Quantitative Live-Cell Confocal Imaging of 3D Spheroids in a High-Throughput Format

Research Overview

Predicting human response to new compounds matters across many industries, yet standard in vitro and in vivo screening pipelines often lack predictive power. 3D cultures of human cells are more physiologically relevant and could support high-throughput automated testing of efficacy and toxicity — if the challenge of acquiring high-magnification confocal z-stacks of spheroids, and understanding their quantitative limits, can be solved.

The authors developed a method to form spheroids of reproducible size at precise spatial locations across a 96-well plate. Spheroids of varying radii were labeled with four fluorescent dyes and imaged on a high-throughput confocal microscope, and 3D renderings were analyzed to define what the imaging can and cannot quantify.