Experimental and data analysis workflow for soft matter nanoindentation

Ciccone, G., Azevedo Gonzalez Oliva, M., Antonovaite, N., Lüchtefeld, I., Salmeron-Sanchez, M. and Vassalli, M. (2022) Experimental and data analysis workflow for soft matter nanoindentation. Journal of Visualized Experiments, 179, e63401. (doi: 10.3791/63401)

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Abstract

Nanoindentation refers to a class of experimental techniques where a micrometric force probe is used to quantify the local mechanical properties of soft biomaterials and cells. This approach has gained a central role in the fields of mechanobiology, biomaterials design and tissue engineering, to obtain a proper mechanical characterization of soft materials with a resolution comparable to the size of single cells (μm). The most popular strategy to acquire such experimental data is to employ an atomic force microscope (AFM); while this instrument offers an unprecedented resolution in force (down to pN) and space (sub-nm), its usability is often limited by its complexity that prevents routine measurements of integral indicators of mechanical properties, such as Young's Modulus (E). A new generation of nanoindenters, such as those based on optical fiber sensing technology, has recently gained popularity for its ease of integration while allowing to apply sub-nN forces with µm spatial resolution, therefore being suitable to probe local mechanical properties of hydrogels and cells. In this protocol, a step-by-step guide detailing the experimental procedure to acquire nanoindentation data on hydrogels and cells using a commercially available ferrule-top optical fiber sensing nanoindenter is presented. Whereas some steps are specific to the instrument used herein, the proposed protocol can be taken as a guide for other nanoindentation devices, granted some steps are adapted according to the manufacturer's guidelines. Further, a new open-source Python software equipped with a user-friendly graphical user interface for the analysis of nanoindentation data is presented, which allows for screening of incorrectly acquired curves, data filtering, computation of the contact point through different numerical procedures, the conventional computation of E, as well as a more advanced analysis particularly suited for single-cell nanoindentation data.

Item Type:Articles
Additional Information:GC and MAGO acknowledge all members of the CeMi. MSS acknowledges support via an EPSRC Programme Grant (EP/P001114/1).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Salmeron-Sanchez, Professor Manuel and Vassalli, Dr Massimo and Azevedo Gonzalez Oliva, Mariana and CICCONE, Mr GIUSEPPE
Creator Roles:
Ciccone, G.Software, Formal analysis, Validation, Investigation, Data curation, Writing – original draft, Writing – review and editing, Visualization
Azevedo Gonzalez Oliva, M.Investigation, Writing – original draft, Writing – review and editing, Visualization
Salmeron-Sanchez, M.Resources, Writing – original draft, Writing – review and editing, Supervision, Project administration, Funding acquisition
Vassalli, M.Conceptualization, Software, Validation, Resources, Writing – original draft, Writing – review and editing, Supervision, Project administration, Funding acquisition
Authors: Ciccone, G., Azevedo Gonzalez Oliva, M., Antonovaite, N., Lüchtefeld, I., Salmeron-Sanchez, M., and Vassalli, M.
College/School:College of Science and Engineering
College of Science and Engineering > School of Engineering
College of Science and Engineering > School of Engineering > Biomedical Engineering
Journal Name:Journal of Visualized Experiments
Publisher:JoVE
ISSN:1940-087X
ISSN (Online):1940-087X
Published Online:18 January 2022
Copyright Holders:Copyright Copyright © 2022 JoVE
First Published:First published in Journal of Visualized Experiments 179: e63401
Publisher Policy:Reproduced under a Creative Commons licence

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
173192Engineering growth factor microenvironments- a new therapeutic paradigm for regenerative medicineManuel Salmeron-SanchezEngineering and Physical Sciences Research Council (EPSRC)EP/P001114/1ENG - Biomedical Engineering