gsdim/ Wed, 11 Jul 2012 22:00:00 +0000 Dr. Marko Lampe https://www.leica-microsystems.com/5815 Super-Resolution Gated STED Microscopy with CW-STED Lasers Among the major steps in the development of Stimulated Emission Depletion (STED) microscopy, the demonstration of the use continuous wave lasers (CW-STED) was certainly contributing the most to a wide dissemination of the method due to the affordability and elegant simplicity of this implementation. Nevertheless, CW-STED was so far not reaching the same spatial resolution as pulsed-lasers STED configurations. A recent investigation on the time-course of the fluorescence emission probability in CW-STED has revealed the benefit of using a gated fluorescence detection (gSTED) to improve further the resolution of C-S https://www.leica-microsystems.com//science-lab/gated-sted-microscopy-with-cw-sted-lasers/ Sun, 01 Apr 2012 22:00:00 +0000 Ph.D. Giuseppe Vicidomini, Gael Moneron 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/5331 Neuroscience Super-Resolution Sharp Live Images from the Mouse Brain To explore the most intricate structures of the brain in order to decipher how it functions – Stefan Hell’s team of researchers at the Max Planck Institute for Biophysical Chemistry in Göttingen has made a significant step closer to this goal. Using the STED microscopy developed by Hell, the scientists have, for the first time, managed to record detailed live images inside the brain of a living mouse. https://www.leica-microsystems.com//science-lab/sharp-live-images-from-the-mouse-brain/ Sun, 04 Mar 2012 23:00:00 +0000 Dr. Sebastian Berning, Dr. Katrin Willig, Dr. Heinz Steffens, Dr. Payam Dibaj, Prof. Dr. Dr. h.c. Stefan Hell https://www.leica-microsystems.com/4820 Super-Resolution Videos on STED Microscopy and its Inventor Stefan Hell Here we present an collection of videos telling the story of the invention of nanometer-scale microscopy by Stefan Hell and explaining the basic principle of STED microscopy. https://www.leica-microsystems.com//science-lab/videos-on-sted-microscopy-and-its-inventor-stefan-hell/ Thu, 08 Dec 2011 14:33:00 +0000 https://www.leica-microsystems.com/4621 Super-Resolution Widefield Microscopy How Widefield Super-Resolution GSDIM Images are Created The localization microscopy technique GSDIM is a proven technology to achieve super-resolution images with a resolution of up to 20 nm. In the following tutorial we will describe the basic principles and features of GSDIM. https://www.leica-microsystems.com//science-lab/how-widefield-super-resolution-gsdim-images-are-created/ Mon, 31 Oct 2011 17:17:00 +0000 Dr. Marko Lampe https://www.leica-microsystems.com/4600 Super-Resolution Step by Step Guide to the Molecular Basics of GSDIM Microscopy Ground state depletion microscopy followed by individual molecule return (GSDIM) is a super-resolution technique based on single molecule localization (Localization Microscopy). To localize single molecules and create a high resolution image the ensemble of overlapping fluorophores (in a diffraction-limited setup) has to be broken up. Individual fluorophores must be temporally "separated" to allow high precision detection of single molecules. This tutorial will explain the molecular basics of GSDIM. https://www.leica-microsystems.com//science-lab/step-by-step-guide-to-the-molecular-basics-of-gsdim-microscopy/ Fri, 28 Oct 2011 16:17:00 +0000 Dr. Marko Lampe https://www.leica-microsystems.com/4438 Super-Resolution Super-Resolution GSDIM Microscopy The nanoscopic technique GSDIM (ground state depletion microscopy followed by individual molecule return) provides a detailed image of the spatial arrangement of proteins and other biomolecules within the cell. There is now a first commercial system (the Leica SR GSD) on the market that is helping to make the GSDIM technique available to a wider group of users in research labs and imaging centers. https://www.leica-microsystems.com//science-lab/super-resolution-gsdim-microscopy/ Thu, 20 Oct 2011 22:00:00 +0000 PhD Tamara Straube, Alexandra Elli, Prof. Ralf Jacob https://www.leica-microsystems.com/3911 Super-Resolution Unlimited Resolution - STED STED uses a differential method of two different diffraction patterns, where one pattern excites and the second pattern de-excites fluorochromes. The residual excited area is controllable by intensity down to (theoretically) zero – unlimited resolution. https://www.leica-microsystems.com//science-lab/unlimited-resolution-sted/ Thu, 30 Jun 2011 22:00:00 +0000 Dr. Rolf T. Borlinghaus 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/14660 Super-Resolution Interview with Stefan Hell – For me, Pioneering is … Our interview partner on the topic of Pioneering is Prf. Stefan Hell, a scientific member of the Max Planck Society and director at the Max Planck Institute for Biophysical Chemistry in Göttingen, Germany, where he heads the Department of Nanobiophotonics. In addition to further chairs and memberships, he is also head of the High resolution Optical Microscopy research group, a partnership department of the German Cancer Research Center (DFKZ). https://www.leica-microsystems.com//science-lab/interview-with-stefan-hell-for-me-pioneering-is/ Sun, 31 Aug 2008 22:00:00 +0000