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Abstract Conjugated polymer (CP) aggregates have been the focus of considerable research, as these mesoscopic entities, compared with single CP chains, provide environments more analogous to those present in polymer-based optoelectronics in terms of the complexity of morphology and chain interactions; thereby, such aggregates hold the potential to provide insights into structure–function relationships highly relevant to optoelectronic device efficiency and stability. This review article highlights single-aggregate spectroscopy studies of CP aggregates based on a combination of solvent vapor annealing and single-molecule fluorescence techniques and draws mesoscopic connections between morphology, electronic coupling, and photophysics in CPs. This molecular-level understanding will pave the way for the bottom-up control of optoelectronic properties from the molecular to the device-length scale.