Novel methodology utilizing confocal laser scanning microscopy for systematic analysis in arthropods (Insecta)

The use of confocal laser scanning microscopy (CLSM) for imaging arthropod structures has the potential to profoundly impact the systematics of this group. Three-dimensional visualization of CLSM data provides high-fidelity, detailed images of minuscule structures unobtainable by traditional methods (for example, hand illustration, bright-field light microscopy, scanning electron microscopy). A CLSM data set consists of a stack of 2-D images (optical slices) collected from a transparent, fluorescent specimen of suitable thickness. Small arthropod structures are particularly well suited for CLSM imaging owing to the autofluorescent nature of their tissues. Here, we document the practical aspects of a methodology developed for obtaining image stacks via CLSM from autofluorescent insect cuticular structures.