Design-based stereological methods for estimating numbers of glomerular podocytes

Victor G. Puelles, Rebecca N. Douglas-Denton, Luise Cullen-McEwen, Bridgette J. McNamara, Firuz Salih, Jinhua Li, Michael D. Hughson, Wendy E. Hoy, Jens R. Nyengaard, John F. Bertram

The podocyte depletion hypothesis has emerged as a unifying concept in glomerular pathology. According to this hypothesis podocyte depletion may be absolute (decrease in number of healthy mature podocytes), relative (fewer podocytes per unit of glomerular volume) or involve alterations to the specialized podocyte architecture (such as foot process effacement). To study and understand podocyte depletion it is important to be able to accurately and precisely count these cells. Here we present new design-based stereological methods for estimating podocyte number in individual glomeruli of known volume, and in average glomeruli. Both methods involve serial histological sectioning, triple label immunohistochemistry, laser confocal microscopy and cell counting with the optical disector/fractionator.