Scanning electrochemical microscopy (SECM), a non-invasive variant of scanning probe microscopy, can be used to measure oxygen uptake or insulin secretion of isolated murine pancreatic islets in the presence of low and high physiological glucose concentrations levels. SECMimaging of islet topography and islet oxygen uptake is accomplished by the use of a redox mediator that cannot penetrate into the cell membranes of the islet coupled to direct reduction of oxygen at the SECM tip. An insulin sensing ultramicroelectrode (UME), adapted from a macro-scale insulin sensor incorporating a multiwalled carbon nanotube/dihydropyran film developed in our lab previously, was used for the direct electrochemical detection of insulin levels within extracellular media in real-time. We were able to stimulate and record increased insulin production in a single pancreatic islet by raising the concentration of extracellular glucose to the physiological 'high level.' These experiments generated probe scan curves (PSCs) for insulin production in the extracellular media surrounding single pancreatic islets extracted from male mice. This research is a part of an ongoing attempt to increase the scope of application for Bio-SECM, both through the adaptation of specialized biosensors and application of existing techniques for biological analysis of single cells and micro-organs. (C) The Author(s) 2015. Published by ECS. This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives 4.0 License (CC BY-NC-ND, http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is not changed in any way and is properly cited. For permission for commercial reuse, please email: [email protected]. [DOI: 10.1149/2.0111604jes] All rights reserved.