Biology 2382B Lecture Notes - Lecture 3: Hoechst Stain, Fluorophore, Mortar And Pestle
27 Jun 2017
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Lecture 3: isolation and analysis of cell organelles and molecules: We separate proteins based on: size, net charge, affinity. We add membrane particle which will bind the nuclear dna and ensure it has a nucleus: so now we have 3 types of dyes and can quantify the types of cells labeled. Fluorescent activated cell sorting (facs): cells pass single file through a laser light beam, both fluorescent light. 3. 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 electric field and collected. Cells are labeled in a solution mixture and then loaded into tubes. Tube gets progressively smaller as cells do down this glass tube: laser light can illuminate at specific wavelengths which will cause excitation of the fluorophore, light emitted can be detected by specific detectors.
