Lecture 1--microscopy

Lecture 1-microscopy

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Unformatted text preview: luminating the entire sample, the whole image can onli work with fixed be reconstructed from these points s pecimens Huang lab, UCSF How does super-resolution work? --Stochastic optical resolution microscopy (STORM) sharper images microtubules and mitochondria, from Huang lab, UCSF filament techniqu es imptrove left to right all use fluorophores which are 10nm away from where it is s uposed to be since attache by Limit of conventional light microscopy: 200-250 nm Limit of super-resolution microscopy: 10 nm Limit of conventional light microscopy: 200-250 nm Limit of super-resolution microscopy: 10 nm Limit of electron microscopy: 0.05 nm limited by wavlength of electron not light, which is much smaller Transmission Electron Microscopy 2 types of electron microscioy For TEM very thin tissue slices are placed on a grid and stained with heavy metals that deflect electrons Images are formed from electrons transmitted light microscopy needed through the specimen fluorescence to be able to stai and see contrast, electrons easily pass through tissues, things that reflect electrons are heavy metals s o need that so that not all of tissue is the same Transmission Electron Microscopy For TEM very thin tissue slices are placed on a grid and stained with heavy osmium tetroxide metals Images are formed from electrons transmitted through the specimen In electron microscopy, sections are cut with an ultramicrotome Tissue dissected out and placed in the fixing solution microtome Specimen vial specimen oven ultramicrotome made from dimands and can slice even s maller The sections are picked off the surface with a copper grid Sections for Transmission Electron Microscope slice of head of fish Zooming in with Transmission Electron Microscope axon i cross section s urrounded by skin c ells have to be done on thin s pecimens black and white be able to recognixe these Scanning Electron Microscope Electron gun Condenser lens Beam deflector Objective lens Detector Electrons from specimen Images are formed from the electrons that have bounced coat whole sample, with meta (silver, uranium), off the surface of the specimen. focus light on specimen, reflect light off s urfe=ace detected by detector celia of hair cell in iner ear TEM and SEM provide different perspectives into cellular structure flat prop...
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This note was uploaded on 02/23/2014 for the course MCDB 165A taught by Professor Iruela-arispe during the Winter '08 term at UCLA.

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