Handbook of Practical Immunohistochemistry: Frequently Asked Questions

756 Pages·2015·67.29 MB·English

by  Fan Lin

Fan Lin
Jeff rey Prichard
Editors
Haiyan Liu
Myra Wilkerson
Associate Editors
Zongming E. Chen
Handbook of Practical
Immunohistochemistry
Frequently Asked Questions
Second Edition
123
Handbook of Practical Immunohistochemistry

Fan Lin • Jeffrey Prichard
Editors
Handbook of Practical
Immunohistochemistry
Frequently Asked Questions
Second Edition
Editors
Fan Lin Jeffrey Prichard
Laboratory Medicine Laboratory Medicine
Geisinger Health System Geisinger Medical Center
Danville , PA , USA Danville , PA , USA
ISBN 978-1-4939-1577-4 ISBN 978-1-4939-1578-1 (eBook)
DOI 10.1007/978-1-4939-1578-1
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2015933831
© Springer Science+Business Media New York 2015
T his work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is
concerned, specifi cally the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction
on microfi lms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation,
computer software, or by similar or dissimilar methodology now known or hereafter developed. Exempted from this
legal reservation are brief excerpts in connection with reviews or scholarly analysis or material supplied specifi cally
for the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work.
Duplication of this publication or parts thereof is permitted only under the provisions of the Copyright Law of the
Publisher’s location, in its current version, and permission for use must always be obtained from Springer. Permissions
for use may be obtained through RightsLink at the Copyright Clearance Center. Violations are liable to prosecution
under the respective Copyright Law.
T he use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not
imply, even in the absence of a specifi c statement, that such names are exempt from the relevant protective laws and
regulations and therefore free for general use.
While the advice and information in this book are believed to be true and accurate at the date of publication, neither
the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may
be made. The publisher makes no warranty, express or implied, with respect to the material contained herein.
Printed on acid-free paper
Springer is part of Springer Science+Business Media (www.springer.com)
Pref ace
As in the fi rst edition, the second edition of H andbook of Practical Immunohistochemistry—
Frequently Asked Questions is written in a question and answer (Q&A) format and intended to
be a practical, user-friendly, quick reference for information related to using the most up-to-
date immunohistochemistry in clinical diagnosis. The new edition demonstrates a signifi cant
revision and improvement over the fi rst edition in many ways summarized as follows:
1. More Chapters : Four new chapters have been added: (1) Geisinger IHC Antibodies and
Staining Protocols; (2) “Rabbit Monoclonal Antibodies”; (3) “Immunohistochemical
Detection of Phosphoproteins and Cancer Pathways”; and (4) “Overview of Predictive
Biomarkers and Integration of Immunohistochemistry into Molecular Pathology.”
Extensive additions and changes have been made to the Chap. 2 , and the title of the chapter
was changed to “Standardization of Diagnostic Immunohistochemistry.”
2. More questions and answers : Over 100 questions and answers have been added to the new
edition. Nearly all chapters have been updated to include relevant new questions, new
markers, more refi ned IHC panels, representative pictures, and current references. For
instance, in the “Unknown Primary” chapter, 34 additional questions, 3 new diagnostic
algorithms, 10 color pictures, and 150 new references have been added to the chapter.
3. More refi ned working antibody panels: When you examine each individual table, you will
notice that some of the antibodies in the table are highlighted. The highlighted set of anti-
bodies is the suggested and refi ned panel for an initial workup. Brief notes are provided
for many tables in order to reiterate the most important diagnostic applications and pitfalls
that one may encounter.
4. More new diagnostic and predictive biomarkers: Many recently described antibodies in
clinical IHC laboratories have been added. These include both diagnostic markers and
predictive biomarkers such as GATA3, NY-BR-1, PAX8, PAX2, napsin A, p40, glucose
transporter 1, CD146, ERG, NKX3.1, NKX2.2, DOG1, SATB2, cadherin 17, arginase-1,
pVHL, maspin, S100P, UP II, IMP3, SALL4, OCT4, LIN28, Nanog, SF-1, SOX2, HNF-
1beta, FOXL2, HGAL, LMO2, LEF1, CD160, SOX11, IRTA1, TROP2, adipophilin,
SOX10, Merkel cell polyomavirus, MUC4, NY-ESO-1, INI-1, STAT6, SDHB, BRAF,
EGFR, ALK, c-MYC, etc.
5. More high-quality color pictures from Geisinger Medical Laboratories (GML) IHC
slides: An extensive set of high-quality color pictures and diagnostic algorithms, if avail-
able, is included in each chapter to illustrate some of the key antibodies, including many
recently discovered and substantiated antibodies used in that particular chapter. There are
over 700 color pictures, representing a signifi cant increase over the fi rst edition, taken
from GML IHC slides using the recommended staining protocols contained in the appen-
dices of this book.
6. More GML data : In many tables you will see comparisons of data from GML to data from
the literature. This is a unique feature of this book. The reproducibility of antibodies
reported in the literature is sometimes in question; to improve the reproducibility, we have
undertaken the daunting task of testing the antibodies listed in the appendices using more
v
vi Preface
than 7,000 TMA slides and 1,500 routine slides. These TMA sections contain thousands
of tumors from various organs and normal tissues in the GML archives. If your lab follows
the protocols in the appendices, you should obtain results similar to ours.
7. IHC on normal tissues : Immunophenotypes of many normal tissues, which receive little
or no attention in other surgical pathology and IHC books, have been included in many
chapters, such as normal breast, lung, pancreas, ampulla, colon, stomach, small intestine,
kidney, etc.
8. More current references : A large number of current references have been included in each
new and updated chapter.
9. Better Index : A signifi cant amount of time and effort has been devoted to creating a better
and more useful index. The index in the second edition encompasses an extensive list of
common and rare diagnostic entities; frequently used, rare, and newly described antibod-
ies; and many keywords which a practicing pathologist may use to search for a specifi c
question.
10. Updated IHCFAQ.com website : An updated free companion IHC website (IHCFAQ.com)
contains additional stain images and staining protocols for many new antibodies.
11. Data interpretation : To standardize our manual scoring system, unless otherwise speci-
fi ed, the following criteria are applied throughout this book:
(a) − = usually less than 5 % of cases are stained;
(b) + = usually greater than 70 % of cases are stained;
(c) + or − = usually more than 50 % but less than 70 % of cases are stained;
(d) − or + = usually less than 50 % of cases are stained;
(e) V = variable, or sometimes positive; data are somewhat inconsistent;
(f) ND = no data available.
12. More expert contributions : Last but not least, many chapters also include contributions
from an expert in his or her fi eld.
A fter publication of the fi rst edition, we received overwhelmingly positive feedback and
valuable suggestions and comments from readers, and we would like to take this opportunity
to express our very sincere thanks to all of you. As always, the editors ask for your understanding
of any potential errors in this new edition and also invite you to submit your feedback,
suggestions, and comments to us ([email protected]; [email protected]; Hliu1@
geisinger.edu; [email protected]; [email protected]). With continued support
from readers like you, we are confi dent that each future edition will be even more complete and
informative than the previous one.
Danville, PA Fan Lin, MD, PhD
Jeffrey Prichard, DO
Haiyan Liu, MD
Myra L. Wilkerson, MD
Zongming E. Chen, MD, PhD
Acknowledgments
Producing the second edition of this IHC Handbook was as challenging as publishing the fi rst
edition for our department and we wish to acknowledge the assistance and tremendous support
we received from our staff. Therese Snyder, Vice President of Laboratory Operations, supported
and encouraged this project from conception through completion. Sandy Mullay, Operations
Director of Anatomic Pathology, always ensured we had the technical, secretarial, and clerical
support needed for all phases of the project. Melissa Erb served as our project coordinator,
collecting hundreds of new references, scheduling numerous meetings, keeping us organized,
and moving the project forward. Kathy Fenstermacher was invaluable in editing, formatting,
and polishing book chapters, as well as producing many diagrams. Without Melissa and
Kathy’s tireless effort, the second edition would not be possible to meet the submission
deadline. Erin Powell and John Shaw constructed many TMA blocks for testing and studying
many new antibodies. Angela Bitting and Jianhui Shi are responsible for bringing you the
hundreds of optimized IHC protocols available in the appendices of this book. Tina Brosious,
Glen Kauwell, Laurie Kneller-Walter, Deanna Ward, Beth Reigle, Kris Bricker, and Kathryn
Walker, immunotechnologists helped stain numerous tissue microarray sections and routine
slides. Mary Sejuit spent countless hours pulling and refi ling slides and paraffi n blocks,
keeping everything well organized. Christy Attinger provided expert secretarial support.
Kristen Stiver, Supervisor of Anatomic Pathology, helped with scheduling and coordination of
technical staff. Finally, we are in debt to our families and close friends for their understanding
that we have again buried ourselves in this project for such a long period of time. We are very
fortunate to have your love and incredible support.
Fan Lin, MD, PhD
Jeffrey Prichard, DO
Haiyan Liu, MD
Myra L. Wilkerson, MD
Zongming E. Chen, MD, PhD
vii

Contents
1 Immunohistochemistry Quality Management and Regulation ........................... 1
Jeffrey Prichard
2 Standardization of Diagnostic Immunohistochemistry ........................................ 17
Fan Lin and Jianhui Shi
3 Automated Immunohistochemistry Overview ...................................................... 31
Jeffrey Prichard and Angela K. Bitting
4 Geisinger Immunohistochemical Antibodies and Staining Protocols ................. 39
Fan Lin , Jeffrey Prichard , Angela K. Bitting , Jianhui Shi , Haiyan Liu ,
Myra L. Wilkerson , and Zongming E. Chen
5 Immunohistochemistry: A Dako Perspective ........................................................ 57
Ole Feldballe Rasmussen and Lars Rudbeck
6 Immunohistochemistry: Ventana Perspective ....................................................... 69
Traci DeGeer
7 Rabbit Monoclonal Antibodies ............................................................................... 77
Weimin Zhu , Aihua Li , and Taiying Chen
8 Immunohistochemical Detection of Phosphoproteins
and Cancer Pathways .............................................................................................. 85
Kerstin A. David and Hartmut Juhl
9 Overview of Predictive Biomarkers and Integration
of IHC into Molecular Pathology ........................................................................... 91
Zongming E. Chen and Fan Lin
10 Tissue Microarray .................................................................................................... 105
Myra L. Wilkerson and Stephen M. Hewitt
11 Unknown Primary/Undifferentiated Neoplasm .................................................... 119
Fan Lin and Haiyan Liu
12 Exfoliative Cytology and Effusions ........................................................................ 165
Steven C. Meschter and Jan F. Silverman
13 Breast......................................................................................................................... 1 83
Haiyan Liu
ix