Hartsock, I.; Park, E.; Toppen, J.; Bubenik, P.; Dimitrova, E. S.; Kemp, M. L.; Cruz, D. A.

Understanding the multicellular organization of stem cells is vital for determining the mechanisms that coordinate cell fate decision-making during differentiation; these mechanisms range from neighbor-to-neighbor communication to tissue-level biochemical gradients. Current methods for quantifying multicellular patterning cannot capture the spatial properties of cell colonies across all scales and typically rely on human annotation or a priori selection of parameters. We present a computational pipeline that utilizes topological data analysis to generate quantitative, multiscale descriptors which capture the shape of data extracted from multichannel microscopy images. By applying our pipeline to certain stem cell colonies, we detected subtle differences in patterning that reflect distinct biological markers and progressive stages of differentiation. These results yield insight into directed cellular movement and morphogen-mediated, neighbor-to-neighbor signaling. Because of its broad applicability to immunofluorescence microscopy images, our pipeline is well-positioned to serve as a general-purpose tool for the quantitative study of multicellular pattern formation.