Physical Address

304 North Cardinal St.
Dorchester Center, MA 02124

Adjuvant Techniques–Immunohistochemistry, Cytogenetics, and Molecular Genetics

Immunohistochemistry Overview This section covers selected applications of immunohistochemistry (IHC) in the diagnosis of soft tissue and bone neoplasms, emphasizing the applications of IHC to the common differential diagnoses in soft tissue and bone pathology, including: (1) small, blue, round cell tumors; (2) monomorphic spindle cell tumors; (3) epithelioid tumors; and (4) pleomorphic spindle cell tumors. It is not possible in this brief section to provide…

Approach to Bone and Soft Tissue Tumors

Sarcomas are a heterogeneous group of rare mesenchymal malignant tumors. These tumors can arise in all sites of the body and are classified by their line of differentiation according to the type of mature tissue they resemble, either muscle, fat, fibrous tissue, bone, cartilage, blood vessels, or peripheral nerves. Benign mesenchymal tumors closely resemble normal tissue and have weak capability to invade local tissues, with little…

Mounting media and slide coatings

Introduction In order to provide the maximum degree of transparency to stained tissue sections, the refractive index of the mounting medium must approximate to that of dried protein, i.e. between 1.53 and 1.54. This is especially important for photographing slides. To visualize detail in unstained tissues, it may be desirable to employ a medium with lower or higher refractive index. The refractive index of a mounting…

Buffer solutions

Introduction The pH of a solution is defined as the logarithm to base 10 of 1 divided by the concentration of the free hydrogen ions in solution (i.e. pH = log 10 1/[A + ] = −log 10 [H + ]). A neutral solution is defined as pH = −log 10 [10 −7 ] = 7. The pH may greatly affect many chemical and immunohistochemical reactions,…

Preparation of solutions

Introduction Most solutions in histology are made using water as a solvent. The agent dissolved in water to make an aqueous solution is the solute. Solutions typically are made as volume to volume or weight to volume. Concentrated formaldehyde is a 40% solution of formaldehyde (CH 2 O) in water. This represents the maximum solubility of the molecule (solute), in water (the solvent) and the resulting…

Measurement units

Introduction SI Units (Système International d’Unités, ) are used throughout this text. Under ISO 15189 ( ) there are two standards relating to measurement and measurement units (ISO 15189 standards F3.3 and F3.4). To attain ISO 15189 accreditation, laboratories must ensure equipment used for critical measurements be calibrated to SI units and verified fit for purpose. In every test there is a degree of uncertainty but…

Applications of immunohistochemistry

Introduction The histological diagnosis is often clear from light microscopic examination of tinctorial-stained slides. However, in cases with varying differential diagnoses, or where molecular subtleties will add to case interpretation, immunohistochemistry may be employed to reach a conclusion. In certain circumstances, immunohistology also has a role in predicting prognosis and potential response to therapy. It is beyond the scope of this text to provide a comprehensive…

Tissue microarray

Introduction ∗ This is an abridged version of this topic in Chapter 6 of the 7 th edition of this text Tissue microarray (TMA) is a method used to evaluate numerous samples of tissue in a short time. First introduced by and further developed by it is used to examine several histological sections at the same time by arraying them in a single paraffin block. TMA…

Traditional methods

Introduction The aim of this appendix is to include discussion and methodology from the chapters Lipids, Proteins, Nucleic Acids and Enzyme Histochemistry which were included in previous editions of this book. These technologies have largely been superseded in the modern laboratory but are still used in teaching courses worldwide. For a full discussion and omitted methods please refer to the sixth edition of this text. Lipids…

Digital pathology

Key points Whole slide imaging (WSI) allows the creation of digital images of entire histology and cytology slides with sufficient detail to allow viewing at varying magnifications comparable to that achievable with a conventional light microscope. Acquiring digital WSIs requires dedicated hardware systems which create composite images of individually acquired tiles or lines. Such slide scanners typically include slide loading mechanisms, a motorized stage, a light…

Transmission electron microscopy

Acknowledgments We thank Richard Davey (SA Pathology) for his assistance in preparing the illustrations for this chapter and Dr Alan Curry (Manchester Royal Infirmary) for his helpful advice on microsporidia. Introduction Transmission electron microscopy (TEM) is a significant tool in demonstrating the ultrastructure of cells and tissues both in normal and disease states. In particular, TEM can be crucial in the diagnosis of various renal pathologies,…

Molecular pathology

Acknowledgment The authors acknowledge the use of some of the material and illustrations from the previous edition of this text. Introduction Tumor diagnosis relies centrally on careful morphological examination, supplemented by the judicious use of immunohistochemistry. The information gleaned by these means remains, without doubt, the single most important initial determinant of a patient’s prognosis and likely response to treatment. However, in recent years the importance…

Immunohistochemical and immunofluorescent techniques

Introduction The introduction of prognostic and predictive markers in immunohistochemistry (IHC) has made a tremendous beneficial impact on patient diagnosis and management. In the 1940s the demonstration of tissue constituents using an antibody labeled with a fluorescent dye was first described by . This original work used a method called direct immunofluorescence, where the specific primary antibody was conjugated directly with the fluorochrome and viewed under…

Neuropathology and muscle biopsy techniques

Acknowledgments Thanks to Sebastian Brandner for providing us with his protocol for methylene blue-azure II-basic fuchsin staining and to Patrick Elliott of the Medical Illustration Department, Royal Hallamshire Hospital, Sheffield, for his illustration depicted in Fig. 18.1 . We are grateful to Dr Gabriele De Luca for advice regarding applying the Palmgren method to cryostat sections, and to the neuropathology laboratories of the John Radcliffe Infirmary…

Bone

Acknowledgments The author would like to thank the following for their contributions to this chapter: Gayle Callis, for tape transfer photographs; and Chris Bull, for his expertise in CT and providing Fig. 17.14 . Introduction Bone is considered the most important supporting tissue in the body. It is composed of cells, organic extracellular matrix and inorganic salts. Bone tissue is mineralized in layers which provide great…

Traditional stains and modern techniques for demonstrating microorganisms in histology

Introduction Microorganisms (or microbes) are organisms which can only be individually seen by microscopy. Many do not cause disease in humans and act as normal colonizers of human hosts. Complex interactions between pathogens, which are capable of causing diseases, the host and the environment lead to clinical infections. Pathogens fall into five main groups: Viruses Bacteria Fungi Protozoa Helminths. With the advent of new and broader…

Amyloid

Acknowledgments Special thanks to friends, colleagues and patients at the National Amyloidosis Centre for their support and encouragement. Introduction History Amyloidosis is a disorder of protein folding, in which normally soluble proteins misfold and aggregate in a characteristic highly ordered fashion, and are deposited in the extracellular space as insoluble fibrils (or filaments). These interstitial fibrillar protein deposits (generically known as amyloid) may form anything from…

Pigments and minerals

Introduction In biology, pigments are defined as substances occurring in living matter which absorb visible light (electromagnetic energy within a narrow band which lies approximately between 400 and 800 nm). The various pigments may greatly differ in origin, chemical constitution and biological significance. They can be either organic or inorganic compounds which remain insoluble in most solvents. Minerals are naturally occurring homogeneous, inorganic substances having a…

Carbohydrates

Introduction This is a large group of compounds with the general formula C n (H 2 O) n . The role of carbohydrates in cellular metabolism has been known for many years but carbohydrates have more recently been implicated in a wide range of cellular functions including protein folding, cell adhesion, enzyme activity and immune recognition ( ). Histochemical techniques for the detection and characterization of…