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2015v1.0
Bancroft’s
THEORY and
PRACTICE
of HISTOLOGICAL
TECHNIQUES
This page intentionally left blank
Bancroft’s
THEORY and
PRACTICE
of HISTOLOGICAL
TECHNIQUES
EIGHTH
EDITION
S. Kim Suvarna
Consultant Pathologist,
Histopathology Department,
Sheffield Teaching Hospitals
Sheffield, UK
Christopher Layton
Specialist Section Lead in Specimen Dissection,
Histopathology Department,
Sheffield Teaching Hospitals
Sheffield, UK
John D. Bancroft
Retired Pathology Directorate Manager and Business Manager,
Queen’s Medical Centre,
Nottingham, UK
© 2019, Elsevier Limited. All rights reserved.
First edition 1977
Second edition 1982
Third edition 1990
Fourth edition 1996
Fifth edition 2002
Sixth edition 2008
Seventh edition 2013
The rights of Dr. S. Kim Suvarna, Dr. Christopher Layton and Mr. John D. Bancroft to be identified as
authors of this work has been asserted by them in accordance with the Copyright, Designs and Patents
Act 1988.
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or mechanical, including photocopying, recording, or any information storage and retrieval system,
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This book and the individual contributions contained in it are protected under copyright by the
Publisher (other than as may be noted herein).
Notices
Knowledge and best practice in this field are constantly changing. As new research and experience
broaden our understanding, changes in research methods, professional practices, or medical treat-
ment may become necessary.
Practitioners and researchers must always rely on their own experience and knowledge in evalu-
ating and using any information, methods, compounds, or experiments described herein. In using
such information or methods they should be mindful of their own safety and the safety of others,
including parties for whom they have a professional responsibility.
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Preface to the eighth edition
It is now forty years since the first edition of this simply reviewed these to ensure that their modern
book was published, and the histological labora- relevance has been achieved. The previous edi-
tory has changed dramatically in that time. Whilst tion’s three chapters on immunohistochemistry,
some techniques of tissue selection, fixation and immunofluorescence and quality control have been
section production have remained reassuringly amalgamated into one chapter and digital pathol-
constant, there have been great advances in terms ogy replaces the quantitative data from microscopic
of immunological, molecular diagnostic and digital specimens. Microarray is now an appendix.
methodology. Immunohistochemistry and immu- There are several new contributors for this edi-
nofluorescence now have well-defined diagnostic tion. They include updates on the management
and screening roles with quality assurance reali- chapter by Beth Cox and Emma Colgen, and labo-
ties, and are to be found throughout the world with ratory safety by Ada Feldman. The fixation chapter
pivotal interactions in patient management. In par- has been updated by the editors. The immunohisto-
ticular, the progressive development of molecular chemistry and immunofluorescent chapter has been
techniques over the last 20 years, revolving around updated by the previous authors along with Ann
DNA and in situ hybridization has permitted the Michelle Cull and Jennifer Marston.
creation of new genetic tests and diagnostic oppor- The new chapter on automation is written by Greg
tunities for the laboratory. These are currently at the Zardin and Lynne Braithwaite, and digital pathol-
forefront of guiding treatment choices for patients. ogy by Jonathan Bury and Jonathan Griffin. Phillipe
At the same time, this has dictated the rational Taniere, Brendan O’Sullivan, Matthew Evans and
review of some classic histological tests resulting Frances Hughes have rewritten and updated the
in a reduced histochemical repertoire which is the molecular pathology chapter.
reality in many laboratories. Digital pathology in Having said this, we are conscious that we are all
particular is the new frontier much as PACS was part of the lineage of authors who have contributed
to radiology 10 years ago. It is likely that the next to the previous editions of this book. We salute and
edition will have a consolidated approach to this thank them for their work. Indeed, their contribu-
exciting new technique. tion to the success of this ongoing text cannot be
As always, acknowledgment of the old as well as underestimated.
the new diagnostic methodology will be required by Ultimately, we hope that we have produced a
both trained and trainee staff within the histopathol- modern and relevant histotechnology text which
ogy laboratory and scientists in related fields. will be of use to those in training as well as estab-
As in the 7th edition the classical and now rarely lished practitioners worldwide. As always, we rec-
used staining methods are in the appendices but ognize that this edition is but one step of the ongoing
where the reader needs more information, a refer- story and hope that our international colleagues will
ence to earlier editions is made. This has allowed for enjoy and approve of the changes which have taken
further expansion and update in the newer diagnos- place.
tic methodologies. S. Kim Suvarna, Christopher Layton and
We recognized that some sections on classical John D. Bancroft
stains have not changed dramatically and have March 2018
v
Preface to the first edition
In recent years histological techniques have become elsewhere. Although the content is particularly
increasingly sophisticated, incorporating a whole suitable for students working towards the Special
variety of specialties, and there has been a corre- Examination in Histopathology of the Institute of
sponding dramatic rise in the level and breadth of Medical Laboratory Sciences, the level is such that
knowledge demanded by the examiner of trainees in more advanced students, along with research work-
histology and histopathology technology. ers, histologists, and pathologists, will find the book
We believe that the time has arrived when no beneficial. To achieve this we have gathered a team
single author can produce a comprehensive book of expert contributors, many of whom have writ-
on histology technique sufficiently authoritative in ten specialized books or articles on their own sub-
the many differing fields of knowledge with which ject; most are intimately involved in the teaching of
the technologist must be familiar. Many books exist histology and some are examiners in the HNC and
which are solely devoted to one particular facet such Special Examination in Histopathology. The medi-
as electron microscopy or autoradiography, and the cally qualified contributors are also involved in tech-
dedicated technologist will, of course, read these in nician education.
the process of self-education. Nevertheless the need All contributors have taken care to give, where
has arisen for a book which covers the entire spec- applicable, the theoretical basis of the techniques,
trum of histology technology, from the principles of for we believe that the standard of their educa-
tissue fixation and the production of paraffin sec- tion has risen so remarkably in recent years that
tions to the more esoteric level of the principles of the time is surely coming when medical laboratory
scanning electron microscopy. It has been our aim technicians will be renamed ‘medical laboratory sci-
then, to produce a book which the trainee technolo- entists’; we hope that the increase in ‘scientific’ con-
gist can purchase at the beginning of his career and tent in parts of this book will assist in this essential
which will remain valuable to him as he rises on the transformation.
ladder of experience and seniority. John D. Bancroft
The book has been designed as a comprehen- Alan Stevens
sive reference work for those preparing for exami- Nottingham, 1977
nations in histopathology, both in Britain and
vi
List of contributors
The editor(s) would like to ac- Matthew Evans MA MBBS Frances Hughes CSci FIBMS
knowledge and offer grateful KS AKC Senior Biomedical Scientist in
thanks for the input of all previ- Molecular Pathology Diagnostic Molecular Pathology
ous editions’ contributors, without Service Molecular Pathology Diagnostic
whom this new edition would not Cellular Pathology Service
have been possible. Queen Elizabeth Hospital University Hospitals Birmingham
Birmingham, UK Birmingham, UK
John D. Bancroft
Retired Pathology Directorate Ada T. Feldman MS Stuart Inglut BSc (Hons)
Manager and Business Manager HT/HTL(ASCP) Histopathology Department
Queen’s Medical Centre CEO Glangwili General Hospital
Nottingham, UK Anatech Ltd Carmarthen
Battle Creek Wales, UK
Lynne Braithwaite BSc MSc MI, USA
CSci FIBMS Gayti B. Morris BA MBBCh
Advanced Practitioner Janet A. Gilbertson CSci FRCPath
Histopathology FIBMS Consultant Microbiologist
Pathlinks Principal Scientist Microbiology Department
Lincoln, UK National Amyloidosis Centre Sheffield Teaching Hospitals
University College London Sheffield, UK
Jonathan Bury BMedSci Royal Free
MBChB MPhil FRCPath London, UK Christopher Layton PhD
Consultant Histopathologist Specialist Section Lead in
Department of Pathology Jonathan Griffin MBChB Specimen Dissection
Sheffield Teaching Hospitals; (Hons) Histopathology Department
Honorary Senior Lecturer Specialty Registrar in Sheffield Teaching Hospitals
Department of Oncology Histopathology Sheffield, UK
University of Sheffield Sheffield Teaching Hospitals
Sheffield, UK Sheffield, UK Jennifer Marston MIBMS
BSc MSc
Emma Colgan MBA MSc J. Robin Highley DPhil Specialist Biomedical Scientist
FIBMS FRCPath Histopathology Department
Directorate Manager and Senior Clinical Lecturer in Sheffield Teaching Hospitals
Professional Lead Neuropathology Sheffield, UK
Laboratory Medicine The Sheffield Institute for
Sheffield Teaching Hospitals Translational Neuroscience Danielle McCluskey Bsc Msc
Sheffield, UK Sheffield, UK MIBMS
Advanced Biomedical Scientist
Beth Cox BS HTL/SCT(ASCP) Richard W. Horobin BSc PhD Histopathology
QIHC Honorary Research Fellow Central Manchester University
AP Consultant Chemical Biology and Medicinal Hospitals NHS Foundation Trust
Pathology Solutions Inc. Chemistry Manchester, UK
West Branch School of Chemistry
MI, USA University of Glasgow Ann Michelle Cull BSc
Glasgow, UK (Hons) MSc
Histopathology Department
Sheffield Teaching Hospitals
Sheffield, UK
vii
vviiiiii List of contributors
Guy E. Orchard PhD MSc Sophie R. Stenton MBChB Philippe Taniere MD PHD
(dist) FIBMS BMedSci Molecular Pathology Diagnostic
Consultant Grade Biomedical Department of Paediatric Service
Scientist and Laboratory Manager Pathology Cellular Pathology
Tissue Sciences Viapath; Sheffield Children’s Hospital Queen Elizabeth Hospital
Histopathology, St. John’s Institute Sheffield, UK Birmingham, UK
of Dermatology
St. Thomas’ Hospital Diane L. Sterchi MS Eu-Wing Toh MBBS BMedSci
London, UK HTL(ASCP) MD
Senior Research Associate Histopathology Department
Brendan O’Sullivan BSc Histomorphometry Lead Sheffield Teaching Hospitals
(Hons) Department of Pathology Sheffield, UK
Operations Manager Covance Laboratories Inc.
Molecular Pathology Greenfield Graeme Wild BSc PhD
Cellular Pathology IN, USA Immunology Department
University Hospitals Birmingham Sheffield Teaching Hospitals
NHS Foundation Trust John W. Stirling BSc (Hons) Sheffield, UK
Birmingham, UK M.Lett
Honorary Lecturer Dee Wolfe AS HT(ASCP)QIHC
Elisabeth J. Ridgway MBBS Molecular Medicine and Pathology Vice President
BSc MD FRCPath Flinders University Technical Service
Consultant Microbiologist Adelaide, SA, Australia Anatech Ltd
Microbiology Department Battle Creek
Sheffield Teaching Hospitals Jennifer H. Stonard MSc MI, USA
Sheffield, UK CSci MIBMS
Specialist Biomedical Scientist Anthony E. Woods BA
Paul Samuel BSc DMLT Cellular Pathology BSc(Hons) PhD MAIMS
MIBMS John Radcliffe Hospital; FFSc(RCPA)
Histopathology Department Specialist Biomedical Scientist Associate Professor
Sheffield Teaching Hospitals Histopathology Associate Head: School of
Sheffield, UK Nuffield Orthopaedic Centre Pharmacy and Medical Sciences
Oxford, UK University of South Australia
Tracy Sanderson FIBMS Adelaide, SA, Australia
IHC Scientific Lead Nicky Sullivan CSci FIBMS
Histopathology Department of Neuropathology Greg Zardin BSc (Hons) MSc
Sheffield Teaching Hospitals and Ocular Pathology Advanced BMS
Sheffield, UK John Radcliffe Hospital Histopathology
Oxford, UK Sheffield Teaching Hospitals
Lena T. Spencer MA Sheffield, UK
HTL(ASCP)QIHC S. Kim Suvarna MBBS BSc
Senior Histotechnologist FRCP FRCPath
Anatomic Pathology Consultant Pathologist
Norton Healthcare Histopathology Department
Louisville Sheffield Teaching Hospitals
KY, USA Sheffield, UK
