Fluorescence Imaging of 3D Cell Models with Subcellular Resolution

Over the past years, research has made impressive breakthroughs towards the development and implementation of 3D cell models for a wide range of applications, such as drug development and testing, organogenesis, cancer biology, and personalized medicine. Opposed to 2D cell monolayer culture systems, advanced 3D cell models better represent the in vivo physiology. However, for these models to deliver scientific insights, appropriate investigation techniques are required. Despite the potential of fluorescence microscopy to visualize these models with high spatial resolution, sample preparation and imaging assays are not straightforward. Here, we provide different protocols of sample preparation for fluorescence imaging, for both matrix-embedded and matrix-free models (e.g., organoids and spheroids, respectively). Additionally, we provide detailed guidelines for imaging 3D cell models via confocal multi-photon fluorescence microscopy. We show that using these protocols, images of 3D cell culture systems can be obtained with sub-cellular resolution.

Automated summary

In recent years, there have been significant advances in the development and implementation of 3D cell models for various applications, including drug development, organogenesis, cancer biology, and personalized medicine. Unlike 2D cell culture systems, advanced 3D cell models provide a more realistic representation of in vivo physiology. However, appropriate investigation techniques are necessary for these models to deliver scientific insights. Fluorescence microscopy has the potential to visualize these models with high spatial resolution, but sample preparation and imaging assays can be challenging.