BIOL 311 Lecture Notes - Lecture 12: Scanning Electron Microscope, Immunochemistry, Ferritin

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Freeze fracture: allows you to view internal surface of membranes, procedure, freeze cells, fracture follows line of least resistance. Will go around proteins: stain with platinum. Ultrastructural immunochemistry: use antibodies to localize proteins (or other molecules) in cells, attach gold particle to antibody, double label technique, add 2 different size gold. Ex: 20 nm and 5 nm: use gold and ferritin. Ferritin fuzzy : example, ldl internalization, pathway. Attach to receptor on cell membrane label ldl with small gold and receptor with large. Once ldl and receptor are in cell, they separate. Ultrastructural autoradiography: use radioactive probes, ex: acetylcholine receptors. Electron tomography: computer reconstract 3rd dimension, serial sections, image and compile each. Laser capture microscopy (dissection: purpose identify cell(s) of interest and dissecting/removing cell for further analysis, nih developed it, steps, tissue section, overlay section with ethylene vinyl acetate film, laser spot of 7. 5-30 m. Melt film on cell: remove cell, analyze cell.

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