MCB 2210 Lecture Notes - Lecture 3: Immunocytochemistry, Polyclonal Antibodies, Immunofluorescence
6 Sep 2016
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Dramatically improves contrast and allows specific structures to be labeled. Fluorescence when a molecule absorbs light of one wavelength and then re-emits it at a longer wavelength. Fluorophores (fluors) can be linked to various chemicals or biological molecules so as to label specific structures. Because specific molecules glow brightly against a dark background, the contrast of epifluorescence microscopes is excellent. Properties of the lenses used (specifically the numerical aperture, (n. a. the width of the light-cone the objective gathers). Immune proteins that bind to specific proteins: to make an antibody, you obtain pure protein of interest and inject it into an animal, the animal recognizes it as foreign and mounts an immune response o. Individual b-cells will each produce a unique antibody that recognizes a specific: ~ 8-12 amino acid sequence (the epitope). Monoclonal antibodies: made by isolating and cloning a single antibody producing cell, and thus recognize a single epitope.
