Recent advancements in biomedical imaging focus on fluorescent imaging using laser scanning confocal microscopy. However, high-resolution imaging of cellular activity remains considerably expensive for both in vitro and in vivo model. In this context, integration of mathematical modeling and imaging data analysis to predict the cellular activity may aid understanding of cell signaling. Here we performed dynamic imaging using confocal microscopy and propose a model considering cell to cell connectivity that can predict the effect of the drug on Ca2+ oscillations. The proposed model consists of a large number of ordinary differential (ODE) equations and uses the concept of adjoint matrix containing coupling factors to capture the activity of cells with the random arrangement. The results show that the cell-to-cell connection plays a crucial role in controlling the calcium oscillations through a diffusion-based mechanism. The present simulation tool can be used as a generalized framework to generate and visualize the time-lapse videos required for in vitro drug testing for various drug doses. © 2020 IEEE.