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Am J Physiol Lung Cell Mol Physiol (September 13, 2002). doi:10.1152/ajplung.00184.2002
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Articles in PresS, published online ahead of print September 13, 2002
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00184.2002
Submitted on June 10, 2002
Accepted on August 30, 2002

Variation of lung volume after fixation measured by immersion or Cavalieri method

Xiao Yan1, Juan Jose Polo Carbayo1, Ewald R. Weibel2, and Connie C. H. Hsia1*

1 Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX, USA
2 Institute of Anatomy, University of Bern, Bern, Switzerland

* To whom correspondence should be addressed. E-mail: Connie.Hsia{at}utsouthwestern.edu.

Organ volume is a critical parameter in morphometric analysis. The special problems of the lung as a non-solid organ are overcome by tracheal instillation of fixatives at a constant airway pressure (Paw). Lung volume can change significantly after fixation as Paw change. To determine the variation of lung volume after fixation, we measured the volume of intact fixed lungs by serial immersion in saline (Vimm) at selected time points, in comparison to measurements obtained by point counting (Cavalieri Principle, Vcav) after tissue sectioning to release Paw. Vimm was systematically higher than Vcav, by 25% in dog lungs and 13% in guinea pig lungs (p=0.0003 between species). This size-dependent variability reflects residual elastic recoil, refolding and/or crumpling of alveolar septa after fixation. Vimm remained 14% higher than Vcav in dog lungs even after pressure release. Vcav/Vimm was systematically lower in the upper than the lower strata of the same lung. We conclude that Vcav measured on lung slices after relaxation of Paw more precisely represents the state of the tissue to be used for subsequent morphometric analysis, particularly for large lungs.




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