What Can You Learn With a Light MicroscopeThis lecture provides an overview of the historical contributions that light microscopy has made to the life sciences and an overview of current exciting developments, such as GFP, super-resolution microscopy, using microscopy to measure biochemical reactions, and imaging of the nervous system
Confocal microscopy is a powerful technique for acquiring three-dimensional images of biological samples. Here I discuss the basic principles of confocal microscopy, with specific discussions of the operation of laser scanning and spinning disk confocal microscopes and of their application to biology.
La Green Fluorescent Protein
GFP <iframe width="640" height="480" src="http://www.youtube.com/embed/Sl2PRHGpYuU" frameborder="0"></iframe>
Zhuang begins her lecture by explaining that the resolution of traditional light microscopy is about 200 nm due to the diffraction of light. This diffraction limit has long hampered the ability of scientists to visualize individual proteins and sub-cellular structures. The recent development of sub-diffraction limit, or super resolution, microscopy techniques, such as STORM, allows scientists to obtain beautiful images of individual labeled proteins in live cells. In Part 2 of her talk, Zhuang gives two examples of how her lab has used STORM; first to study the chromosome organization of E. coli and second, to determine the molecular architecture of a synapse.
FRET: Fluorescence Resonance Energy Transfer
This animation shows how FRET occurs, based on cells transfected with plasmids encoding fluorescent tagged proteins. CFP and YFP were used as donor and acceptor respectively.
Il cervello trasparente
I ricercatori sono riusciti a rendere un intero organo, in questo caso il cervello di un topo, trasparente e permeabile conservandone la struttura in 3D. Utilizzando tecniche di imaging come l’immunofluorescenza, gli scienziati sono inoltre riusciti a visualizzare l’anatomia del cervello e ottenere informazioni preziose sul suo funzionamento.