This is an administrative supplement application for purchasing a stage top incubator to replace the malfunctioning enclosure-style incubator installed on the confocal-TIRF microscope system in the PI’s lab more than 15 years ago. A functioning incubator that provides optimal temperature, CO2, and humidity to keep cells healthy during imaging experiments is critical for the success of the parent project that heavily relies on live-cell imaging assays to define mechanisms regulating PIP2 homeostasis by Nir2 protein at endoplasmic reticulum (ER)-plasma membrane (PM) junctions during receptor-induced Ca2+ signaling. The purchase of a new stage top incubator system is urgently needed to reinstate live cell imaging capability of proposed experiments crucial for the success of the parent project. A new stage top incubator system will provide a consistent environment for cells to stay healthy for high-quality and quantitative imaging results which is critical for the awarded project.