Estimation of podocyte number: A comparison of methods

Background. The podocyte is the focus of much research into the mechanisms of renal disease progression, and the number of podocytes per glomerulus has thus become a parameter of much interest. When counting podocytes, the actual particle counted is the cell nucleus. The majority of published studies estimating podocyte number have used the method of Weibel and Gomez (1962). This makes assumptions about the shape and size of the cell nuclei and therefore has an inherent bias. In our studies we have used a more recent stereologic method-the disector/fractionator- that makes no assumptions about the shape or size of the cell nuclei and is therefore free of bias. Methods. We set out to compare the two methods, in both type 1 diabetic patients and normal controls, to determine whether eliminating bias and thus improving accuracy had any effect on the overall results. The Weibel-Gomez method estimates cell number from a single section through the glomerulus, whereas the disector/fractionator requires the glomerulus to be serially sectioned. Results. There was no significant difference between mean values obtained by the two methods, providing that the Weibel-Gomez estimate was performed on electron micrographs. However, the overall variance was high for all groups of patients, independent of the method employed. Conclusion. Although the disector/fractionator is the theoretic gold standard method for podocyte number estimation, comparable estimates can be obtained by the Weibel-Gomez method provided they are made from electron micrographs. Thus the technical resources available may determine the choice of method employed. Investigators should be aware of the high degree of variability in the estimate, particularly when trying to detect small changes in podocyte number.