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Optimized murine lung preparation for detailed structural evaluation via micro-computed tomography.

Dragos M Vasilescu¹, Lars Knudsen, Matthias Ochs

¹Department of Radiology, University of Iowa, Iowa City, Iowa 52242, USA.

Journal of Applied Physiology (Bethesda, Md. : 1985)

|August 6, 2011

Summary

A new method uses micro-X-ray CT (μCT) to image mouse lungs, preserving their in vivo structure for detailed 3D analysis. This technique allows for stable, nondestructive ex vivo imaging of lung architecture at the alveolar level.

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Area of Science:

Pulmonary Imaging
Comparative Anatomy
Biomedical Engineering

Background:

Micro-X-ray computed tomography (μCT) offers high-resolution imaging of biological structures.
Visualizing intact lung architecture in three dimensions is crucial for understanding respiratory diseases.
Existing methods often struggle to preserve lung structure during ex vivo imaging.

Purpose of the Study:

To develop and validate a nondestructive fixation and imaging methodology for ex vivo micro-X-ray CT (μCT) of murine lungs.
To enable detailed three-dimensional morphometric analysis of lung architecture at the alveolar level.
To generate an atlas of in vivo and ex vivo lungs from normal murine strains.

Main Methods:

Developed a fixation protocol involving in situ vascular perfusion and air-drying of C57Bl/6 mouse lungs.

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Performed in vivo μCT imaging followed by ex vivo μCT imaging of fixed lungs at various time points.

Validated fixation stability by assessing lung volume changes over time (1 to >70 days).

Main Results:

The developed method successfully preserved lung air spaces and structural geometry for μCT imaging.
No significant lung shrinkage was observed post-fixation and air-drying, ensuring morphological stability.
Ex vivo μCT imaging revealed well-fixed alveoli and capillary beds, correlating with histological findings.

Conclusions:

A robust and nondestructive fixation and imaging method for ex vivo μCT of murine lungs has been established.
This technique allows for accurate ex vivo morphometric analysis representative of the in vivo lung state.
The method facilitates the creation of detailed 3D lung atlases for research.