STED

STED and GSDIM: Diffraction Unlimited Resolution for all Types of Fluorescence Imaging

$STED and GSDIM: Diffraction Unlimited Resolution for all Types of Fluorescence Imaging$

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.

Super-Resolution

Mar 01, 2013

Abstract

Webinar: From Live Cell Imaging to Super-Resolution

The beauty of cells, magnified and resolved by light microscopy, has been fascinating investigators since the 19th century. Today, functional research in living cells is often a prerequisite for biological studies. And keeping the cells close to the necessary conditions whilst under microscopical observation is key. In this webinar, Dr. Christoph Bauer will share his experiences of live cell imaging and talk about how the microscopy challenges, such as optical aberrations at 37°C or focus drift in long-term experiments, can be addressed.

Super-Resolution

Feb 07, 2013

Webinar

Gates Open for Improved Confocal Fluorescence and Super-Resolution STED

True confocal microscope systems feature single-point illumination and single-point detection. The method is called "optical sectioning" since the generated image contains only information from the focal plane. The serial detection offers highly efficient and low-noise sensors for signal conversion. Although the nonparallel detection is not conducive to high-speed imaging, modern scanning concepts allow frame rates above 400 frames per second at reasonable noise levels. This is by far enough for most applications, including the monitoring of fast ion-transport phenomena in living material.

Super-Resolution

Jan 24, 2013

Tutorial

Mitochondrial DNA Molecules Are Packaged Individually

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At the Max Planck Institute for Biology of Ageing in Cologne founded in 2008, three departments are working on the molecular, physiological and evolutionary mechanisms of the ageing of cells, tissue and organisms. The long-term aim of the research is to help people age more healthily. Here, Dr. Christian Kukat from the department of Professor Nils-Göran Larsson is examining the role played by mitochondria in the ageing process. In close collaboration with Dr. Christian Wurm from the research group of Professor Stefan Jakobs at the Max Planck Institute for Biophysical Chemistry in Göttingen and researchers at the University of Gothenburg, Sweden, they harnessed super-resolution STED microscopy to find out new and surprising facts about the mitochondrial nucleoid.

Super-Resolution

Jan 07, 2013

Article

Quick Guide to STED Sample Preparation

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This document is intended as a quick reference guide for the most common questions regarding the preparation of samples for the Leica SP8 TCS STED, by briefly explaining the theoretical basics required from samples for stimulated emission depletion. It provides information about the most common immuno-fluorescence labeling techniques, working mounting media, basic quality optimization procedures and experiment designs. It also contains a detailed list of reagents, antibodies and disposables frequently and successfully used in super-resolution STED microscopy. In summary, this guide is meant to present the necessary knowledge, including some tips and tricks, to users that are preparing their first super-resolution microscopy sample.

Super-Resolution

Nov 14, 2012

Tutorial

Abstracts of the 2nd European Super-Resolution User-Club Meeting

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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.

Super-Resolution

Nov 02, 2012

Article

Depletion and Emission Beam Paths in STED Microscopy

STED is one technology, and it requires already additional beam-routings as compared to pure fluorescence imaging. The Leica TCS SP5 offers configurations with STED, confocal and multiphoton imaging in the very same instrument. This unique concept unites technologies of super-resolution fluorescence, multichannel confocal fluorescence, multiphoton-excited fluorescence and second-harmonic or higher order nonlinear image generation. The various beam-paths are shown and explained in this tutorial.

Super-Resolution

May 21, 2012

Tutorial

Stimulated Emission Depletion (STED)

Among the new super-resolution technologies, stimulated emission depletion (STED) is the most versatile concept. Whether the sample is tissue, e.g. muscle striation details, classical cytoskeletons, nuclear proteins, yeast or bacterial details: STED serves for understanding structure and function by showing the very finest details.

Super-Resolution

May 21, 2012

Tutorial

Beam Paths of a Continuous Wave STED Microscope

This tutorial gives on overview of the excitation of an detection light path with a continuous wave (CW) STED microscope.

Super-Resolution

May 14, 2012

Tutorial

STED Super-Resolution Microscopy

This tutorial focusses on the principles of STED super-resolution microscopy. The underlying photo physical processes are explained - e.g. with the help of a Jablonski diagram - and the integration into a confocal laser scanning microscope.

Super-Resolution

May 02, 2012

Tutorial

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

Super-Resolution

Apr 02, 2012

Article

Webinar: Beyond the Diffraction Limit

$Webinar: Beyond the Diffraction Limit$

Super-resolution imaging is expanding our understanding of cellular structures and protein dynamics. First suggested more than 20 years ago, fluorescence techniques capable of resolution in the tens of nanometer range are becoming increasingly commonplace today. In this special web seminar, viewers will learn the basic principles behind several of the most commonly used super-resolution imaging techniques (STED, PALM, STORM, and GSD), hear specific examples where super-resolution approaches have provided unique biological insights, and get a glimpse of what the future holds for super-resolution studies.

Super-Resolution

Mar 28, 2012

Webinar

Visualization of the Natural Killer Cell Immune Synapse by Super-Resolution Nanoscopy

Natural killer (NK) cells are innate immune effectors that recognize and kill virally infected and tumorigenic cells. Central to their effector function is the formation of an immunological synapse with a target cell followed by directed secretion of cytolytic granules.

Super-Resolution

Mar 09, 2012

Article

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.

Mar 05, 2012

Abstract

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.

Live-Cell Imaging

Feb 24, 2012

Tutorial

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.

Super-Resolution

Dec 08, 2011

Tutorial

Subcellular Localization of AKT and Tubulin using Super-Resolution Microscopy

Stimulated Emission Depletion microscopy, or STED nanoscopy, is a technique that uses the non-linear de-excitation of fluorescent dyes to overcome the resolution limit imposed by diffraction encountered with standard confocal laser scanning microscopes and conventional far-field optical microscopes[1]. Compared to traditional confocal microscopy, STED offers exceptional improvements in resolution allowing visualization of cellular events at unprecedented levels.

Super-Resolution

Dec 01, 2011

Article

Modern Fluorescent Proteins and their Biological Applications

Here we present two review articles on fluorescent proteins and their biological applications. These first article reviews our current knowledge of blue, green, and red chromophore formation in permanently emitting FPs, photoactivatable FPs, and fluorescent timers. The second article focuses on novel monomeric RFPs and their application for studying gene expression, nuclear localization, and dynamics using advanced imaging.

Fluorescence Microscopy

Nov 18, 2011

Abstract

Webinar: Applications, Labeling Strategies and Fluorophores for Super-Resolution

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The free online webinar on super-resolution presented by Leica Microsystems in association with Microscopy & Analysis took place on Tuesday, 15 November 2011. Register and view the webinar on demand.

Nov 15, 2011

Webinar

Neuroscience and Microscopy

Neurobiology, the science of nerves and the brain, has mainly been driven forward in the last 200 years by microscopic investigations. The structures of cellular and subcellular structures, interaction and the three-dimensional assembly of neurons were made visible by various microscopy techniques. The optical microscope is also a necessary tool for visualizing micropipettes in electrophysiological measurements. Thirdly, many types of functional imaging are performed by means of optical microscopy.

Nov 09, 2011

Article

Abstracts of the First European Super-Resolution User Club Meeting

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The first European Super-resolution User Club meeting took place from October 27 to 29 in Göttingen, Germany. Prof. Stefan Hell, the inventor of the STED technology, has hosted this first meeting. The user club is aimed at pioneering researchers from the European scientific community, who are early adopters and developers of super-resolution techniques.

Super-Resolution

Oct 11, 2011

Abstract

Webinar: Super-Resolution

In 1873, Ernst Abbe developed a theory that defined the limit of resolution of the light microscope. Following suit from astronomy, Abbe defined resolution as the ability to resolve, as separate, two point sources of light. The Abbe limit of 200–300 nm is based upon the ability of the light microscope to collect only a subset of spatial frequencies and the physiological properties of the human eye.

Super-Resolution

Sep 08, 2011

Webinar

Unlimited Resolution

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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.

Super-Resolution

Jul 01, 2011

Tutorial

A STED-y Route to Commercialization

The diffraction barrier responsible for a finite focal spot size and limited resolution in far-field fluorescence microscopy has been fundamentally broken.

Super-Resolution

Jun 16, 2011

Abstract

STED Publication List

Publications using STED microscopy

Super-Resolution

Jun 09, 2011

Article

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