ANAT 262 Lecture Notes - Lecture 5: Fluorescence Microscope, Bright-Field Microscopy, Confocal Microscopy
Document Summary
Most uses of light microscopy by biologists are to follow particular proteins in cells. Light source emits low wavelength length (high energy) Can be fluorescent dyes in sample to produce longer wavelengths (low energy) Could be weakness, may need normal transmitted light image. Tissue culture cell easy to use for microscopy. If thick piece of tissue section or use confocal microscope. Thin enough to get all cells of tissue in focus. Nucleic acids: use blue dye that integrates into double helix, very simple. Antibodies are y-shaped, tips have two identical variable regions, recognize antigen (recognizes. Antigens can be proteins/short aa sequence, antibodies can be made against any antigen. Fluorescently labeled antibody will give location of antigen. Indirect immunofluorescence: antibody against protein of interest, unlabelled; add fluorescent anti-antibody (e. g. anti mouse antibody) Antigens in cell, antibodies can"t cross pm of cell. Putting hole in pm would make contents leak out.