Diagnostic Electron Microscopy

Diagnostic Electron Microscopy: A Practical Guide Interpretation and Technique summarises the current interpretational applications of TEM in diagnostic pathology. This concise and accessible volume provides a working guide to the main, or most useful, applications of the technique including practical topics of concern to laboratory scientists, brief guides to traditional tissue and microbiological preparation techniques, microwave processing, digital imaging and measurement uncertainty. The text features both a screening and interpretational guide for TEM diagnostic applications and current TEM diagnostic tissue preparation methods pertinent to all clinical electron microscope units worldwide. Containing high-quality representative images, this up-to-date text includes detailed information on the most important diagnostic applications of transmission electron microscopy as well as instructions for specific tissues and current basic preparative techniques. The book is relevant to trainee pathologists and practising pathologists who are expected to understand and evaluate/screen tissues by TEM. In addition, technical and scientific staff involved in tissue preparation and diagnostic tissue evaluation/screening by TEM will find this text useful. CONTENTS 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