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Automated Image Analysis of Dictydium Sporothecae Biovolume Using Robotics

Dictydium cancellatum is a myxomycete (slime mold) of the order Liceales found mostly in wet, dark forest environments where it decays organic matter. A distinguishing feature of dictydium is the sporotheca, a bulbous container at the head of the organism that produces and contains the spore mass of the fruiting body, and as such plays an important reproductive role in the dictydium life cycle. Dictydium sporangia have been described as a cage, net, basket, or more commonly a Chinese lantern because of the unique shape created by thin strands of hyphae that make-up the peridial net. Biovolume measures cubic space of an object, and as such is one way to investigate changes to the sporotheca that could indicate a loss of reproductive viability of the organism, for example from stressors brought about by disruptions to ecological climate homeostasis. In this case study whole mount dictydium samples were used as a model to compare biovolume of the sporotheca for multiple specimens. This was accomplished by capturing digitalized microscopy images using a z-stacking method, whereby multiple images were taken at a constant focal height interval resulting in a depth value. A final z-projected image was then rendered from a range of in-focus z-stacks, and a primary mask was applied to the final region of interest to obtain length and width values. One way to study dictydium is by using a wet mount on standard microscopy slides, a format that can limit the efficiency of acquiring and analyzing large data sets. A robotic arm was therefore utilized to automate slide imaging at increased throughput, a solution that could enable larger population studies.