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JHC exPRESS: First Published September 7, 2005. doi:10.1369/jhc.5A6758.2005
Copyright © Histochemical Society, Inc.

A more recent version of this article appeared on January 1, 2006.
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Computerized Morphometric Analysis of Pathologic Prion Protein Deposition in Scrapie Infected Hamster Brain

Olga A. Maximova 1*, Rolf E. Taffs 1, Kitty L. Pomeroy 1, Pedro Piccardo 1 and David M. Asher 1

1 Laboratory of Bacterial, Parasitic and Unconventional Agents, Division of Emerging and Transfusion-Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, United States Food and Drug Administration, Rockville, Maryland

* To whom correspondence should be addressed. E-mail: maximova{at}cber.fda.gov.

Submitted on June 14, 2005
Accepted on 23 August 2005


  Abstract
Transmissible spongiform encephalopathies (TSEs or prion diseases) are characterized by a constellation of typical though variable pathological changes in the brain. Deposition of disease-associated abnormal prion protein (PrPSc) is the pathological feature of TSEs most consistent and accessible for quantification. However, the evaluation of PrPSc deposits detected by immunohistochemical techniques has been traditionally based on arbitrarily assigned semi-quantitative scores. This approach is limited by its subjectivity and bias, yielding considerable variability. In this study, we used MetaMorph 6.1 image analysis software for quantitative analysis of immunostained PrPSc deposits in the CNS of hamsters infected with the 263K strain of scrapie agent. Computerized morphometric analysis (CMA) allowed unambiguous detection of even minimal amounts of immunostained PrPSc. CMA values for intensity of staining and area stained correlated well with semi-quantitative scores, providing reproducible quantitative data and objective criteria for analyzing PrPSc deposition. CMA provides a simple and reliable method for improved and consistent diagnosis of TSEs that may also be used to quantify other immunostained biomarkers.

Key Words: digital imaging, hue-saturation-intensity, immunohistochemistry, morphometry, prion disease, scrapie prion protein, transmissible spongiform encephalopathy


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