The impact of experimental conditions on cell mechanics as measured with nanoindentation

Zambito, M., Viti, F., Bosio, A. G., Ceccherini, I., Florio, T. and Vassalli, M. (2023) The impact of experimental conditions on cell mechanics as measured with nanoindentation. Nanomaterials, 13(7), 1190. (doi: 10.3390/nano13071190)

Abstract

The evaluation of cell elasticity is becoming increasingly significant, since it is now known that it impacts physiological mechanisms, such as stem cell differentiation and embryogenesis, as well as pathological processes, such as cancer invasiveness and endothelial senescence. However, the results of single-cell mechanical measurements vary considerably, not only due to systematic instrumental errors but also due to the dynamic and non-homogenous nature of the sample. In this work, relying on Chiaro nanoindenter (Optics11Life), we characterized in depth the nanoindentation experimental procedure, in order to highlight whether and how experimental conditions could affect measurements of living cell stiffness. We demonstrated that the procedure can be quite insensitive to technical replicates and that several biological conditions, such as cell confluency, starvation and passage, significantly impact the results. Experiments should be designed to maximally avoid inhomogeneous scenarios to avoid divergences in the measured phenotype.

• Item Type: Articles
• Additional Information: This research was funded by the Italian Ministry of Health through “Ricerca Corrente” and “5 x mille” (to Gaslini Institute).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Vassalli, Professor Massimo
• Authors: Zambito, M., Viti, F., Bosio, A. G., Ceccherini, I., Florio, T., and Vassalli, M.
• College/School: College of Science and Engineering > School of Engineering > Biomedical Engineering
• Journal Name: Nanomaterials
• Publisher: MDPI
• ISSN: 2079-4991
• ISSN (Online): 2079-4991
• Published Online: 27 March 2023
• Copyright Holders: Copyright © 2023 The Authors
• First Published: First published in Nanomaterials 13(7): 1190
• Publisher Policy: Reproduced under a Creative Commons License
• Data DOI: 10.5281/zenodo.7766179