Hum. Reprod. Advance Access published online on June 4, 2007
Human Reproduction, doi:10.1093/humrep/dem137
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Estimating human ovarian non-growing follicle number: the application of modern stereology techniques to an old problem
1 Stereotome NW, Issaquah, WA 98027, USA 2 Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, University of Oklahoma Health Sciences Center, PO Box 26901, Oklahoma City, OK 73190, USA 3 Seattle Reproductive Medicine, Seattle, WA 98109, USA 4 Inserm U-407, 69921 Oullins, France 5 Children's Hospital and Regional Medical Center, Seattle, WA 98105, USA
6 Correspondence address. Tel: +1-405-271-8722; Fax: +1-405-271-1655; E-mail: karl-hansen{at}ouhsc.edu
BACKGROUND: Previous published reports on the number of non-growing follicles (NGFs) in the human ovary have employed model-based methods for number estimates. These methods are time-intensive, and require correction factors and assumptions that ultimately limit their accuracy. Here, we describe the modification, application and validation of a modern fractionator/optical disector technique for the estimation of human ovarian NGF number.
METHODS: Forty-eight pairs of normal human ovaries were collected from women (age 8–51 years) undergoing elective bilateral oophorectomy, organ donation, or from autopsy. After gross pathologic examination, systematic random sampling was utilized to obtain tissue for analysis by the fractionator/optical disector method. The precision of individual NGF counts was determined by calculating the observed coefficient of error (OCE). Intra-observer variability and variation in NGF number between ovaries within a pair were also determined.
RESULTS: The mean OCE was 16.6% with larger variations observed at lower follicle counts. In recount experiments of the same ovary, NGF number estimates varied by 15–29%, except at very low follicle counts where variation was greater, but absolute differences were small. There was no significant difference in NGF number between ovaries within a pair (Wilcoxon signed rank test, P = 0.81).
CONCLUSIONS: Modern stereology methods provide an unbiased, efficient method for estimating NGF number in the human ovary. Both ovaries within a pair contain similar numbers of NGFs.
Key words: optical disector/fractionator/follicles/human ovary/stereology
Presented in part at the 2003 Society for Gynecological Investigation annual meeting. Submitted on January 25, 2007; resubmitted on March 8, 2007; accepted on April 11, 2007.
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