Morphological studies in spider and insect groups (Phylum Arthropoda) have historically relied upon traditional imaging techniques such as light microscopy (LM) and scanning electron microscopy (SEM). It has often been difficult, however, to examine some structural details of certain specimens due to limitations of the techniques. Light microscopy does not always provide adequate depth of focus for visualizing the true three-dimensional structure of a specimen, and out-of-focus light in the image can obscure detail. SEM is not applicable if the specimen is too delicate to undergo critical point drying. Additionally, SEM is a surface technique and does not provide any information about internal structure.
If a specimen is transparent to light, information about the morphology of internal structures, and connections between structures, can easily be obtained using confocal microscopy without the problems associated with traditional LM. In this study, we used confocal laser scanning microscopy (CLSM) to study the morphology of chitinous arthropod structures by taking advantage of their autofluorescent properties.