Unbiased and efficient estimation of the total number of terminal bronchiolar duct endings in lung: a modified physical disector

A novel modification of the physical disector is described which was used t o estimate the total number of terminal bronchiolar duct endings (TBDEs) in human infant lung. TBDEs are closed three-dimensional space curves of comp lex shape that are inherently difficult to count from histological sections . However, careful consideration of the microanatomy of the terminal duct e ndings provides us with the opportunity to define a very simple and unbiase d counting rule. To apply the rule in practice we also need to determine a suitable disector height. Owing to the complex shape of the TBDE we had no prior knowledge of what disector height would be suitable for counting the TBDE structures. Exhaustive serial sectioning of complete TBDE structures w as carried out and showed that any disector height under 90 mu m would give unbiased counts. A further empirical study was then undertaken to determin e the most efficient disector height. This was found to be 50 mu m. The total number of TBDEs in the upper lobe of the right lung of six human infants aged between 13 and 25 weeks was also estimated. The estimates of n umerical density obtained with our modification of the physical disector we re multiplied by estimates of lung lobe volume obtained using Cavalieri's P rinciple. The total number of TBDEs in the lobes ranged from 15 323 to 57 7 68, with a mean of 40 306. The average coefficient of error of the number e stimates was 19%, which was deemed precise enough given the biological coef ficient of variation between TBDE number of 36%.