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Diagnostic Electron Microscopy: A Practical Guide to 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.

Diagnostic Electron Microscopy: A Practical Guide to Tissue Preparation and Interpretation

List of Contributors xxiiiPreface xxvii1 Renal Disease 1John W. Stirling and Alan Curry1.1 The Role of Transmission Electron Microscopy (TEM) in Renal Diagnostics 11.2 Ultrastructural Evaluation and Interpretation 21.3 The Normal Glomerulus 31.4 Ultrastructural Diagnostic Features 51.5 Granular and Amorphous Deposits 61.6 Organised Immune-Related Fibrils and Tubules 71.7 Nonspecific Fibrils 111.8 General and Nonspecific Inclusions and Deposits 111.9 Fibrin 121.10 Tubuloreticular Bodies (Tubuloreticular Inclusions) 121.11 The Glomerular Basement Membrane 131.12 The Mesangial Matrix 141.13 Cellular Components of the Glomerulus 141.14 Mesangial Cells 161.15 The Capillary Lumen 161.16 The Ultrastructural Pathology of the Major Glomerular Diseases 161.17 Focal Segmental Glomerulosclerosis 171.18 Pauci-Immune Glomerulonephritis (ANCA-Associated Glomerulonephritis) 181.19 Anti-GBM Glomerulonephritis (Goodpasture Syndrome) 191.20 Diseases with Structural GBM Changes 191.21 Thin Basement Membrane Disease 211.22 Diabetic Nephropathy 221.23 Thrombotic Microangiopathy: Haemolytic Uraemic Syndrome and Thrombotic ThrombocytopaenicPurpura 241.24 Diseases with Granular Deposits 251.25 Acute Post-infectious Glomerulonephritis 291.26 IgA Nephropathy (Berger's Disease) and Henoch-Schonlein Purpura Nephritis 301.27 Mesangiocapillary Glomerulonephritis (Membranoproliferative Glomerulonephritis) 321.28 Systemic Lupus Erythematosus 361.29 Monoclonal Immunoglobulin Deposition Disease 371.30 Diseases with Organised Deposits 401.31 Fibrillary Glomerulonephritis 411.32 Immunotactoid Glomerulonephritis (Immunotactoid GN) 421.33 Cryoglobulinaemic Glomerulonephritis 451.34 Hereditary Metabolic Storage Disorders 461.35 Fabry's Disease 461.36 Lecithin Cholesterol Acyltransferase Deficiency 47References 472 Diagnostic Transmission Electron Microscopy of Transplant Renal Biopsies 55John Brealey2.1 Introduction 552.2 The Transplant Renal Biopsy 562.3 Indications for Electron Microscopy of Transplant Kidney 56References 843 The Use of Electron Microscopy in Skeletal Muscle Pathology 89Elizabeth Curtis and Caroline Sewry3.1 Introduction 893.2 Normal Muscle 913.3 Pathological Changes 96References 1134 The Diagnostic Electron Microscopy of Nerve 117Rosalind King4.1 Introduction 1174.2 Tissue Processing 1184.3 Normal Nerve Ultrastructure 1204.4 Pathological Ultrastructural Features 1254.5 Artefact 1454.6 Conclusions 147References 1485 The Diagnostic Electron Microscopy of Tumours 153Brian Eyden5.1 Introduction 1535.2 Principles and Procedures for Diagnosing Tumours by Electron Microscopy 1545.3 Organelles and Groups of Cell Structures Defining Cellular Differentiation 162References 1786 Microbial Ultrastructure 181Alan Curry6.1 Introduction 1816.2 Practical Guidance 1826.3 Viruses 1836.4 Current Use of EM in Virology 1856.5 Viruses in Thin Sections of Cells or Tissues 1866.6 Bacteria 1916.7 Fungal Organisms 1946.8 Microsporidia 1966.9 Parasitic Protozoa 2066.10 Examples of Non-enteric Protozoa 2126.11 Parasitic Amoebae 2136.12 Conclusions 214Acknowledgements 214References and Additional Reading 2147 The Contemporary Use of Electron Microscopy in the Diagnosis of Ciliary Disorders and Sperm Centriolar Abnormalities 221P. Yiallouros, M. Nearchou, A. Hadjisavvas and K. Kyriacou7.1 Introduction 2217.2 Ultrastructure of Motile Cilia 2247.3 Genetics of PCD 2267.4 Current Diagnostic Modalities 2287.5 Clinical Features 2297.6 Procurement and Assessment of Ciliated Specimens 2307.7 Centriolar Sperm Abnormalities 2317.8 Discussion 232Acknowledgements 234References 2348 Electron Microscopy as a Useful Tool in the Diagnosis of Lysosomal Storage Diseases 237Joseph Alroy, Rolf Pfannl, and Angelo A. Ucci8.1 Introduction 2378.2 Morphological Findings 2478.3 Conclusion 261References 2629 Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) 269John W. Stirling9.1 Introduction 2699.2 Diagnostic Strategies - Comparative Specificity and Sensitivity 2719.3 Diagnosis by TEM 271References 27410 Diagnosis of Platelet Disorders by Electron Microscopy 277Hilary Christensen and Walter H.A. Kahr10.1 Introduction 27710.2 TEM Preparation of Platelets 27810.3 Whole-Mount EM Preparation of Platelets 28010.4 EM Preparation of Bone Marrow 28110.5 Pre-embed Immunogold Labelling of Von Willibrand Factor in Platelets 28210.6 Ultrastructural Features of Platelets 28210.7 Normal Platelets 28310.8 Ultrastructural Characteristics of Abnormal Platelets: Grey Platelet Syndrome 28510.9 Arthrogryposis, Renal Dysfunction and Cholestasis Syndrome 28510.10 Jacobsen Syndrome 28610.11 Hermansky-Pudlak Syndrome, Chediak-Higashi Syndrome and Other Dense-Granule Deficiencies 28710.12 Type 2B von Willebrand Disease and Platelet-Type von Willebrand Disease 288References 29011 Diagnosis of Congenital Dyserythropoietic Anaemia Types I and II by Transmission Electron Microscopy 293Yong-xin Ru11.1 Introduction 29311.2 Preparation of Bone Marrow and General Observation Protocol 29411.3 CDA Type I 29411.4 CDA Type II 29911.5 Summary 306Acknowledgements 307References 30712 Ehlers-Danlos Syndrome 309Trinh Hermanns-Le, Marie-Annick Reginster, Claudine Pierard-Franchimont and Gerald E. Pierard12.1 Introduction 30912.2 Collagen Fibrils 31012.3 Elastic Fibers 31012.4 Nonfibrous Stroma and Granulo-Filamentous Deposits 31112.5 Connective Tissue Disorders 311References 31913 Electron Microscopy in Occupational and Environmental Lung Disease 323Victor L. Roggli13.1 Introduction 32313.2 Asbestos 32413.3 Hypersensitivity Pneumonitis and Sarcoidosis 33013.4 Silicosis 33113.5 Silicate Pneumoconiosis 33313.6 Metal-Induced Diseases 33513.7 Rare-Earth Pneumoconiosis 33813.8 Miscellaneous Disorders 338References 33914 General Tissue Preparation Methods 341John W. Stirling14.1 Introduction 34114.2 Tissue Collection and Dissection 34214.3 Tissue Processing 34514.4 Tissue Sectioning 352References 37915 Ultrastructural Pathology Today - Paradigm Change and the Impact of Microwave Technology and Telemicroscopy 383Josef A. Schroeder15.1 Diagnostic Electron Microscopy and Paradigm Shift in Pathology 38315.2 Standardised and Automated Conventional Tissue Processing 38515.3 Microwave-Assisted Sample Preparation#16 39015.4 Cyberspace for Telepathology via the Internet 39715.5 Conclusions and Future Prospects 400Acknowledgements 404References 40416 Electron Microscopy Methods in Virology 409Alan Curry16.1 Biological Safety Precautions 40916.2 Collection of Specimens 41016.3 Preparation of Faeces, Vomitus or Urine Samples 41016.4 Viruses in Skin Lesions 41016.5 Reagents and Methods 41116.6 Coated Grids 41216.7 Important Elements in the Negative Staining Procedure 41216.8 TEM Examination 41316.9 Immunoelectron Microscopy 41316.10 Thin Sectioning of Virus-Infected Cells or Tissues 41416.11 Virology Quality Assurance (QA) Procedures 415Acknowledgements 415References 41617 Digital Imaging for Diagnostic Transmission Electron Microscopy 419Gary Paul Edwards17.1 Introduction 41917.2 Camera History 41917.3 The Pixel Dilemma 42017.4 Camera Positioning 42117.5 Resolution 42217.6 Fibre Coupled or Lens Coupled? 42317.7 Sensitivity, Noise and Dynamic Range 42417.8 CCD Chip Type (Full Frame or Interline) 42617.9 Binning and Frame Rate 42617.10 Software 42717.11 Choosing the Right Camera 428References 42918 Uncertainty of Measurement 431Pierre Filion18.1 Introduction 43118.2 Purpose 43218.3 Factors That Influence Quantitative Measurements 43318.4 How to Calculate the UM 44018.5 Worked Examples 44418.6 Conclusion 446References 447Index 449