DernburgLecture2_2-1-10_forprinting

DernburgLecture2_2-1-10_forprinting - Light microscopy has...

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Microtubules Light microscopy has a limited ability to reveal fne detail 1 Point Spread Function (PSF) Problem: A “point source” o± light does not look like a point through the microscope. It is blurred by the “Point Spread Function,” or PSF 2 Every point o± light that makes up a ²uorescent object behaves in the same way, resulting in a blurred image 3 Microtubules Light microscopy has a limited ability to reveal fne detail 4 Microtubules as seen through a microscope Light microscopy has a limited ability to reveal fne detail Resolution limit: the distance between two objects required ±or them to be seen as separate ~200-300 nm 5 **** Permits dynamic and in vivo analysis**** o± biological processes Green Fluorescent Protein (GFP) Comes ±rom a jellyfsh, Aequorea victoria Gene has been cloned and trans±erred into a wide variety o± “heterologous” expression systems … including Drosophila, mammalian cells, C. elegans , yeast, zebrafsh etc. etc. neurons zebrafsh pigs!!!? 6
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2008 Nobel Prize in Chemistry Osamu Shimomura Roger Tsien Marty Chalfe 7 Tyrosine 66 Glycine 65 Serine 67 Both chemical and biochemical fuorophores contain extended networks oF conjugated double bonds G±P ±luorescein Rhodamine The “ β -barrel” structure oF G±P provides a special environment inside the living cell that enables the fuorophore to work 8 Variants oF Green ±luorescent Protein and DsRed have been engineered to have diFFerent excitation and emission spectra, and other useFul properties
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This note was uploaded on 02/16/2010 for the course MCB 74282 taught by Professor Professordernburg during the Spring '10 term at University of California, Berkeley.

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DernburgLecture2_2-1-10_forprinting - Light microscopy has...

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