Images were captured using INFINITY series cameras (models 1-2C, 3-3URFC, and 2-1RC, Lumenera Corp., Ottawa, ON Canada) coupled with stereomicroscopes (model EMZ-TR, Meiji Techno, San Jose, CA USA and model M8, Wild Company, Heerbrugg, Switzerland) or a compound microscope (model Vanox, Olympus, Waltham, MA USA). Specimens >2mm in length were photographed superimposed over a 1mm gridded stage to provide spatial reference when comparing other individuals for verification. Standard photo perspectives (e.g., dorsal, ventral, lateral, etc.) were taken of each specimen, and at least one in each series was annotated to illustrate important family and/or genus level characteristics noted in commonly available dichotomous keys. Slide mounted specimens representing the family Chironomidae were photographed using a compound microscope, zoomed between 100X and 400X magnification. A similar set of standard perspectives were captured and annotated for these taxa.
The shallow depth of field associated with digital microscopy limits accurate representation of structures located at opposing focal lengths within a single image. To address this issue, we used ‘focus-stacking’ software Zerene Stacker (Zerene Systems) and INFINITY ANALYZE (Lumenera Corp.) to align and merge multiple images taken at variable focal planes to produce a single image that is completely in focus. Additionally, specimens or structures too large to capture in a single field of view were combined into a composite image using the open-source GNU Image Manipulation Program (GIMP, The GIMP Team, ver 2.8).