A merged projection of a triple labeled rat pancreatic islet of Langerhans formed form a series of optical sections acquired with the Bio-Rad MRC-600 laser scanning confocal microscope using a krypton/argon ion laser. The three color image was formed by merging individual projections of insulin (green), somatostatin (red), and glucagon (blue) staining. The staining was visualized using FITC-conjugated Donkey anti-Guinea Pig IgG (H+L), 706-095-148; Cy3-conjugated Donkey anti-Rabbit IgG (H+L), 711-165-152; and Cy5-conjugated Donkey anti-Mouse IgG (H+L), 715-175-150. Each of these antibodies was adsorbed against solid-phase immobilized IgGs and serum protiens of other species to minimize cross reactivities. The individual projections were contructed from a series of 25 optical sections acquired at 1.0µm intervals across the top of the islet.
From T.C. Brelje, M.W. Wessendorf, and R.L. Sorenson: Multi-color laser scanning confocal immunofluorescence microscopy: Practical application and limitations. In "Cell Biological Applications of Confocal Microscopy" (B. Matsumoto, ed.), a volume of Methods in Cell Biology 1993. Academic Press, Inc., Orlando, FL.