List of Contributors xxiii
Preface xxvii
1 Renal Disease 1
John W. Stirling and Alan Curry
1.1 The Role of Transmission Electron Microscopy (TEM) in Renal Diagnostics 1
1.2 Ultrastructural Evaluation and Interpretation 2
1.3 The Normal Glomerulus 3
1.4 Ultrastructural Diagnostic Features 5
1.5 Granular and Amorphous Deposits 6
1.6 Organised Immune-Related Fibrils and Tubules 7
1.7 Nonspecific Fibrils 11
1.8 General and Nonspecific Inclusions and Deposits 11
1.9 Fibrin 12
1.10 Tubuloreticular Bodies (Tubuloreticular Inclusions) 12
1.11 The Glomerular Basement Membrane 13
1.12 The Mesangial Matrix 14
1.13 Cellular Components of the Glomerulus 14
1.14 Mesangial Cells 16
1.15 The Capillary Lumen 16
1.16 The Ultrastructural Pathology of the Major Glomerular Diseases 16
1.17 Focal Segmental Glomerulosclerosis 17
1.18 Pauci-Immune Glomerulonephritis (ANCA-Associated Glomerulonephritis) 18
1.19 Anti-GBM Glomerulonephritis (Goodpasture Syndrome) 19
1.20 Diseases with Structural GBM Changes 19
1.21 Thin Basement Membrane Disease 21
1.22 Diabetic Nephropathy 22
1.23 Thrombotic Microangiopathy: Haemolytic Uraemic Syndrome and Thrombotic Thrombocytopaenic
Purpura 24
1.24 Diseases with Granular Deposits 25
1.25 Acute Post-infectious Glomerulonephritis 29
1.26 IgA Nephropathy (Berger’s Disease) and Henoch-Schonlein Purpura Nephritis 30
1.27 Mesangiocapillary Glomerulonephritis (Membranoproliferative Glomerulonephritis) 32
1.28 Systemic Lupus Erythematosus 36
1.29 Monoclonal Immunoglobulin Deposition Disease 37
1.30 Diseases with Organised Deposits 40
1.31 Fibrillary Glomerulonephritis 41
1.32 Immunotactoid Glomerulonephritis (Immunotactoid GN) 42
1.33 Cryoglobulinaemic Glomerulonephritis 45
1.34 Hereditary Metabolic Storage Disorders 46
1.35 Fabry’s Disease 46
1.36 Lecithin Cholesterol Acyltransferase Deficiency 47
References 47
2 Diagnostic Transmission Electron Microscopy of Transplant Renal Biopsies 55
John Brealey
2.1 Introduction 55
2.2 The Transplant Renal Biopsy 56
2.3 Indications for Electron Microscopy of Transplant Kidney 56
References 84
3 The Use of Electron Microscopy in Skeletal Muscle Pathology 89
Elizabeth Curtis and Caroline Sewry
3.1 Introduction 89
3.2 Normal Muscle 91
3.3 Pathological Changes 96
References 113
4 The Diagnostic Electron Microscopy of Nerve 117
Rosalind King
4.1 Introduction 117
4.2 Tissue Processing 118
4.3 Normal Nerve Ultrastructure 120
4.4 Pathological Ultrastructural Features 125
4.5 Artefact 145
4.6 Conclusions 147
References 148
5 The Diagnostic Electron Microscopy of Tumours 153
Brian Eyden
5.1 Introduction 153
5.2 Principles and Procedures for Diagnosing Tumours by Electron Microscopy 154
5.3 Organelles and Groups of Cell Structures Defining Cellular Differentiation 162
References 178
6 Microbial Ultrastructure 181
Alan Curry
6.1 Introduction 181
6.2 Practical Guidance 182
6.3 Viruses 183
6.4 Current Use of EM in Virology 185
6.5 Viruses in Thin Sections of Cells or Tissues 186
6.6 Bacteria 191
6.7 Fungal Organisms 194
6.8 Microsporidia 196
6.9 Parasitic Protozoa 206
6.10 Examples of Non-enteric Protozoa 212
6.11 Parasitic Amoebae 213
6.12 Conclusions 214
Acknowledgements 214
References and Additional Reading 214
7 The Contemporary Use of Electron Microscopy in the Diagnosis of Ciliary Disorders and Sperm Centriolar Abnormalities 221
P. Yiallouros, M. Nearchou, A. Hadjisavvas and K. Kyriacou
7.1 Introduction 221
7.2 Ultrastructure of Motile Cilia 224
7.3 Genetics of PCD 226
7.4 Current Diagnostic Modalities 228
7.5 Clinical Features 229
7.6 Procurement and Assessment of Ciliated Specimens 230
7.7 Centriolar Sperm Abnormalities 231
7.8 Discussion 232
Acknowledgements 234
References 234
8 Electron Microscopy as a Useful Tool in the Diagnosis of Lysosomal Storage Diseases 237
Joseph Alroy, Rolf Pfannl, and Angelo A. Ucci
8.1 Introduction 237
8.2 Morphological Findings 247
8.3 Conclusion 261
References 262
9 Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) 269
John W. Stirling
9.1 Introduction 269
9.2 Diagnostic Strategies – Comparative Specificity and Sensitivity 271
9.3 Diagnosis by TEM 271
References 274
10 Diagnosis of Platelet Disorders by Electron Microscopy 277
Hilary Christensen and Walter H.A. Kahr
10.1 Introduction 277
10.2 TEM Preparation of Platelets 278
10.3 Whole-Mount EM Preparation of Platelets 280
10.4 EM Preparation of Bone Marrow 281
10.5 Pre-embed Immunogold Labelling of Von Willibrand Factor in Platelets 282
10.6 Ultrastructural Features of Platelets 282
10.7 Normal Platelets 283
10.8 Ultrastructural Characteristics of Abnormal Platelets: Grey Platelet Syndrome 285
10.9 Arthrogryposis, Renal Dysfunction and Cholestasis Syndrome 285
10.10 Jacobsen Syndrome 286
10.11 Hermansky–Pudlak Syndrome, Chediak–Higashi Syndrome and Other Dense-Granule Deficiencies 287
10.12 Type 2B von Willebrand Disease and Platelet-Type von Willebrand Disease 288
References 290
11 Diagnosis of Congenital Dyserythropoietic Anaemia Types I and II by Transmission Electron Microscopy 293
Yong-xin Ru
11.1 Introduction 293
11.2 Preparation of Bone Marrow and General Observation Protocol 294
11.3 CDA Type I 294
11.4 CDA Type II 299
11.5 Summary 306
Acknowledgements 307
References 307
12 Ehlers–Danlos Syndrome 309
Trinh Hermanns-Le, Marie-Annick Reginster, Claudine Pierard-Franchimont and Gerald E. Pierard
12.1 Introduction 309
12.2 Collagen Fibrils 310
12.3 Elastic Fibers 310
12.4 Nonfibrous Stroma and Granulo-Filamentous Deposits 311
12.5 Connective Tissue Disorders 311
References 319
13 Electron Microscopy in Occupational and Environmental Lung Disease 323
Victor L. Roggli
13.1 Introduction 323
13.2 Asbestos 324
13.3 Hypersensitivity Pneumonitis and Sarcoidosis 330
13.4 Silicosis 331
13.5 Silicate Pneumoconiosis 333
13.6 Metal-Induced Diseases 335
13.7 Rare-Earth Pneumoconiosis 338
13.8 Miscellaneous Disorders 338
References 339
14 General Tissue Preparation Methods 341
John W. Stirling
14.1 Introduction 341
14.2 Tissue Collection and Dissection 342
14.3 Tissue Processing 345
14.4 Tissue Sectioning 352
References 379
15 Ultrastructural Pathology Today – Paradigm Change and the Impact of Microwave Technology and Telemicroscopy 383
Josef A. Schroeder
15.1 Diagnostic Electron Microscopy and Paradigm Shift in Pathology 383
15.2 Standardised and Automated Conventional Tissue Processing 385
15.3 Microwave-Assisted Sample Preparation#16 390
15.4 Cyberspace for Telepathology via the Internet 397
15.5 Conclusions and Future Prospects 400
Acknowledgements 404
References 404
16 Electron Microscopy Methods in Virology 409
Alan Curry
16.1 Biological Safety Precautions 409
16.2 Collection of Specimens 410
16.3 Preparation of Faeces, Vomitus or Urine Samples 410
16.4 Viruses in Skin Lesions 410
16.5 Reagents and Methods 411
16.6 Coated Grids 412
16.7 Important Elements in the Negative Staining Procedure 412
16.8 TEM Examination 413
16.9 Immunoelectron Microscopy 413
16.10 Thin Sectioning of Virus-Infected Cells or Tissues 414
16.11 Virology Quality Assurance (QA) Procedures 415
Acknowledgements 415
References 416
17 Digital Imaging for Diagnostic Transmission Electron Microscopy 419
Gary Paul Edwards
17.1 Introduction 419
17.2 Camera History 419
17.3 The Pixel Dilemma 420
17.4 Camera Positioning 421
17.5 Resolution 422
17.6 Fibre Coupled or Lens Coupled? 423
17.7 Sensitivity, Noise and Dynamic Range 424
17.8 CCD Chip Type (Full Frame or Interline) 426
17.9 Binning and Frame Rate 426
17.10 Software 427
17.11 Choosing the Right Camera 428
References 429
18 Uncertainty of Measurement 431
Pierre Filion
18.1 Introduction 431
18.2 Purpose 432
18.3 Factors That Influence Quantitative Measurements 433
18.4 How to Calculate the UM 440
18.5 Worked Examples 444
18.6 Conclusion 446
References 447
Index 449
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