A Low‐Cost, User‐Friendly Rheo‐Optical Compression Assay to Measure Mechanical Properties of Cell Spheroids in Standard Cell Culture Plates

Abstract

The mechanical characterization of cell spheroids, one of the most widely used 3D biology models in vitro, is a hotspot of current research on the role played by the mechanical response of cells and tissues. The techniques proposed so far in the literature, while providing important scientific insights, require specialized equipment and technical skills that are not usually available in cell biology facilities. Here, an innovative rheo‐optical compression assay is presented: it is based on microscopy glass coverslips as the load is applied to cell spheroids in standard cell culture plates and on image acquisition with an optical microscope or even a smartphone equipped with adequate magnification lenses. Mechanical properties can be simply obtained by correlating the applied load to the deformation of cell spheroids measured by image analysis. The low‐cost, user‐friendly features of the proposed technique can boost mechanobiology research making it easily affordable to any biomedical lab equipped with cell culture facilities.

i-Rheo-optical assay: Measuring the viscoelastic properties of multicellular spheroids

“…In particular, the cell spheroids were collected from the 48-well plate by using a pipette, and then placed into a 6-well plate containing approximately 3 mL of phosphate-buffered solution (PBS); thus, ensuring no wall effects and a consistent settling of the glass coverslip. For more information on the above process, the reader is referred to our recent publication [ 41 ].…”

“…The liquid overlay technique was employed for spheroids preparation, selected for its judicious balance of cost-efficiency and ease of management [ 39 , 40 ]. At the core of our rheo-optical compression assay was the utilization of a microscope coverslip to apply a constant load, resulting in the development of a cost-effective and user-friendly technique [ 41 ]. Essentially, our innovative approach produces strain-time curves of pivotal significance in elucidating the mechanical properties of the spheroids.…”