Original ContributionsInadequate formalin fixation decreases reliability of p27Kip1 immunohistochemical staining: Probing optimal fixation time using high-density tissue microarrays*,**
Section snippets
Specimen handling and TMA construction
In our standard approach to radical prostatectomy sectioning, the gland is inked, and the specimens are fixed for 1 to 4 hours in formalin by simply placing the specimen in a minimum of 15 volumes of 10% neutral buffered formalin. Prostate specimens are then sectioned from apex to base into 3- to 5-mm slices, and each slice is further sectioned into halfs or, more commonly quadrants. Tissues are submitted into standard-sized cassettes, and are placed onto our automated tissue processors the
Results
The gradation of staining seen with the standard sections from a radical prostatectomy sample sectioned and cut using our routine protocol of same-day tissue processing is illustrated in Fig 1.
Discussion
In this study, we show that immunohistochemical staining against p27Kip1 protein using radical prostatectomy specimens in formalin-fixed tissue may mistakenly appear to be downregulated as a result of an artifact relating to inadequate tissue fixation. Contrary to popular dogma in the pathology field, prolonged fixation, even up to 8 days, produced no decrement in staining for p27Kip1. Although TMAs are not necessary to carry out studies on the effects of fixatives on immunostaining, they do
Acknowledgements
The authors thank Geert van Leenders for illustrating, first-hand, the method of formalin injection of radical prostatectomy specimens.
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Supported by grants from Public Health Services NIH/NCI Specialized Program in Research Excellence (SPORE) in Prostate Cancer, Johns Hopkins (P50CA58236), and The University of Michigan (P50CA69568).
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Address correspondence and reprint requests to Angelo M. De Marzo, MD, PhD, Johns Hopkins Medical Institutions, Department of Pathology, Division of Genitourinary Pathology, Bunting/Blaustein Cancer Research Building, Room 153, 1650 Orleans St, Baltimore, MD 21231-1000.