Semi-automated FIJI macro.

BackgroundChronic respiratory diseases represent a large group of non-communicable diseases that are a leading cause of mortality and morbidity globally. Many of the methods utilized to assess airway simplification in experimental models of the conditions are overly time-consuming and are sensitive to inter-operator biases, necessitating the need for unbiased and efficient tools to supplement analyses.MethodsWe propose a semi-automated method to quantitate the characteristics of large terminal respiratory airways and alveoli that uses free image-processing software (Fiji). We aimed to develop and test this method in a mouse model of bronchopulmonary dysplasia (BPD), a disease of blunted airway and pulmonary vascular development that remains a leading cause of mortality among preterm infants. Optimal macro parameters were determined with a test set of images from postnatal day 14 (P14) mice exposed to acute postnatal hyperoxia by determining which area and circularity values best correlated with mean linear intercept (LM). Validation was performed on a separate set of images from P7 mice subjected to the same hyperoxic model of BPD.ResultsBoth alveolar duct (r: 0.7866, p = 0.0359) and alveolar (r: 0.9475, p = 0.0012) area correlated with LM measurements from the test set. Using our method on a validation dataset, we demonstrate that hyperoxia-exposed mice possess fewer, enlarged alveoli that occupy less total area, as well as enlarged alveolar ducts that occupy a greater proportion of the parenchyma.ConclusionsWe report a semi-automated method of quantitating the characteristics of large and small terminal respiratory airways. This tool expedites analysis and removes operator bias relative to existing methods. We also demonstrate that LM changes in an acute model of hyperoxia-induced BPD result from both alveolar simplification and inadequate primary septation at the level of the alveolar ducts.