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Toxicologic Pathology
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Articles

Ovarian Follicle Counts Using Proliferating Cell Nuclear Antigen (PCNA) and Semi-Automated Image Analysis in Rats

Catherine A. Picut1, Cynthia L. Swanson1, Kathryn L. Scully1, Vern C. Roseman2, Regina F. Parker1 and Amera K. Remick1

1 WIL Research Laboratories–Biotechnics, LLC, Hillsborough, North Carolina, USA
2 Media Cybernetics, Bethesda, Maryland, USA

Correspondence: Catherine A. Picut, WIL-Biotechnics, LLC, 310 Millstone Drive, Hillsborough, NC 27278; e-mail:cpicut{at}biotechnics-inc.com.

Ovarian follicle counting is a method to assess ovarian toxicity in reproductive toxicity studies in rats. Although ovarian follicle counting has been traditionally performed manually on hematoxylin and eosin (H&E)-stained sections, the use of immunohistochemical methods, including human cytochrome P450 1B1 (CYP1B1) and proliferating cell nuclear antigen (PCNA), have been used to enhance the visibility of the primordial and primary follicles to facilitate manual counting. In this study, serial sections from both ovaries from ten 3-month-old female Sprague Dawley rats were stained using routine H&E and immunohistochemistry for PCNA. Counting of primordial and primary follicles was performed manually using these two stains and by semi-automated image analysis of PCNA-stained slides. Although manual counting of PCNA-stained slides is preferable to manual counting of H&E-stained slides, manual counting involves variability between individual counters. Semi-automated image analysis of PCNA-stained slides yields an accurate and consistent count of these primordial/primary follicles and eliminates variability between individual counters.

Key Words: ovarian follicle • proliferating cell nuclear antigen (PCNA) • image analysis • immunohistochemistry • reproductive toxicology

Abbreviations: H&E, hematoxylin and eosin • PCNA, proliferating cell nuclear antigen

This version was published on July 1, 2008

Toxicologic Pathology, Vol. 36, No. 5, 674-679 (2008)
DOI: 10.1177/0192623308317428
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