A NOVEL, RAPID FLAT-MOUNTING TECHNIQUE FOR VISUALIZING ANTIBODY LABELING IN THE RETINA

Complete analysis of retinal tissue is difficult because it consists o f a thin neural tissue spread across the back of a hemispheric surface . Conventional sectioning in a plane parallel to a central axis of sym metry produces a large number of samples, each containing only a small amount of the tissue of interest. Consequently, quantitative comparis on of any feature of interest typically uses a small fraction of the s ections from each retina, because analysis of the entire collection of sections is too time consuming. Such a sampling process can lead to m isleading or erroneous conclusions. We present a new method which allo ws complete analysis of the retina using a small number of samples pro duced by sectioning flattened retinas. This procedure is straightforwa rd as illustrated using an antibody against proliferating cell nuclear antigen (PCNA) to locate dividing cells in the teleost fish retina. I mmunocytochemical staining on flat-sectioned retinas was quantified us ing a computer-based image analysis system. When the cells of interest are randomly distributed, conventional sampling procedures can seriou sly under- or over-estimate their number. The new technique presented allows significantly more efficient examination and quantification of the entire retina as compared to conventional techniques.