Confocal Imaging and Three-Dimensional Visualization of Thick Autofluorescent Specimens

Three-dimensional (3-D) rendering methods (maximum intensity projection, alpha blending, and isosurface rendering) are described for the visualization of thick, autofluorescent, arthropod cuticular structures (e.g., Drosophila melanogaster external genitalic structures) imaged by confocal laser scanning microscopy (CLSM). Additionally, specimen mounting and data collection strategies for thick specimens are described. Axial aberration artifacts are discussed in the context of these methods because of the critical roles they play in the quality of final 3-D images.