Chronic Hypoxia and Hyperoxia Modifies Morphology and Vegf Expression of the Lungs of the Developing Chicken (Gallus Gallus Domesticus) Page: 27
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parabronchial tissue, as well as the most homogenous in nature, when using entire
cross-sections for analysis and comparison. Each section was photographed using an
Olympus DP70 digital camera system mounted on to a light microscope. All sections were
photographed at a total magnification of 40x. The following stereological measurements
were taken for each section, utilizing ImageJ (National Institutes of Health).
Lung Surface Areas
Total Lung Surface Area
Surface areas for each cross-section were calculated in 2-dimensions,
measuring the X and Y axes. Total surface area for each cross-section was determined
by tracing the edges of the entire section. The total surface area for each section then
served as a reference by which to determine the fractional percentage (proportion) of
lumina, parabronchial tissue, and non-parabronchial tissue to total lung tissue.
Although, shrinkage occurs in pulmonary tissues due to histological processing, the
components of the exchange tissue in avian lungs shrink in relative proportions (Maina
et al., 1989). Therefore, the fixation methods preserved the proportional differences
between components, samples, and groups.
The components of total surface area that were measured were total lumina, total
parabronchial tissue, parabronchi with lumina, and non-parabronchial tissue. The
proportion of each component was calculated using the following formula:
Component % = SAcomp / SAlung
where SAlung is an estimate of the total surface area of the lung section by tracing
around the entire outside edge of the lung section, and SAcomp is an estimate of
the surface area of the component within the entire lung section by tracing
around all occurrences of that component within the section.27
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Lewallen, Melissa Anjanette. Chronic Hypoxia and Hyperoxia Modifies Morphology and Vegf Expression of the Lungs of the Developing Chicken (Gallus Gallus Domesticus), thesis, December 2012; Denton, Texas. (https://digital.library.unt.edu/ark:/67531/metadc177224/m1/34/: accessed April 19, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; .