Live Imaging of Cell Invasion Using a Multicellular Spheroid Model and Light-Sheet Microscopy

Area of Science:

• Cell Biology
• Microscopy
• Biotechnology

Background:

• Three-dimensional (3D) cellular assays are increasingly crucial for biological research, offering a more relevant model than traditional 2D cultures.
• Spheroids represent a key 3D model, serving as an essential intermediate step between 2D cell cultures and in vivo animal studies.
• Bridging the gap between in vitro and in vivo models is critical for accurate preclinical research and drug development.

Purpose of the Study:

• To present a straightforward protocol for generating and imaging spheroids.
• To demonstrate the application of light-sheet fluorescence microscopy for 3D cell assay analysis.
• To provide a reproducible method for sample preparation, image acquisition, and initial post-processing of spheroid data.

Main Methods:

• Detailed protocol for spheroid sample preparation.
• Image acquisition using the Zeiss Lightsheet Z.1 fluorescence microscopy platform.
• Initial post-processing steps for acquired spheroid images.

Main Results:

• Successful implementation of a user-friendly protocol for spheroid imaging.
• High-quality image data acquisition from 3D cellular spheroids.
• Demonstration of the utility of light-sheet microscopy for spheroid analysis.

Conclusions:

• The described protocol facilitates the use of spheroids as advanced 3D cell models.
• Light-sheet fluorescence microscopy provides an effective platform for imaging these complex biological structures.
• This method supports more accurate and predictive biological research by improving 3D model utilization.