Traject3d allows label-free identification of distinct co-occurring phenotypes within 3D culture by live imaging

• DOI: 10.1038/s41467-022-32958-x
• Publication Date: 2022-09-09T17:19:04Z
• Authors: Eva C. Freckmann; Emma Sandilands; Erin Cumming; Matthew Neilson; Alvaro Román-Fern...; Konstantina Nikol...; Marisa Nacke; Tamsin R. M. Lann...; Ann Hedley; David Strachan; Mark Salji; Jennifer P. Morton; Lynn McGarry; Hing Y. Leung; Owen J. Sansom; Crispin J. Miller; David M. Bryant
• Keywords: Multicellular organism; Identification; Live cell imaging; 3D cell culture; Phenotypic screening

Abstract Single cell profiling by genetic, proteomic and imaging methods has expanded the ability to identify programmes regulating distinct states. The 3-dimensional (3D) culture of cells or tissue fragments provides a system study how such states contribute multicellular morphogenesis. Whether plated into 3D cultures give rise singular phenotype whether multiple biologically phenotypes arise in parallel is largely unknown due lack tools detect heterogeneity. Here we develop Traject3d (Trajectory identification 3D), method for identifying heterogeneous these over time, from label-free multi-day time-lapse imaging. We use this characterise temporal landscape morphological cancer lines, varying metastatic potential drug resistance, information combinations that inhibit therefore an important companion other single-cell technologies facilitating real-time via live can lead alternate occur culture.