Motivation: Shifts in nutritional habits and relative abundance of refined high-calorie foods in modern postindustrial societies resulted in increased prevalence of both type I and type II diabetes mellitus, causing significant morbidity and mortality and incurring increasingly higher healthcare and economic costs. A promising therapy is autologous transplantation of insulin-secreting b cells obtained via redifferentiation of patient’s own induced pluripotent stem cells (iPSC) (1), including for juvenile diabetes supplementary means to prevent autoimmune attack by effector T cells (2). The complex insulin secretion machinery comprises the interplay of several ion channels: cell depolarization via ATP inhibition of inward rectifier K(ATP) channels upon glucose entry is followed by opening of voltage-dependent Ca2+ channels triggering exocytosis of insulin (3). The assessment of these ion channels in b cells or correspondent cell lines applying automated patch-clamp provides an efficient tool for analyzing these processes and identifying therapeutic strategies. Methods: Whole-cell patch-clamp experiments were performed in the rat insulinoma cell line INS-1 (4), kindly provided by Prof. Baltrusch and Dr. Schultz from the Institut für Medizinische Biochemie, Universität Rostock, using the CytoPatch™ equipment (5), with dual-channel quartz pipette tips 2 mm in diameter. Depolarizing voltage steps (-70 to +40 mV for 300 ms) or ramps from -100 to +100 mV were applied regularly, while K+ concentration was increased from 2.5 to 20 mM. The internal solution contained either 5 mM ATP or was ATP-free. Results: Automated whole-cell recordings utilizing the INS-1 cells have been established. Depolarizing steps elicited large delayed rectifier and transient outward currents, which were strongly inhibited by either 4-aminopyridine 1 mM or tetraethyl ammonium 5 mM. With ATP-free internal solution we recorded large inward rectifying currents, displayed as difference of traces at 20 mM and 2.5 mM external K+, amounting to -200 pA/pF at -100 mV, with a reversal potential close to the K+ equilibrium potential in the specified ionic conditions. Conclusion: Automated patch-clamp technology utilizing the CytoPatch™ can be successfully used via whole-cell experiments to investigate the signaling pathways and ionic currents involved in insulin secretion. It is therefore an appropriate tool to be applied for quality control and validation of iPSC-derived pancreatic b cells in view of clinical applications and to screen against modulators of those currents for pharmacological purposes.