Meeting Abstract
Confocal laser scanning microscopy is an imaging technique that provides detailed optical sectioning of samples. We used this form of fluorescent microscopy to obtain three-dimensional images of harvestmen (“daddy-longlegs”) spermathecae, structures within the genitalia of female arthropods that store and maintain sperm after copulation. We examined spermathecal morphology in seven species of Leiobunum and one species of Hadrobunus, which were collected from North America and Japan and stored in 99% ethanol. Ovipositors were dissected and stained for 24 hours with DAPI to enhance the anatomical structures within the tissue. Although harvestmen spermathecae are naturally fluorescent, the use of DAPI was found to improve visualization of weakly sclerotized tissues. Z-stack images of the paired spermathecae were taken with a Leica SP5 confocal and three-dimensional representations were rendered in Imaris software using the contour surface tool. These are the first reported images of harvestmen spermatheca and we found that morphology varies among species, including differences in size, shape, and internal complexity. The method we developed allowed us to visualize internal structures difficult to interpret with two-dimensional brightfield microscopy, which could be applied to the characterization of internal structures in other arthropods.
