Related Experiment Video
Updated: Jun 13, 2026
Time-Resolved Fluorescence Imaging and Analysis of Cancer Cell Invasion in the 3D Spheroid Model
Published on: January 30, 2021
Protocol for generation, time-course imaging, and automated quality control of 3D spheroid invasion using TRACEQC
Eric M Cramer1, Tamara Lopez-Vidal2, Vania Wang3
1Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97239, USA; Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA.
Abstract:
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.
