The Drosophila Shallow Chamber (DSC) is an essential tool for examining the locomotor behavior of individual Drosophila melanogaster flies, interactions between pairs, or complex social interactions within large groups. Over the years, studies on Drosophila have provided significant insights into the molecular mechanisms governing behavior and development. Despite differences in morphology and cellular features, Drosophila melanogaster shares many functionally analogous internal organ systems with vertebrates, including humans. Historically, the morphological differences limited the use of invertebrate organs in studying pathological mechanisms. However, advancements in genomic mapping have revealed that approximately 65% of human disease-causing genes have functional homologs in flies, which express proteins with similar functions to those in humans. This discovery has been pivotal in identifying molecular mechanisms underlying mutations in the human genome that lead to disease phenotypes, particularly those affecting the nervous system. Drosophila, with its smaller size and less complex nervous system compared to many vertebrates, serves as a model for studying various neurological diseases, such as neurodegeneration, epilepsy, dementia, stroke, traumatic brain injury, and brain tumors. These studies offer novel insights into pathogenic mechanisms and aid in developing therapeutic strategies for neurological disorders (Ugur et al., 2016).

While numerous genetic tools exist to study the relationships between genes, neurons, and behavior in flies, the development of tools for quantifying behavior has lagged, slowing progress in understanding these relationships. Drosophila behavior is often studied through digital recordings using machine vision methodologies that automatically track and measure behavioral phenotypes. The effectiveness of these methodologies depends heavily on the quality of raw data recorded from experimental chambers. Traditional chambers with vertical walls pose significant challenges for behavioral measurement and analysis. Vertical walls enable flies to walk up and along the ceiling, causing overlapping and blocking behaviors, which complicates individual tracking. This movement can obscure key identifiable features such as the position of wings or limbs, critical for analyzing specific behaviors. High ceilings allow ample room for flight and space for flies to cluster in the periphery, making tracking difficult with a single camera. To address these issues, intricate designs incorporating thermal barriers and water moats were developed (Simon et al., 2010). These designs often required wing clipping to prevent flight, potentially interfering with behaviors such as courtship and aggression. Simon et al. introduced a simplified experimental chamber (DSC) with a low ceiling and sloping walls, creating a shallow space that forces flies to form a monolayer, preventing overlap and appearance changes. This chamber complements various machine vision methodologies for behavioral measurement and analysis using an overhead camera.

The Drosophila Shallow Chamber is a versatile experimental tool featuring sloping walls rather than vertical ones with square corners. The sloping walls intersect with a paint-coated glass ceiling and are continuous with the chamber’s horizontal floor. The floor has holes for introducing food and water. The chamber is illuminated from above with fluorescent lights and from below with infrared LED lights. A camera positioned above the chamber records fly behavior.