Genetically encoded fluorescent and bioluminescent reporters are now widely used for imaging and understanding of intracellular signaling in response to extracellular stimuli in real time in single living cells. Primary cultures are a valuable tool, and are often preferred over transformed or immortalized cell lines, since they are biologically more relevant and important in biomedical research and therapeutic development. To incorporate genetically encoded reporters into the primary culture of non-dividing cells, such as mouse or human pancreatic acinar cells, is not an easy task. This short review discusses the different methods available to transfect cell lines and primary cultures while especially focusing on pancreatic acinar cells with genetically encoded optical reporters to advance knowledge of the pathophysiology of pancreatitis.