Lecture2.notes - ii Confocal Microscopy sharp focus with...

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Lecture 2 1/21/10 Reading: Alberts 5th ed: Chapter 8 pg. 510-539.Chapter 9 pgs. 579-593;604- 608 1. Proteins a. sized using SDS-Page b. Proteases can generate fragments c. Western blots identify proteins 2. Looking at cells in the Microscope a. what is resolution? (see pg. 552) b. what is numerical aperture? (see pg. 552) c. What is the limit of resolution? 3. Light Microscope can resolve details 0.2 microM apart. a. resolution: 0.2 microM b. Phase contrast or DIC c. Enhancement and analysis by Digital Techniques d. Fluorescence Microscopy: localize specific molecules i. Immunofluorescence Microscopy, uses antibodies
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Unformatted text preview: ii. Confocal Microscopy: sharp focus with optical sections 4. Electron Microscopy a. resolution: in practical terms, .1nm (1A) b. Immunoglold can localize specific molecules c. Scanning microscopy: a 3d image of the surface of a speciman d. Multiple images can be combined to increase resolution 5. Visualizing Molecules in Living Cells a. uses light microscopy b. Strategies to introduce Membrane-impermeant molecules into cells c. “caged” Molecules d. Fluorescent Proteins can be used to Tag Individual Proteins e. Light Can be Used to Manipulate microscopic Objects...
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