Biology 2382B Lecture Notes - Lecture 4: Dichroic Filter, Fluorophore, Primary And Secondary Antibodies
15 Mar 2015
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Need: good eyesight & good grinding lens. Light (passes through specimen) is focused and collected by objective lens (100x) (magnifying) Annular diaphragm creates cone of light (limits light passing through specimen) Only images with high refractive indexes are visible (greater bending of light) Bright with dark areas (slower light = more refraction) All light that passes through specimen passes through objective lens. As it hits phase plate (coating in film) gets shorten another wavelength. Recombine wavelength with the light that is unimpeded gives you contrast. Detail: ability of wavelengths to cancel each other out. Areas of the cell that refract the light better create darker areas. Light is 3d wave can have vertical, horizontal, and in between angles. Filters (polarizers) that allow only vertical light or horizontal light (limit the type of light passing through the specimen) Shows shadow effect (makes membranes etc show up) Halo effect is eliminated (waves are no longer mixed up)
