Biology 2382B Lecture Notes - Lecture 3: Hoechst Stain, Western Blot, Fluorescence
28 Feb 2019
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Lecture 3 isolation and analysis of cell organelles and molecules. Important in separating and quantifying cell types in a mixed population: centrifugation for isolation of intracellular organelles. Electrophoresis and western blotting for analysis of proteins. Hoechst stain binds to dna in the nucleus. Sort fluorescence labelled cells using facs: process, cells pass single file through a laser light beam, capillary tube is so small that only on cell at a time can flow through. Laser beam used to excite cells in the tube: cell fluoresces, both fluorescent light emitted and scattered are measured by detectors light emitted is detected and quantified by a fluorescent light detector. Scatterplot from facs: green fluorescence v. red fluorescence, each dot is a cell, by plotting these cells we can determine how many are positive for x or y. How to study organelles & their proteins: need to isolate specific organelles.
