The most complete, up-to-date reference on antigen retrieval andimmunohistochemistryAn antigen is a substance that prompts the generation ofantibodies and can cause an immune response. The antigen retrieval(AR) technique is in wide use across the globe, and is a criticaltechnique used in medical diagnosis of disease, particularlyclinical targeted cancer treatment.Antigen Retrieval Immunohistochemistry Based Research andDiagnostics discusses several scientific approaches to thestandardization of quantifiable immunohistochemistry (IHC). Basedon the development and application of AR by the editors, thisvolume summarizes recent achievements in AR-IHC and analyzesnumerous cutting-edge issues for future research projects.Featuring contributions from a worldwide group of leadingexperts and research scientists in the field, this importantwork:* Summarizes the key problems in the four fields of antigenretrieval* Discusses the advances of AR techniques and theirapplications* Provides practical methods and protocols in AR-IHC, such asextraction of nucleic acids and proteins for molecular analysis,cell/tissue sample preparation, and standardization and developmentof various techniques to meet the future needs of¿clinical andresearch molecular analysis* Encourages further research in AR and IHC, particularly how ARmethods might be employed for improved test performance and thedevelopment of greater reliability and reproducibility of IHC* Includes an appendix of related laboratory protocolsAntigen Retrieval Immunohistochemistry Based Research andDiagnostics is intended for clinical pathologists, molecularcell biologists, basic research scientists, technicians, andgraduate students who undertake tissue/cell morphologic andmolecular analysis and wish to use and extend the power ofimmunohistochemistry. It is also pertinent for most biotechnologycompanies majoring in development of IHC products.Wiley Series in Biomedical Engineering and Multi-DisciplinaryIntegrated Systems / Kai Chang, Series Editor

Preface ixContributors xvPart I Recent Advances in Antigen Retrieval Techniques and Its Application 11 Standardization of Antigen Retrieval Techniques Based on the Test Battery Approach 3Shan-Rong Shi and Clive R. Taylor2 Extended Application of Antigen Retrieval Technique in Immunohistochemistry and in SituHybridization 25Shan-Rong Shi and Clive R. Taylor3 Extraction of Dna/rna From Formalin-fixed, Paraffin-embedded Tissue Based on the Antigen Retrieval Principle 47Shan-Rong Shi and Clive R. TaylorPart II Standardization of Immunohistochemistry 734 Key Issues and Strategies of Standardization For Quantifiable Immunohistochemistry 75Shan-Rong Shi, Kevin A. Roth, and Clive R. Taylor5 Standardization of Immunohistochemistry Based on Antigen Retrieval Technique 87Shan-Rong Shi and Clive R. Taylor6 Standard Reference Material: Cell Line Development and Use of Reference Cell Lines as Standards for External Quality Assurance of Her2 Ihc and Ish Testing 101Bharat Jasani, Vicky Reid, Colin Tristram, Jeremy Walker, Paul Scorer, Michael Morgan, John Bartlett, Merdol Ibrahim, and Keith Miller7 Peptides as Immunohistochemistry Controls 123Steven A. Bogen and Seshi R. Sompuram8 Standard Reference Material: Protein- Embedding Technique and Design of Bar Code 141Shan-Rong Shi, Jiang Gu, and Clive R. Taylor9 the Pros and Cons of Automation for Immunohistochemistry From the Prospective Of the Pathology Laboratory 151David G. Hicks and Loralee McMahon10 Image Analysis in Immunohistochemistry 165Alton D. FloydPart III Tissue/cell Sample Preparation 18711 Tissue Cell Sample Preparation: Lessons From the Antigen Retrieval Technique 189Shan-Rong Shi and Clive R. Taylor12 Mechanisms of Action and Proper Use of Common Fixatives 195Richard W. Dapson13 Cell Sample Preparation for Clinical Cytopathology: Current Status and Future Development 219Yan Shi and Patricia G. Wasserman14 Design of a Tissue Surrogate to Examine Accuracy of Proteomic Analysis 235Carol B. Fowler, Jeffrey T. Mason, and Timothy J. O'learyPart IV Molecular Mechanism of Antigen Retrieval Technique 25115 Study of Formalin Fixation and Heat-induced Antigen Retrieval 253Jeffrey T. Mason, Carol B. Fowler, and Timothy J. O'leary16 a Linear Epitopes Model of Antigen Retrieval 287Steven A. Bogen and Seshi R. Sompuram17 Ph Or Ionic Strength of Antigen Retrieval Solution: a Potential Role for Refolding During Heat Treatment 303Shuji Yamashita18 Commentary: Future Directions 323Timothy J. O'Leary, Carol B. Fowler, David L. Evers, Robert E. Cunningham, and Jeffrey T. MasonPart V Proteomic Analysis of Protein Extracted From Tissue/cells 33319 Techniques of Protein Extraction From Ffpe Tissue/cells for Mass Spectrometry 335Carol B. Fowler, Timothy J. O'Leary, and Jeffrey T. Mason20 Application of Shotgun Proteomics to Formalin-fixed and Paraffin-embedded Tissues 347Brian M. Balgley21 Visualizing Protein Maps in Tissue 369Masahiro Mukai and Mitsutoshi Setou22 Symbiosis of Immunohistochemistry and Proteomics: Marching to a New Era 391Shan-Rong Shi, Brian M. Balgley, and Clive R. TaylorAppendix Related Laboratory Protocols 399Index 413