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Characterizing Multiscale Mechanical Properties of Brain Tissue Using Atomic Force Microscopy, Impact Indentation, and Rheometry

Published: September 6th, 2016



1Department of Materials Science and Engineering, Massachusetts Institute of Technology, 2Department of Biological Engineering, Massachusetts Institute of Technology, 3Department of Mechanical Engineering, Massachusetts Institute of Technology, 4Department of Neurology, The F.M. Kirby Neurobiology Center, Boston Children’s Hospital, Harvard Medical School

We present a set of techniques to characterize the viscoelastic mechanical properties of brain at the micro-, meso-, and macro-scales.

To design and engineer materials inspired by the properties of the brain, whether for mechanical simulants or for tissue regeneration studies, the brain tissue itself must be well characterized at various length and time scales. Like many biological tissues, brain tissue exhibits a complex, hierarchical structure. However, in contrast to most other tissues, brain is of very low mechanical stiffness, with Young's elastic moduli E on the order of 100s of Pa. This low stiffness can present challenges to experimental characterization of key mechanical properties. Here, we demonstrate several mechanical characterization techniques that have been adapted to measure the elastic and viscoelastic properties of hydrated, compliant biological materials such as brain tissue, at different length scales and loading rates. At the microscale, we conduct creep-compliance and force relaxation experiments using atomic force microscope-enabled indentation. At the mesoscale, we perform impact indentation experiments using a pendulum-based instrumented indenter. At the macroscale, we conduct parallel plate rheometry to quantify the frequency dependent shear elastic moduli. We also discuss the challenges and limitations associated with each method. Together these techniques enable an in-depth mechanical characterization of brain tissue that can be used to better understand the structure of brain and to engineer bio-inspired materials.

Most soft-tissues comprising biological organs are mechanically and structurally complex, of low stiffness compared to mineralized bone or engineered materials, and exhibit non-linear and time-dependent deformation. Compared to other tissues in the body, brain tissue is remarkably compliant, with elastic moduli E on the order of 100s of Pa 1. Brain tissue exhibits structural heterogeneity with distinct and interdigitated gray and white matter regions that also differ functionally. Understanding brain tissue mechanics will aid in the design of materials and computational models to mimic the response of the brain during injury, facilitate prediction ....

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Ethics Statement: All experimental protocols were approved by the Animal Research Committee of Boston Children's Hospital and comply with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

1. Mouse Brain Tissue Acquisition Procedures (for AFM-enabled indentation and impact indentation)

  1. Prepare a ketamine/xylazine mixture to anesthetize the mice. Combine 5 ml ketamine (500 mg/ml), 1 ml xylazine (20 mg/ml) and 7 ml of 0.9% saline solution.
  2. <.......

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Figure 4 shows representative indentation and force vs. time responses (Figure 4B,E) for creep compliance and force relaxation experiments, given an applied force or indentation depth (Figure 4A,D), respectively. Using these data and the geometry of the system, the creep compliance Jc(t) and force relaxation moduli GR(t) can be calculated for different regions of the brain (Figure 4C,F

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Each technique presented in this paper measures different facets of brain tissue's mechanical properties. Creep compliance and stress relaxation moduli are a measure of time-dependent mechanical properties. The storage and loss moduli represent rate-dependent mechanical properties. Impact indentation also measures rate-dependent mechanical properties, but in the context of energy dissipation. When characterizing tissue mechanical properties, both AFM-enabled indentation and rheology are commonly used methods. AFM-ena.......

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We acknowledge support of this work by the National Multiple Sclerosis Society and Simons Center for the Social Brain. BQ acknowledges support from the U.S. National Defense Science & Engineering Graduate Fellowship program.


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Name Company Catalog Number Comments
Xylaxine Lloyd Laboratoried perscription drug
Ketamine AnaSed Injections perscription drug
Vibratome (Vibrating blade microtome) Leica VT1200
Hibernate-A Medium Gibco A1247501 CO2-independent neural medium for adult tissue
Atomic Force Microscope, MFP-3D-BIO Asylum Research -
Petri Dish Heater Asylum Research -
AFM Probe, 0.03 N/m, 10 um radius borosilicate sphere Novascan PT.GS
Cell-Tak Corning 354240 mussel-derived bioadhesive
Sodium Bicarbonate Sigma-Aldrich S5761 alternate suppliers can be used
Sodium Hydroxide, 1N Sigma-Aldrich 59223C alternate suppliers can be used
Instrumented Indenter, NanoTest Vantage Micro Materials Ltd. - probe tip needs to be machined (steel flat punch, 1mm diameter, 4-5 mm length)
NanoTest Liquid Cell Micro Materials Ltd. -
Parallel Plate Rheometer MCR501 Anton-Parr -
PP25  Anton-Parr - 25 mm diameter flat measurement plate
Adhesive Sandpaper McMaster-Carr 4184A48 alternate suppliers can be used
Loctite 4013 Instant Adhesive Henkel 20268 alternate suppliers can be used

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