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Morphometry of the Respiratory Tract: Avoiding the Sampling, Size, Orientation, and Reference Traps

¹ California National Primate Research Center, University of California, Davis, CA 95616
² Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA 95616

Correspondence: Address correspondence to: Dallas M. Hyde, Director, California National Primate Research Center, One Shields Ave., University of California, Davis, CA 95616; e-mail:dmhyde{at}primate.ucdavis.edu

The extrapolation to humans of studies of infectious or toxic agents injurious to the respiratory system using animal models assumes comparability in the structure and function of animal models and humans. Measurement of conducting airways and parenchyma yields quantitative data for parameters like volume, surface area, length, cell number and cell size. Over the past few decades, there has been an evolution of rigorous uniform sampling designs of stereology that ensure unbiased estimates of number, length, surface area, and volume. This approach has been termed ‘design-based’ stereology because of the reliance on sampling design rather than geometric model-based stereology that makes assumptions. The aim of this paper is to define new design-based stereological approaches for the direct estimation of anatomical structures and epithelial, interstitial and endothelial cells of specific regions of the lung independent of the sampling, size, orientation and reference traps. An example is provided using wildtype and transgenic mice expressing transforming growth factor- to show the importance of the reference trap in stereologic estimates of postnatal lung growth.

Key Words: Stereology • dissector • fractionator • statistical variability • in vivo imaging • conducting airways

Abbreviations: IUR, Isotropic Uniform Random • SURS, Systematic, Uniformly Random Sampling • 3D, three-dimensional • V, Volume • S, Surface • L, Length • N, Number • Vv, Volume density or volume-to-volume ratio • Sv, Surface density or surface to volume ratio • Lv, Length density or length-to-volume ratio • Nv, Numerical density or number to volume ratio • I, Number of intersections • Q^–, Number of profiles or feature transects • t, Section thickness • p/l, Length per point on a test system • A, Area on a test system • h, Height of a dissector • bsf, Block sampling fraction • ssf, Section sampling fraction • asf, Area sampling fraction • hsf, Height sampling fraction • OCV²(), Observed coefficient of variation • CV²(X), Biological variation • CE²(), Sampling and stereological variation • TEM, Transmission Electron Microscopy

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