Abstract The Nebraska Center for Cellular Signaling (NCCS) Live-Cell Imaging Core provides a variety of support mechanisms to investigators within the NCCS and our partnering institutions through consultation services, reagents, and expertise in an array of microscopy techniques. The original goal of making advanced microscopy technologies readily available to the members of the NCCS continues to drive the success of the core and has been extended to support the larger research community. Since its formation at the beginning of Phase II, the Live-Cell Imaging Core has supported investigators through the generation of data for grant submissions and publications. The Live-Cell Imaging Core is composed of facilities with shared live-cell imaging capabilities as well as specialized features for unique applications. As part of Phase III funding of the NCCS, we are requesting funds to support the aims of the Live-Cell Imaging Core. Aim 1: Provide and maintain state-of-the-art live-cell microscopy equipment and offer technical support to investigators both within and outside the NCCS. Aim 2: Assist in the generation of image acquisition and analysis, allowing investigators to incorporate live-cell microscopy into their thinking and into their projects and increasing the likelihood grant proposals will be funded. Aim 3: Expand the capacity for collaboration between our Center and its partnering institutions. During Phase II, the Live-Cell Imaging Core established the technologies and the systems needed for advanced microscopy platforms. Together with strong institutional support, we propose to continue our momentum by implementing super-resolution microscopy capabilities for TIRF (Total Internal Reflection Fluorescence) microscopy and PALM (Photoactivated Localization Microscopy) within the Live-Cell Imaging Core. Addition of these technologies will greatly enhance the ability of NCCS investigators to compete for extramural funding. Long-term sustainability of the Live-Cell Imaging Core will be achieved by: 1) maintaining a strong base of well-funded users; 2) expansion of collaborations with users outside the NCCS; and 3) continuation of steady institutional support.