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Immunofluorescence Microscopy (PDF)
Immunofluorescence microscopy is used to localize specific constituents in tissue sections or immobilized cells using fluorescent tags as labels. The antibody/antigen complex is labeled with any of a variety of fluorochromes emitting light from the near UV to the near IR. Usually FITC, a fluorescein derivative, is used for routine work and shows a yellow/green color. Red is often visualized using Texas Red, rhodamine or Cy3. Blue occurs when AMCA or Cascade Blue is used. Fluorescent microscopes contain limit filters that are optimized for each fluorochrome. Confirm that your microscope is optimized for each dye you plan to use in your experiment. Black and white or color photography or enhanced imaging are often used to record data.
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