Home › Cell Biology › Micro-Mechanical Characterization of Lung Tissue Using Atomic Force Microscopy
Cell Biology JoVE (Open Access) Citable · DOI

Micro-Mechanical Characterization of Lung Tissue Using Atomic Force Microscopy

DOI: 10.3791/2911-v
What you'll learn
  • ✓ Prepare mouse lung tissue strips for mechanical characterization
  • ✓ Perform AFM microindentation on normal and fibrotic lung tissue
  • ✓ Extract and interpret stiffness data from AFM measurements
  • ✓ Correlate spatial variations in matrix stiffness with tissue pathology
Protocol

Biopharma Insights The stiffness of the extracellular matrix strongly influences multiple behaviors of adherent cells. Matrix stiffness varies spatially throughout a tissue, and undergoes modification in various disease conditions. Here we develop methods to characterize spatial variations in stiffness in normal and fibrotic mouse lung tissue using atomic force microscopy microindentation.

Difficulty
advanced
Total time
~3–4 hours per sample (tissue preparation through data acquisition)
Model organism
Mouse (lung tissue, presumed C57BL/6J or similar strain)
Biosafety
BSL-1

Steps

1
Prepare mouse lung tissue strips for analysis

Extract and prepare lung tissue sections from normal and fibrotic mouse lungs. Cut tissue into thin strips suitable for AFM microindentation, maintaining structural integrity and avoiding compression artifacts.

▶ 01:50
2
Mount tissue and perform AFM microindentation

Mount prepared lung tissue strips on AFM stage and conduct microindentation measurements at multiple spatial locations. Simultaneously acquire fluorescent imaging to identify tissue regions and correlate stiffness with cellular/structural features.

▶ 03:11
3
Extract and process AFM force-displacement data

Convert raw AFM force-displacement curves into quantitative stiffness measurements using appropriate mechanical models (e.g., Hertz contact mechanics). Generate stiffness maps and statistical summaries for normal versus fibrotic tissue.

▶ 07:14
4
Analyze and interpret representative AFM results

Review spatial stiffness distributions, compare mechanical properties between tissue types, and validate findings against morphological data. Discuss biological implications of matrix stiffness variations in normal and pathological lung.

▶ 08:48
💬 Comments coming soon