Fluorescent Protein

Webinar: Fluorescent Probes and Digital Imaging: Where We Are and Where We're Going



The discovery of green fluorescent protein a half century ago heralded a new and explosive era in microscopy, forever changing the landscape for biology imaging. The ability to fuse a genetically encoded fluorescent probe to an almost-unlimited variety of proteins has enabled scientists to investigate signaling pathways and the movement of intracellular proteins in living cells with unprecedented detail, particularly when coupled with powerful widefield fluorescence and confocal microscopy techniques.



Live-Cell Imaging

Jun 12, 2013

Webinar

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Sample Preparation for GSDIM



The widefield super-resolution technique GSDIM (Ground State Depletion followed by individual molecule return) is a localization microscopy technique that is capable of resolving details as small as 20 nanometers. GSDIM is suitable for a wide range of samples.



Super-Resolution

Jul 27, 2012

Tutorial

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Highlighter Proteins and Fluorescent Timers



The term "Highlighter Proteins" combines a group of genetically altered fluorescent proteins with very special attributes: They can be activated from a non fluorescent state, they can change their emission spectrum or they are even able to be switched "on and off" for a lot of times. This behavior is due to some distinct photo physical attributes. At the single molecule level many fluorescent proteins show an intrinsic blinking activity which can be influenced by external illumination.



Fluorescence Microscopy

Jun 20, 2012

Article

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Fluorescent Proteins – Introduction and Photo Spectral Characteristics



The prospects of fluorescence microscopy changed dramatically with the discovery of fluorescent proteins in the 1950s. The starting point was the detection of the jellyfish Aequorea victoria green fluorescent protein (GFP) by Osamo Shimomura. Hundreds of GFP mutants later, the range of fluorescent proteins reaches from the blue to the red spectrum.



Fluorescence Microscopy

Apr 10, 2012

Tutorial

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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

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FLIM-FRET in Solutions



FRET efficiency can be measured based on fluorescence lifetime microscopy (FLIM). FLIM-FRET allows analysis of molecular interactions both in vitro and in vivo. This article describes the use of FLIM in the time domain (TCSPC) to measure FRET in vitro in a biochemical assay using a Cerulean-Citrine construct.



Confocal Microscopy

Nov 08, 2011

Article

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Cell Cultures and Laser Microdissection



Many of the discoveries that are now being made in cell division and differentiation, the relationships between single cells and cell organelles, treatments of cells with pharmaceutic substances, etc. would not be possible without live cell cultures. Allowing morphological and biochemical observations of single cells under different experimental conditions, they provide a unique source of information.



Laser Microdissection

Oct 07, 2011

Tutorial

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An Introduction in Fluorescence



Fluorescence is widely used in microscopy and an important tool for observing the distribution of specific molecules. Most molecules in cells do not fluoresce. They therefore have to be marked with fluorescing molecules called fluorochromes.



Fluorescence Microscopy

May 31, 2011

Tutorial

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Choose Your Excitation Wavelength



Although time correlated single photon counting (TCSPC) is the method of choice for fluorescence lifetime quantification, it requires dedicated instrumentation including a pulsed laser source, a photon counting card, and a fast detector.



Confocal Microscopy

Apr 13, 2011

Article

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Deep Tissue Imaging



To gain new insight into the fundamental control of cell response to physical changes and to study the dynamics and roles of biological flow during the development of the zebrafish, Dr. Julien Vermot established his lab last year at the Institute of Genetics and Molecular and Cellular Biology (IGBMC) in Strasbourg, France.



Multiphoton Microscopy

Nov 02, 2010

Article

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What is an OPO?



Because light scattering is dependent on the wavelength, better tissue penetration can be achieved by using longer excitation wavelengths. This is where excitation with infrared light, two-photon processes, and the OPO (optical parameter oscillator) can dramatically improve image quality.



Confocal Microscopy

Nov 02, 2010

Article

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The First Supercontinuum Confocal that Adapts to the Sample



Until now, biological and medical research fluorescence imaging in multi-user facilities or institutes has been limited by the type or number of dyes that could be excited. The Leica TCS SP5 X supercontinuum confocal unites the broadband capabilities of the Leica TCS SP5 AOBS® and the freedom and flexibility to select any excitation line within the continuous range of 470 to 670 nm.



White Confocal

Aug 22, 2008

Article

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4D Photoactivation of pa-GFP in Living Cells Using Two-Photon Excitation Laser Scanning Microscopy



We report about two-photon activation of a photoactivatable derivative of the Aequorea Victoria green fluorescent protein (pa-GFP). This special form of the molecule increases its fluorescence intensity when excited by 488 nm after irradiation with high intensity light at 413 nm. Two-photon photoactivation produces an effective real three-dimensional (3D) localization of the molecular switching of pa-GFP in the bright state.



Confocal Microscopy

Oct 15, 2006

Article

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