Biology 2382B Lecture Notes - Lecture 3: Centrifugal Force, Differential Centrifugation, Cell-Cycle Analysis
11 Feb 2017
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Lecture 3 isolation and analysis of cell organelles and molecules. Labeling live cells with fluorescent antibodies or stains. Antibody specifically recognizes each of the transmembrane proteins: conjugate antibodies with fluorophores or fluorochromes, recognize proteins only on the outside of cells. Antibodies made against specific cell surface proteins can be linked to fluorophores. Cells with bound antibodies or that have taken up the dyes can now be sorted and counted. 3 potential fluorescent parameters that we can monitor: thy1 expressed in cells will have more green, cd3 will have more red, nucleated cells will be blue. Note: fluorophore-linked secondary antibodies could also be used in combination with unlabeled primary antibodies. 2 strategies: antibodies, cell permeable dyes that cross into cell and bind to dna. Have illumination sources (lasers) that are focused on a tube and the tube transports fluids with cells that we"ve labelled (osmotically balanced)
