AUTHOR=Cheah Kevin , Chu Pingjin , Schmidt Greta , Scarlata Suzanne TITLE=Imaging methods to monitor and quantify cell differentiation JOURNAL=Frontiers in Cell and Developmental Biology VOLUME=Volume 13 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2025.1584858 DOI=10.3389/fcell.2025.1584858 ISSN=2296-634X ABSTRACT=The transition of a cell from a stem to a differentiated state involves an interrelated and complex series of events. These events include dynamic changes in cellular nucleic acid and protein content that are mediated by both intrinsic and extrinsic factors which ultimately lead to differentiation into specific lineage. Quantifying the parameters associated with differentiation and their changes under different conditions would not only allow for a better understanding of this process but also would enable the development of approaches that control differentiation. Here, we describe processes associated with the differentiation of two types of cultured cells, neurons and fibroblasts, and the tools to follow changes in real time. Specifically, we discuss methods to the identify cell lineage, changes in morphology, shifts in specific mRNA and miRNA levels as well as the changes in protein localization, interactions and assemblies that accompany differentiation.