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

John Stirling (Editor), Alan Curry (Editor), Brian Eyden (Editor)
ISBN: 978-1-119-97399-7
492 pages
January 2013
Diagnostic Electron Microscopy: A Practical Guide to Tissue Preparation and Interpretation (1119973996) cover image

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.
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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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“Thus, this book is a “must-have” for all pathology departments, even if they are not equipped with an EM facility, and it is also a solid proof of the current role of electron microscopy in health care.”  (Microscopy & Microanalysis, 1 August 2013)

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