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JHC exPRESS: First Published November 11, 2008. doi:10.1369/jhc.2008.952911
Journal of Histochemistry and Cytochemistry
Copyright © 2008 Burns et al.

A more recent version of this article appeared on March 1, 2009.
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Articles

Choice of Fixative Is Crucial to Successful Immunohistochemical Detection of Phosphoproteins in Paraffin-embedded Tumor Tissues

Janine A. Burns 1, Yuan Li 1, Carol A. Cheney 1, Yangsi Ou 1, Laura L. Franlin-Pfeifer 1, Nelly Kuklin 1 and Zhi-Qiang Zhang 1*

1 Department of Biologics Research, Merck Research Laboratories, West Point, Pennsylvania

* To whom correspondence should be addressed. E-mail: zhiqiang_zhang{at}merck.com.

Submitted on September 25, 2008
Accepted on 30 October 2008


  Abstract
Protein phosphorylation is frequently used as an indicator of cellular signaling activity. Elevated phosphorylation of tyrosine kinase receptors plays an important role in cancer pathogenesis. However, phosphoproteins are usually poorly preserved in clinical tissue samples that are routinely fixed in 10% formalin. Nonetheless, in oncology clinical trials, usage of phosphoproteins as biomarkers has been considered to be of great value in evaluating the effectiveness of a given drug candidate. Therefore, it is worthy of investigating whether alternative fixatives would improve the preservation of phosphoproteins in tissue. We compared the immunohistochemical staining of a number of phosphoproteins in xenograft and human surgical tumor tissues fixed in three different fixatives; 10% formalin, 4% paraformaldehyde (PFA), and Streck's tissue fixative (STF). We found that STF significantly enhanced the staining intensity of phosphoproteins as compared to 10% formalin or 4% PFA. STF fixative also demonstrated superiority of preservation of phosphoproteins in human surgical samples. Our results indicate that the choice of fixative could significantly affect the usability of clinical tissue samples for evaluating phosphoprotein by immunohistochemistry.

Key Words: fixative, STF, phosphoprotein, tumor tissues, paraffin tissue section


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J. Bodo, L. Durkin, and E. D. Hsi
Quantitative In Situ Detection of Phosphoproteins in Fixed Tissues Using Quantum Dot Technology
J. Histochem. Cytochem., July 1, 2009; 57(7): 701 - 708.
[Abstract] [Full Text] [PDF]



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