ANAT 262 Lecture Notes - Lecture 13: Colocalization, Electron Gun, Inner Membrane
30 Aug 2016
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Cannot really see if two proteins are colocalized, beyond resolution. Partial co-localization: if close together within 200nm, colours will merge. Objective lens above specimen, focus & magnify image. Larger objective: closer to specimen & strength of objective (strength of magnification) At limit of resolution or smaller: look like one dot. Lamda: wavelength of light n: refractive index of medium (maximum 1) Theta: half cone of light from collected by objective (larger lens larger) Lamda: smallest possible n & theta: largest possible (1) Objective lens: magnify & focus image onto projector. Close to nucleus w/electron cloud: interact w/nucleus deflected shadow. If no electrons pass through no image. Biological samples made of light elements, not enough to deflect. Difficult to cut thin layers of soft tissue. Fixation chemicals are cross-linkers: maintain in rigid state. Resin (liquid plastic) hardened in heat or uv. Only get very thin section of cell activity in one image, 200 sections to see entire cell.
