ABSTRACT The overall goal of this work is to experimentally observe stem cell differentiation, adipogenesis, of single cells to gain a more accurate model of the changes in mechanobiology over time. The high-speed simulation model of adipogenesis is currently based on observations of averaged ensemble properties of stem cell cultures. Because the model is of a single cell, its accuracy and predictive capability can be improved by obtaining more data on the changes in stem cell properties for single cells that can be tracked over time. An incubator will be developed as part of this grant to sustain stem cells under a microscope for at least 15 days, which will allow photographs at regular intervals of the morphology of the cell and nuclear membranes. However, time-dependent data on many cell properties is still difficult to obtain because the process of dyeing the cell to examine the state of its compo- nents, or probing its properties with an optical tweezer, typically kills the cell. Thus, the cells will be sacrificed after 15 days, and the high-speed simulation will be used work backward to estimate the initial properties related to successful and unsuccessful adipogenesis. Comparing successful and unsuccessful differentiation will yield biomarkers that provide early indication of successful adipogenesis, as proposed in Aim 1 of the parent grant. These additional single stem cell experiments will complement the data obtained from the ensemble experiments proposed in the parent grant.