LIFE 212 Lecture Notes - Dark Field Microscopy, Crystallography, Numerical Aperture
30 Mar 2014
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Special lenses (annular stop turret in our lab exercises) Background light is separated from the light diffraction pattern generated by the specimen. Opaque disk inserted between light source and condenser. The length of each optical path differs, which gives the appearance of a 3d relief. Mirrors + excitation + emission filters allow only emitted light of a specific. Multiple images across the vertical z axis are captured and reassembled in 3d. Reduces background fluorescence from unfocused parts of the specimen. Uses high energy electron beam to bombard the specimen (~100 nm thick) with electrons. Dense parts of the specimen do not allow electrons to pass through the specimen. Resolving power = 3 to 10 angstroms ( ) Bombards the surface with electrons to reveal a 3d image. 1 angstrom = 0. 1 nm, 10 = 1 nm. Using a minute probe rather than a high energy xray beam to scan across a.
