A High-Throughput Platform for Assessing Single Fly Learning and Memory: Individual Drosophila Olfactory Conditioner (iDOC)

Olfactory conditioning in Drosophila melanogaster is a widely used behavioral paradigm for uncovering the neural basis of learning and memory. The traditional T-maze assay, however, relies on group-level assessments and cannot examine individual flies learning and memory abilities. To address this limitation, we developed the Individual Drosophila Olfactory Conditioner (iDOC), an apparatus for single-fly conditioning and memory assessment. Using iDOC, we demonstrated aversive olfactory learning, short-term memory (STM), anesthesia-resistant memory (ARM), and protein synthesis-dependent long-term memory (LTM) in individual flies, consistent with previous experiments conducted using the T-maze. As an individual memory assessment method, iDOC can be integrated with other behavioral assays at the single animal level, such as sleep monitoring. Notably, we found that spaced training, which induces LTM, also promotes post-learning sleep, while massed training, which induces ARM, did not. iDOC thus provides a powerful platform for investigating learning and memory and its interplay with other behavioral processes at the individual animal level. HighlightsO_LIDeveloped iDOC for high-throughput, single-fly aversive olfactory training and memory assessment C_LIO_LIDemonstrated aversive learning, STM, ARM, and LTM in individual flies using iDOC C_LIO_LISpaced conditioning, and not massed conditioning, promotes sleep C_LI