As part of the California Current Ecosystem Long Term Ecological Research (CCE-LTER) Program, samples for epifluorescence microscopy and flow cytometry (FCM) were collected at ten 'cardinal' stations on the California Cooperative Oceanic Fisheries Investigations (CalCOFI) grid during 25 quarterly cruises from 2004 to 2010 to investigate the biomass, composition and size-structure of microbial communities within the southern CCE. Based on our results, we divided the region into offshore, and inshore northern and southern zones. Mixed-layer phytoplankton communities in the offshore had lower biomass (16±2μgCL-1; all errors represent the 95% confidence interval), smaller size-class cells and biomass was more stable over seasonal cycles. Offshore phytoplankton biomass peaked during the winter months. Mixed-layer phytoplankton communities in the northern and southern inshore zones had higher biomass (78±22 and 32±9μgCL-1, respectively), larger size-class cells and stronger seasonal biomass patterns. Inshore communities were often dominated by micro-size (20-200μm) diatoms; however, autotrophic dinoflagellates dominated during late 2005 to early 2006, corresponding to a year of delayed upwelling in the northern CCE. Biomass trends in mid and deep euphotic zone samples were similar to those seen in the mixed-layer, but with declining biomass with depth, especially for larger size classes in the inshore regions. Mixed-layer ratios of autotrophic carbon to chlorophyll a (AC:Chl a) had a mean value of 51.5±5.3. Variability of nitracline depth, bin-averaged AC:Chl a in the mixed-layer ranged from 40 to 80 and from 22 to 35 for the deep euphotic zone, both with significant positive relationships to nitracline depth. Total living microbial carbon, including auto- and heterotrophs, consistently comprised about half of particulate organic carbon (POC).