Table Of ContentDiagnostic Electron
Microscopy – A Practical
Guide to Interpretation
and Technique
CurrentandfuturetitlesintheRoyalMicroscopicalSociety –
JohnWileySeries
Published
Principles and Practice of Variable Pressure/Environmental Scanning
ElectronMicroscopy(VP-ESEM)
DebbieStokes
Aberration-CorrectedAnalyticalElectronMicroscopy
EditedbyRikBrydson
Diagnostic Electron Microscopy – A Practical Guide to Interpretation
andTechnique
EditedbyJohnW.Stirling,AlanCurry&BrianEyden
Forthcoming
LowVoltageElectronMicroscopy:PrinciplesandApplications
EditedbyDavidC.Bell&NatashaErdman
AtlasofImagesandSpectraforElectronMicroscopists
EditedbyUrselBangert
UnderstandingPracticalLightMicroscopy
JeremySanderson
FocusedIonBeamInstrumentation:TechniquesandApplications
DudleyFinch&AlexanderBuxbaum
ElectronBeam-SpecimenInteractionsandApplicationsinMicroscopy
BudhikaMendis
Diagnostic Electron
Microscopy – A Practical
Guide to Interpretation
and Technique
Edited by
John W. Stirling
TheCentreforUltrastructuralPathology,Adelaide,Australia
Alan Curry
ManchesterRoyalInfirmary,Manchester,UK
and
Brian Eyden
ChristieNHSFoundationTrust,Manchester,UK
Published in association with the Royal
Microscopical Society
Series Editor: Susan Brooks
A John Wiley & Sons, Ltd., Publication
Thiseditionfirstpublished2013
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LibraryofCongressCataloging-in-PublicationData
Diagnosticelectronmicroscopy:apracticalguidetointerpretationandtechnique/editedby
JohnW.Stirling,AlanCurry,andBrianEyden.
p.;cm.
Includesbibliographicalreferencesandindex.
ISBN978-1-119-97399-7(cloth)
I.Stirling,JohnW.II.Curry,Alan.III.Eyden,Brian.
[DNLM:1.DiagnosticImaging--methods.2.Microscopy,Electron,Transmission.WN180]
(cid:1)
616.0754–dc23
2012027835
AcataloguerecordforthisbookisavailablefromtheBritishLibrary.
ISBN:978-1-119-97399-7
Setin10.5/13ptSabonbyLaserwordsPrivateLimited,Chennai,India
Acknowledgements
and Dedication
All three editors wish to thank the many individuals who have helped
to make this volume possible. Firstly, they would like to express their
appreciation to all the authors for their hard work and generosity in
sharing their professional experience, as well as all the ‘behind-the-
scenes’ staff and colleagues without whom this book could not have
beenproduced.
JohnStirlingthanksthestaffoftheCentreforUltrastructuralPathol-
ogy,SAPathology,Adelaide,fortheirsupportandphotographiccontri-
butions – especially Alvis Jaunzems and Jeffrey Swift – and DrSophia
Otto of the Department of Surgical Pathology, SA Pathology, for her
adviceandforproofreading.
AlanCurryacknowledgesthecontributionstohisworkofthepathol-
ogists, particularly Dr Helen Denley and Dr Lorna McWilliam, and
technical staff of the Manchester Royal Infirmary, as well as two inspi-
rational organisations – the Public Health Laboratory Service Electron
MicroscopynetworkandtheManchesterElectronMicroscopeSociety.
Brian Eyden wishes to thank all of the Pathology Department staff
at the Christie NHS Foundation Trust (Manchester), without whose
technicalandlightmicroscopicinputtheinterpretationoftumourultra-
structurewouldbecompromised,ifnot,insomeinstances,impossible.
Secondly,theeditorswishtorecognisethesupportandencouragement
of their families in this endeavour. John Stirling thanks his partner,
Jill, and expresses a special appreciation of his teachers and mentors,
particularly Alec Macfarlane who helped him achieve his dream of a
career in biology and Andrew Dorey who introduced him to electron
microscopy and the wonders of cell ultrastructure. Alan Curry thanks
vi ACKNOWLEDGEMENTSANDDEDICATION
his wife, Collette (particularly for her exceptional computer skills), and
Brian Eyden thanks his wife, Freda, for understanding the needs of a
writingscientist.
Finally, the editors dedicate this book to diagnostic electron micro-
scopists – wherever they may be – who continue to make uncertain
diagnoses more precise as a result of their labours, which, in turn, help
clinicianstotreattheirpatientsbetter,theultimatepurposeofourwork.
Contents
ListofContributors xvii
Preface – Introduction xxi
1 RenalDisease 1
JohnW.StirlingandAlanCurry
1.1 TheRoleofTransmissionElectronMicroscopy(TEM)
inRenalDiagnostics 1
1.2 UltrastructuralEvaluationandInterpretation 2
1.3 TheNormalGlomerulus 3
1.3.1 TheGlomerularBasementMembrane 4
1.4 UltrastructuralDiagnosticFeatures 5
1.4.1 Deposits:GeneralFeatures 5
1.4.2 GranularandAmorphousDeposits 6
1.4.3 OrganisedDeposits:FibrilsandTubules 7
1.4.4 NonspecificFibrils 11
1.4.5 GeneralandNonspecificInclusionsand
Deposits 11
1.4.6 Fibrin 12
1.4.7 TubuloreticularBodies(Tubuloreticular
Inclusions) 12
1.4.8 TheGlomerularBasementMembrane 13
1.4.9 TheMesangialMatrix 14
1.4.10 CellularComponentsoftheGlomerulus 14
1.4.11 ParietalEpithelium 16
1.5 TheUltrastructuralPathologyoftheMajor
GlomerularDiseases 16
1.5.1 Diseaseswithout,orwithOnlyMinor,
StructuralGBMChanges 16
1.5.2 DiseaseswithStructuralGBMChanges 19
viii CONTENTS
1.5.3 DiseaseswithGranularDeposits 25
1.5.4 DiseaseswithOrganisedDeposits 40
1.5.5 HereditaryMetabolicStorageDisorders 46
References 47
2 TransplantRenalBiopsies 55
JohnBrealey
2.1 Introduction 55
2.2 TheTransplantRenalBiopsy 55
2.3 IndicationsforElectronMicroscopyofTransplant
Kidney 56
2.3.1 TransplantGlomerulopathy 56
2.3.2 RecurrentPrimaryDisease 64
2.3.3 DeNovoGlomerularDisease 72
2.3.4 Donor-RelatedDisease 74
2.3.5 Infection 74
2.3.6 InconclusiveDiagnosisbyLMand/orIM 79
2.3.7 MiscellaneousTopics 81
References 84
3 ElectronMicroscopyinSkeletalMusclePathology 89
ElizabethCurtisandCarolineSewry
3.1 Introduction 89
3.1.1 TheBiopsyProcedure 90
3.1.2 Sampling 90
3.1.3 TissueProcessing 90
3.1.4 Artefacts 91
3.2 NormalMuscle 91
3.3 PathologicalChanges 96
3.3.1 Sarcolemma 96
3.3.2 Myofibrils 99
3.3.3 Glycogen 102
3.3.4 Cores 104
3.3.5 TargetFibres 105
3.3.6 Myonuclei 105
3.3.7 Mitochondria 106
3.3.8 ReticularSystem 108
3.3.9 Vacuoles 109
3.3.10 Capillaries 110
CONTENTS ix
3.3.11 OtherStructuralDefects 111
References 113
4 TheDiagnosticElectronMicroscopyofNerve 117
RosalindKing
4.1 Introduction 117
4.2 TissueProcessing 118
4.2.1 PreparationofNerveBiopsySpecimens 118
4.3 NormalNerveUltrastructure 120
4.3.1 Axons 120
4.3.2 SchwannCells 120
4.3.3 TheMyelinSheath 120
4.3.4 NodeofRanvier 122
4.3.5 Paranode 123
4.3.6 Juxtaparanode 123
4.3.7 Internode 123
4.3.8 Schmidt–LantermanIncisures 124
4.3.9 RemakFibres 124
4.3.10 Fibroblasts 124
4.3.11 RenautBodies 125
4.4 PathologicalUltrastructuralFeatures 125
4.4.1 AxonalDegeneration 125
4.4.2 AxonalRegeneration 126
4.4.3 RemakFibreAbnormalities 128
4.4.4 PolyglucosanBodies 128
4.4.5 NonspecificAxonalInclusions 128
4.4.6 DemyelinationandRemyelination 130
4.4.7 SpecificSchwannCellInclusions 135
4.4.8 NonspecificSchwannCellInclusions 136
4.4.9 Fibroblasts 142
4.4.10 PerineurialAbnormalities 142
4.4.11 CellularInfiltration 143
4.4.12 EndoneurialOedema 143
4.4.13 ConnectiveTissueAbnormalities 143
4.4.14 EndoneurialBloodVessels 145
4.4.15 MastCells 145
4.5 Artefact 145
4.6 Conclusions 147
References 148
x CONTENTS
5 TheDiagnosticElectronMicroscopyofTumours 153
BrianEyden
5.1 Introduction 153
5.2 PrinciplesandProceduresforDiagnosingTumoursby
ElectronMicroscopy 154
5.2.1 TheObjectiveofTumourDiagnosis 154
5.2.2 TheIntellectualRequirementsforTumour
DiagnosisbyElectronMicroscopy 155
5.2.3 TechnicalConsiderations 156
5.2.4 IdentifyingGoodPreservation 158
5.2.5 DistinguishingReactivefromNeoplasticCells 162
5.3 OrganellesandGroupsofCellStructuresDefining
CellularDifferentiation 162
5.3.1 RoughEndoplasmicReticulum 162
5.3.2 Melanosomes 165
5.3.3 Desmosomes 167
5.3.4 Tonofibrils 167
5.3.5 BasalLamina 169
5.3.6 GlandularEpithelialDifferentiationandCell
Processes 171
5.3.7 NeuroendocrineGranules 171
5.3.8 Smooth-MuscleMyofilaments 173
5.3.9 SarcomericMyofilaments(Thick-and-Thin
FilamentswithZ-Disks) 176
References 178
6 MicrobialUltrastructure 181
AlanCurry
6.1 Introduction 181
6.2 PracticalGuidance 182
6.3 Viruses 183
6.4 CurrentUseofEMinVirology 185
6.5 VirusesinThinSectionsofCellsorTissues 186
6.6 Bacteria 191
6.7 FungalOrganisms 194
6.8 Microsporidia 196
6.9 ParasiticProtozoa 206
6.9.1 Cryptosporidium 207
6.9.2 Isosporabelli 211
6.10 ExamplesofNon-entericProtozoa 212
