ANAT 262 Lecture Notes - Lecture 16: Eyepiece, Immunofluorescence, Tissue Culture
30 Aug 2016
School
Department
Course
Professor
Document Summary
Gfp & immunofluorescence used to localize individual protein. Gfp: cell makes protein for you, can see live. Confocal: camera image different from what is seen in eyepiece. Digital wide-field & confocal are both fluorescent microscopy techniques. If image w/2-3 colours: think of 3 different images visualizing diff proteins. Not all mt in focus, thin cell w/highest resolution objective. Controlled laser focusing to only one point of the specimen at a time. Detector behind adjustable pinhole, only image from in-focus point goes through & detected, out of focus area not visible. Mirrors reflect laser to control point of focus. Computer assembles image, can"t even look through eyepiece. Conventional: thick section most out of focus. Confocal: not physically sectioned, but only see one in-focus section of sample. See sections w/o having to cut, can take images at different sections computer builds 3d image. Fluorescence: can bleach dye if too much light destroy fluorescent molecules.
