Biology 2382B Lecture Notes - Lecture 2: Differential Interference Contrast Microscopy, Antonie Van Leeuwenhoek, Fluorescence Microscope
28 Feb 2019
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Document Summary
Fluorescence microscopy: bright-field microscopy and resolution, phase-contrast microscopy, differential interference contrast microscopy, gfp and fluorescent fusion proteins, confocal microscopy, deconvolution. Anton van leeuwenhoek (1632-1723): built many simple, single lens microscopes, focused on the composition of a single drop of some sort of fluid. Features of a modern compound microscope: bright-field microscopy. Limited by a number of factors: 100x by objective lens, 100x by ocular lens, total is a maximum of 1000x, only structures with a high refractive index (ability to bend or slow light) are observable. This is what makes cells observable: but doesn"t provide a lot of detail. Resolution of microscopes: resolution: the ability to distinguish between two very closely positioned objects as separate entities. Resolution = d: distance resolved between 2 points. Nsin : want resolution to be a small number. Oil: : 1/2 angle of light entering objective. The limit of resolution is 0. 2 mm = 200 nm. Wavelength spectrum used in microscopy: visible light: 400-700 nm.
