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JHC exPRESS: First Published June 13, 2005. doi:10.1369/jhc.5B6691.2005
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A more recent version of this article appeared on September 1, 2005.
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Brief Reports

Protein-embedding Technique: a Potential Approach to Standardization of Immunohistochemistry for Formalin-fixed, Paraffin-embedded Tissue Sections

Shan-Rong Shi 1, Cheng Liu 1, Jeanette Perez 1 and Clive R. Taylor 1*

1 Department of Pathology, University of Southern California, Keck School of Medicine, Los Angeles, California

* To whom correspondence should be addressed. E-mail: taylor{at}pathfinder.hsc.usc.edu.

Submitted on March 17, 2005
Accepted on 23 March 2005


  Abstract
A serial study was carried out to develop a protein-embedding technique for standardization of immunohistochemistry (IHC) on formalin-fixed, paraffin-embedded (FFPE) tissue sections. The primary goal was to find an optimal matrix material as a protein-carrier. This matrix must have two phases, a liquid phase to allow uniform mixing of a protein, and a solid phase forming a ‘block’ that can be fixed and processed in the same manner as human tissue. This standardized protein block would serve as a source of thin sections for control of IHC, and therefore must also withstand the boiling conditions of antigen retrieval (AR). After multiple experiments, a method was developed utilizing polymer microsphere (beads) as a support medium for protein. The resulting suspension of protein coated beads can then be condensed, fixed, embedded in paraffin, sectioned and applied as the standard reference material for routine IHC of FFPE tissue. Several proteins have been coated successfully onto the surface of the beads and have then been visualized as positive surface markers upon microscopy. The method showed particular promise for quantitative IHC of Her-2/neu, currently a major issue in clinical and research of breast cancer. The utility and limitations of AR can also be assessed by this method.

Key Words: immunohistochemistry, antigen retrieval, standardization, protein-embedding technique beads


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