Comparing Automated Morphology Quantification Software on Dendrites of Uninjured and Injured Drosophila Neurons.

Dendrites shape inputs and integration of depolarization that controls neuronal activity in the nervous system. Neuron pathologies can damage dendrite architecture and cause abnormalities in morphologies after injury. Dendrite regeneration can be quantified by various parameters, including total dendrite length and number of dendrite branches using manual or automated image analysis approaches. However, manual quantification is tedious and time consuming and automated approaches are often trained using wildtype neurons, making them poorly suited for analysis of genetically manipulated or injured dendrite arbors. In this study, we tested how well automated image analysis software performed on class IV Drosophila neurons, which have several hundred individual dendrite branches. We applied each software to automatically quantify features of uninjured neurons and neurons that regenerated new dendrites after injury. Regenerated arbors exhibit defects across multiple features of dendrite morphology, which makes them challenging for automated pipelines to analyze. We compared the performances of three automated pipelines against manual quantification using Simple Neurite Tracer in ImageJ: one that is commercially available (Imaris) and two developed by independent research groups (DeTerm and Tireless Tracing Genie). Out of the three software tested, we determined that Imaris is the most efficient at reconstructing dendrite architecture, but does not accurately measure total dendrite length even after intensive manual editing. Imaris outperforms both DeTerm and Tireless Tracing Genie for counting dendrite branches, and is better able to recreate previous conclusions from this same dataset. This thorough comparison of strengths and weaknesses of each software demonstrates their utility for analyzing regenerated neuron phenotypes in future studies.