Neuronal synchronization contributes to various cognitive functions and disruption in synchronicity may lead to various diseased conditions. However, measurement of synchronicity at a higher spatial resolution remains challenging. Specifically, investigation on understanding the role of network topology in tuning the network activity and synchronicity remains sparse. In this context, we propose imaging of intracellular Ca2+ in primary cultures of hippocampal neurons using Fluo-4 as the fluorescent indicator using the confocal microscope. In order to identify the synchronous response from a set of heterogeneous Ca2+ spiking, we present fuzzy clustering of the oscillatory responses. Further, the synchronicity was measured through evaluation of the correlation between Ca2+ spiking trends. Confocal imaging and analysis show that neuronal connectivity and topology play an essential role in tuning the synchronicity of the neuronal network. © 2018 IEEE.