The most frequent causes of acute pancreatitis are either gallstones or heavy alcohol consumption. Recurrent episodes of acute pancreatitis often result in progression of the disease to chronic pancreatitis and increase the risk of pancreatic cancer. The pathogenesis of inflammation in acute and chronic pancreatitis is tightly linked to the induction of necrosis and apoptosis. There is currently no specific therapy for pancreatitis; however, latest findings allow hope for future progress. Single pancreatic acinar cells and clusters of two or three acinar cells were isolated from the mouse pancreas. The mice were humanely killed according to the UK Schedule 1 regulation of the Animal (Scientific Procedures) Act, 1986. Cells were loaded with calcium sensitive dye Fluo-5N AM followed by permeabilization using a short pulse of high intensity two-photon laser beam. BZiPAR was used to continuously measure the activity of trypsin by adding the substrate to the perfusion chamber after cell permeabilization. Measurements of fluorescence were performed using Leica multi-photon SP5 system. Recently we have shown that the calcium sensor calmodulin provides a protective mechanism, regulating the sensitivity of the calcium release process. Activation of calmodulin using Ca2+-like peptide 3 (CALP-3) abolished the pathological calcium release and trypsinogen activation evoked by ethanol as well as its non-oxidative metabolite palmitoleic acid ethyl ester (POAEE) in intact and permeabilized cells. We have experimentally compared a number of ways to reduce calcium overload inflicted by known pancreatitis inducing factors (alcohol, fatty acid ethyl esters, bile acids, high concentration of cholecystokinin) including inhibition of calcium release, inhibition of calcium entry, potentiation of calcium recovery and activation of calcium regulation mechanisms. Our data suggest that activation of calcium regulation and inhibition of calcium entry (or possibly combination of both) reduces calcium overload as well as necrosis, bringing hope for new therapeutic approaches.