Organosulfate species have recently gained attention for their potentially significant contribution to secondary organic aerosol (SOA); however, their temporal behavior in the ambient atmosphere has not been probed in detail. In this work, organosulfates derived from isoprene were observed in single particle mass spectra in Atlanta, GA during the 2002 Aerosol Nucleation and Characterization Experiment (ANARChE) and the 2008 August Mini-Intensive Gas and Aerosol Study (AMIGAS). Real-time measurements revealed that the highest organosulfate concentrations occurred at night under a stable boundary layer, suggesting gas-to-particle partitioning and subsequent aqueous-phase processing of the organic precursors played key roles in their formation. Further analysis of the diurnal profile suggests possible contributions from multiple production mechanisms, including acid-catalysis and radical-initiation. This work highlights the potential for additional SOA formation pathways in biogenically influenced urban regions to enhance the organic aerosol burden.