Characterization of the Mechanical Properties of Mouse Brain Tissue Using Atomic Force Microscopy

Abstract

Source: Canovic, E. P., et al. Characterizing Multiscale Mechanical Properties of Brain Tissue Using Atomic Force Microscopy, Impact Indentation, and Rheometry. J. Vis. Exp. (2016)

This video demonstrates the use of atomic force microscopy (AFM) to measure the viscoelastic properties of brain tissue, where a cantilever with a spherical probe interacts with the tissue and bends under applied forces. A laser beam deflected onto a detector tracks the cantilever to measure indentation depth and force relaxation, allowing for the characterization of tissue’s mechanical properties.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Atomic Force Microscope (AFM)-enabled Indentation

1. Prepare 60 mm-diameter Petri (P60) dishes with a mussel-derived bioadhesive according to the manufacturer's instructions.
   1. Prepare a stock of neutral buffer solution consisting of 0.1 M sodium bicarbonate in sterile water with an optimal pH of 8.0. Filter-sterilize (0.2 microns) the sodium bicarbonate buffer and store it at 4 °C.
   2. In a laminar flow hood, mix a solution of 6.25% mussel-derived bio-adhesive and 3.125% sodium hydroxid....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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