Biology 2382B Study Guide - Midterm Guide: Hoechst Stain, G1 Phase, Differential Centrifugation
1 Mar 2016
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Section 3: isolation and analysis of cell organelles and molecules. Labeling live cells with fluorescent antibodies or stains. Antibodies are made against specific surface proteins can be linked to fluorophores. Membrane permeable fluorescent dyes can be used to label intracellular structures (i. e. hoechst) stain binds dna in nucleus) Cells with bound antibodies or that have taken up the dyes can now be sorted and counted. T-cells in blood have cd3 and thyl 1 on the cell surface. Create two antibodies, one against each, and tag with different color flurophores. Note: flurophore-linked secondary antibodies can be used in combination with unlabeled primary antibodies. Cells pass single file through a laser beam. Both fluorescent light emitted and scattered are measured by detectors. Individual cells are forced through a nozzle and given a charge proportional to the degree of fluorescence detected. Cells with different electric charges are separated by an. Cells with different electric charges are separated by an electrical field and collected.
