ArticleNature protocols2021
Measuring the elastic modulus of soft culture surfaces and three-dimensional hydrogels using atomic force microscopy.
Article in Nature protocols, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 72 papers.
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Who cites it
72 citing papers in PubMed, 151 citations in OpenAlex.
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- Simple fabrication method for cancer cell migration studies on biomimetic substrates with tunable stiffness.Materials today. Bio · 2026Article
- Mechanobiology of the tumor microenvironment: a review of therapeutic interactions and in vitro elasticity measurement techniques.Journal of biomedical science · 2026Review
- Frequency-dependent cellular microrheology with pyramidal atomic force microscopy probes.Biophysical journal · 2026Article
- Microscale Mechanical Cues in Hydrogels: Engineering Strategies to Modulate Cell Fates in Three Dimensions.Cell biomaterials · 2026Article
- Microengineered Gradient Hydrogels for Mechanobiology.Advanced healthcare materials · 2026Review
- Fluorescently Labeled Gradient Hydrogels Reveal Matrix-Dependent Cell Responses to Substrate Stiffness.Small (Weinheim an der Bergstrasse, Germany) · 2026Article
- Mechanotyping of Organoids for Assessing Drug-Induced Injuries.Advanced materials (Deerfield Beach, Fla.) · 2026Article
- Bioengineering Stem Cell-Derived Glioblastoma Organoids: A Comprehensive Review.Pharmaceuticals (Basel, Switzerland) · 2025Review
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- Analysis of the Hertz Contact Model for Evaluating Mechanical Properties of Polymers Using the Finite Element Method.Polymers · 2025Article
- Atomic Force Microscopy for Cross-Disciplinary Materials Research.Small methods · 2025Review
- Targeting Fibrotic Scarring by Mechanoregulation of Il11ra1Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Mechanical Properties of Endothelial Cells: A Key to Physiology, Drug Testing and Nanostructure Interaction.Cells · 2025Review
- The role of non-linear viscoelastic hydrogel mechanics in cell culture and transduction.Materials today. Bio · 2025Review
- An Organ-on-Chip Platform for Strain-Controlled, Tissue-Specific Compression of Cartilage and Mineralized Osteochondral Interface to Study Mechanical Overloading in Osteoarthritis.Advanced healthcare materials · 2025Article
- High-Throughput Mechanical Characterization of Single Microgel Particles by Fluidic Force Microscopy.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- 3D bioprinting of a perfusable skin-on-chip model suitable for drug testing and wound healing studies.Materials today. Bio · 2025Article
- Elastic modulus of hyaluronic acid hydrogels by compression testing.Journal of materials science. Materials in medicine · 2025Article
12 more citing papers are in PubMed but not listed here.
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
Abstract
Growing interest in exploring mechanically mediated biological phenomena has resulted in cell culture substrates and 3D matrices with variable stiffnesses becoming standard tools in biology labs. However, correlating stiffness with biological outcomes and comparing results between research groups is hampered by variability in the methods used to determine Young's (elastic) modulus, E, and by the inaccessibility of relevant mechanical engineering protocols to most biology labs. Here, we describe a protocol for measuring E of soft 2D surfaces and 3D hydrogels using atomic force microscopy (AFM) force spectroscopy. We provide instructions for preparing hydrogels with and without encapsulated live cells, and provide a method for mounting samples within the AFM. We also provide details on how to calibrate the instrument, and give step-by-step instructions for collecting force-displacement curves in both manual and automatic modes (stiffness mapping). We then provide details on how to apply either the Hertz or the Oliver-Pharr model to calculate E, and give additional instructions to aid the user in plotting data distributions and carrying out statistical analyses. We also provide instructions for inferring differential matrix remodeling activity in hydrogels containing encapsulated single cells or organoids. Our protocol is suitable for probing a range of synthetic and naturally derived polymeric hydrogels such as polyethylene glycol, polyacrylamide, hyaluronic acid, collagen, or Matrigel. Although sample preparation timings will vary, a user with introductory training to AFM will be able to use this protocol to characterize the mechanical properties of two to six soft surfaces or 3D hydrogels in a single day.
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