Protocol for generation, time-course imaging, and automated quality control of 3D spheroid invasion using TRACEQC

作者：Eric Cramer, Tamara Lopez-Vidal, Vania Wang, Jeanette Johnson, Daniel Bergman, Ashani T. Weeraratna, Elana J. Fertig, Laura M. Heiser, Young Hwan Chang, Jacquelyn W. Zimmerman · 发表于：STAR Protocols · 年份：2026 · DOI：10.1016/j.xpro.2026.104616 · 研究领域：Cell Image Analysis Techniques、Advanced Fluorescence Microscopy Techniques、Single-cell and spatial transcriptomics

Longitudinal 3D spheroid imaging requires quality control to ensure accurate image matching across sessions. We present a protocol for embedding spheroids and implementing TRACEQC (temporal reassignment and correspondence evaluation with quality control) to correct positional displacement. We describe steps for generating spheroids, embedding them in collagen plugs, acquiring image metadata, and applying TRACEQC to align spheroid positions across time points. This approach is agnostic to spheroid type, cellular composition, and formation technique, making it broadly applicable to longitudinal 3D imaging experiments. For complete details on the use and execution of this protocol, please refer to Cramer et al. 1