AbstractSediment flocculation is a critical component for the understanding of cohesive sediment dynamics. Traditionally, the referred study has largely been devoted to forming mechanism, influencing factors and physicochemical sediment conditions of all kinds of organic‐flocculation and inorganic‐flocculation. However, during the last decade, the bioflocculation of sediment by biological activity has been given increasing attention. But most studies have focused on the interrelations between biological and sedimentological variables. With the assistance of a newly developed field kit and correlative microscopy (which includes environmental scanning electron microscopy and confocal laser scanning microscopy), this article begins to bridge the resolution gap between sediment particles and biological activities as well as its metabolic products biofilm, in order to better understand the role of polymeric material biofilm in floc ultrastructure and outward floc behavior of bioflocculation sediment. Results have demonstrated that bioflocculation sediment was observed to be composed of complex networks of biofilm and appeared to be of complicated physical floc structures. The biofilm was found to embed particles and permeate the void space, representing the dominant physical bridging mechanism of the flocs and contributed to the extensive surface area, architecture characteristics, and mechanical properties of bioflocculation sediment. SCANNING 33:437–445, 2011. © 2011 Wiley Periodicals, Inc.