Utility of fluorescence microscopy in embryonic/fetal topographical analysis

Abstract

For topographical analysis of developing embryos, investigators typically rely on scanning electron microcopy (SEM) to provide the surface details not attainable with light microscopySEM is an expensive and time consuming technique, however, and the preparation procedure may alter morphology and leave the specimen friable. We report that by using a high‐resolution compound epilfluorescence microscope with inexpensive low‐power objectives and the flurochrome acridine orange, we were able to obtain surface image of fixed or fresh whole rat embryos and fetal palates of considerably greater topographical detail than those obtained using routine light microscopy. Indeed the resulting high‐resolution image offered not only superior qualitative documentation of morphological observation, but the capability for detailed morphometry via digitization and computer‐assisted image analysis.