The appropriate activation of B cells is critical for the development and operation of immune responses and is dependent on the extensive coordination of intra- and intercellular communications in response to antigen stimulation. An accurate description of the B cell-activation process requires investigation of these interactions within their correct cellular context both at high resolution and in real time. Here, we discuss a number of recent studies that have offered insight into the early molecular events of B cell activation. We suggest that segregation within the B cell membrane triggers localized cytoskeleton reorganisation and signaling, allowing the formation of B cell receptor (BCR) microclusters. These BCR microclusters are the sites for the coordinated recruitment of the signalosome and are propagated during B cell spreading. We discuss the recent identification of a critical role for CD19 in the B cell response to membrane-bound antigen and suggest a mechanism involving BCR microclusters by which it mediates its stimulatory function. Finally, we consider research that has taken advantage of recent technological advances in multiphoton microscopy that have allowed its application to the investigation of the dynamics of membrane-bound antigen presentation and subsequent B cell activation in lymph nodes in vivo.