title: Design-based stereological methods for estimating numbers of glomerular podocytes.
abstract: 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.
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia. Electronic address: victor.puellesrodriguez@monash.edu.
authors.forename: Victor G
authors.initials: VG
authors.lastname: Puelles
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia.
authors.forename: Rebecca N
authors.initials: RN
authors.lastname: Douglas-Denton
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia.
authors.forename: Luise
authors.initials: L
authors.lastname: Cullen-McEwen
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia.
authors.forename: Bridgette J
authors.initials: BJ
authors.lastname: McNamara
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia.
authors.forename: Firuz
authors.initials: F
authors.lastname: Salih
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Pathology, University of Mississippi Medical Center, Jackson, MS, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Hughson
authors.affiliation: Centre for Chronic Disease, The University of Queensland, Brisbane, Australia.
authors.forename: Wendy E
authors.initials: WE
authors.lastname: Hoy
authors.affiliation: Stereology and EM Laboratory, CSGB, Aarhus University Hospital, Aarhus, Denmark.
authors.forename: Jens R
authors.initials: JR
authors.lastname: Nyengaard
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia.
authors.forename: John F
authors.initials: JF
authors.lastname: Bertram
mesh_terms: D000818:Animals; D001706:Biopsy; D002452:Cell Count; D006801:Humans; D007150:Immunohistochemistry; D007678:Kidney Glomerulus / Q000166:cytology*; D008853:Microscopy; D018613:Microscopy, Confocal; D050199:Podocytes / Q000502:physiology*
keywords: Glomerular volume; Immunohistochemistry and confocal microscopy; Podocyte; Stereology