Histotechnology: A Self-Instructional Text

Description

An indispensable teaching tool and reference--and a "must" for histotechnologists preparing for the ASCP HTL certifying examination--Histotechnology has been completely updated in the new 3rd edition. New coverage includes chapters on immunohistochemistry and molecular techniques and cytopreparation. You'll also appreciate new features incorporated throughout the book--new images, how-to illustrations for manual techniques, troubleshooting aids, and additional special staining procedures. Cover Page Title Page Table of Contents Preface Chapter 1: Fixation Definition Functions of Fixatives Actions of Fixatives Factors Affecting Fixation TEMPERATURE SIZE VOLUME RATIO TIME CHOICE OF FIXATIVE PENETRATION TISSUE STORAGE pH OSMOLALITY Reactions of the Cell with Fixatives THE NUCLEUS PROTEINS LIPIDS CARBOHYDRATES Simple Aqueous Fixatives or Fixative Ingredients ACETIC ACID FORMALDEHYDE GLUTARALDEHYDE GLYOXAL MERCURIC CHLORIDE OSMIUM TETROXIDE PICRIC ACID POTASSIUM DICHROMATE ZINC SALTS Other Fixative Ingredients Compound or Combined Fixatives B-5 FIXATIVE ZENKER AND HELLY (ZENKER-FORMOL) SOLUTIONS ZINC FORMALIN SOLUTIONS Nonaqueous Fixatives ACETONE ALCOHOL Transport Solutions Removal of Fixation Pigments Troubleshooting Fixation Problems AUTOLYSIS INCOMPLETE FIXATION References LEARNING ACTIVITIES Chapter 2: Processing Dehydration ALCOHOLS UNIVERSAL SOLVENTS Clearing XYLENE TOLUENE BENZENE CHLOROFORM ACETONE ESSENTIAL OILS LIMONENE REAGENTS (XYLENE SUBSTITUTE) ALIPHATIC HYDROCARBONS (XYLENE SUBSTITUTE) UNIVERSAL SOLVENTS OTHER CLEARING AGENTS Infiltration PARAFFIN WATER-SOLUBLE WAXES CELLOIDIN PLASTICS AGAR AND GELATIN 30% SUCROSE Troubleshooting Processing PRECIPITATE IN THE PROCESSOR CHAMBER AND INTHE TUBING OVERDEHYDRATION POOR PROCESSING SPONGE ARTIFACT TISSUE ACCIDENTALLY DESICCATED Embedding and Specimen Orientation Troubleshooting Embedding SOFT MUSHY TISSUE INCORRECT ORIENTATION TISSUE CARRYOVER TISSUE NOT EMBEDDED AT THE SAME LEVEL PIECES OF TISSUE MISSING FROM THE BLOCK Special Techniques in Processing DECALCIFICATION TROUBLESHOOTING DECALCIFICATION FROZEN SECTIONS TROUBLESHOOTING PROCESSING TISSUE FORFROZEN SECTIONS References LEARNING ACTIVITIES Chapter 3: Instrumentation Microscopes LIGHT MICROSCOPE POLARIZING MICROSCOPE PHASE-CONTRAST MICROSCOPE DARKFIELD MICROSCOPE FLUORESCENCE MICROSCOPE ELECTRON MICROSCOPE Microtomes ROTARY MICROTOME SLIDING MICROTOME CLINICAL FREEZING MICROTOME MICROTOME BLADES TROUBLESHOOTING MICROTOMY Cryostat Tissue Processors CONVENTIONAL PROCESSOR MICROWAVE PROCESSOR Stainers and Coverslippers MICROWAVE STAINING OVEN AUTOMATIC COVERSLIPPER Miscellaneous Equipment FLOTATION BATHS CHROMIUM POTASSIUM SULFATE-COATED SLIDES POLY-L-LYSINE-COATED SLIDES AMINOALKYLSILANE-TREATED SLIDES [RENTROP 1986] DRYERS AND OVENS CIRCULATING WATER BATH FREEZERS AND REFRIGERATORS pH METERS BALANCES AND SCALES EMBEDDING CENTER MICROMETER PIPETTES SOLVENT RECYCLER Instrument Quality Control NEW INSTRUMENT VALIDATION QUALITY CONTROL PROGRAM References Equipment Temperature Quality Control Chart Equipment Maintenance and History Chapter 4: Safety Biological or Infectious Hazards TUBERCULOSIS EXPOSURE CRYOGENIC SPRAYS HIV, HEPATITIS c VIRUS (Hcv), AND HBV CREUTZFELDT-JAKOB DISEASE (CJD) HANDLING TISSUE WASTE Mechanical Hazards ERGONOMICS Chemical Hazards PARTICULARLY HAZARDOUS SUBSTANCES(REPRODUCTIVE TOXINS, SELECT CARCINOGENS,AND SUBSTANCES WITH A HIGH DEGREE OF ACUTETOXICITY) CARCINOGENS CORROSIVE SUBSTANCES FIRE AND EXPLOSION HAZARDS HAZARDOUS CHEMICAL SPILLS AND STORAGE CHEMICAL STORAGE HAZARDOUS CHEMICAL DISPOSAL Hazard Identification General Safety Practices EMPLOYEES: SUPERVISORS: References LEARNING ACTIVITIES Chapter 5: Laboratory Mathematics and Solution Preparation Percentage Solutions Use of the Gravimetric Factor in Solution Preparation Hydrates Normal and Molar Solutions The Metric System TEMPERATURE CONVERSION Buffers General Guidelines for Solution Preparation, Use, and Storage Stability of Solutions References ANSWERS TO PROBLEMS IN CHAPTER ANSWERS TO PROBLEMS IN LEARNING ACTIVITIES LEARNING ACTIVITIES Chapter 6: Nuclear and Cytoplasmic Staining Ultrastructure of the Cell THE NUCLEUS THE CYTOPLASM Staining Mechanisms NUCLEAR STAINING CYTOPLASMIC STAINING The Dyes FACTORS AFFECTING DYE BINDING DIFFERENTIATION THE NUCLEAR DYES PLASMA STAINS H&E Staining MANUAL PROGRESSIVE STAINING METHOD MANUAL REGRESSIVE STAINING METHOD AUTOMATED STAINING FROZEN SECTION STAINING Troubleshooting the H&E Stain INCOMPLETE DEPARAFFINIZATION NUCLEAR STAINING IS NOT CRISP PALE NUCLEAR STAINING DARK NUCLEAR STAINING RED OR RED-BROWN NUCLEI PALE CYTOPLASMIC STAINING DARK CYTOPLASMIC STAINING EOSIN NOT PROPERLY DIFFERENTIATED BLUE-BLACK PRECIPITATE ON TOP OF SECTIONS HAZY OR MILKY WATER AND SLIDES UNEVEN H&E STAINING DARK BASOPHILIC STAINING OF NUCLEI ANDCYTOPLASM, ESPECIALLY AROUND TISSUE EDGES POOR CONTRAST BETWEEN NUCLEUS ANDCYTOPLASM Nucleic Acid Stains FEULGEN REACTION METHYL GREEN-PYRONIN Y STOCK ACETATE BUFFER SOLUTIONS Polychromatic Stains MAY-GRUNWALD GIEMSA STAIN Mounting Stained Sections RESINOUS MEDIA AQUEOUS MOUNTING MEDIA COVERSLIPS Troubleshooting Mounted Stained Sections WATER BUBBLES NOTED IN MOUNTED SECTIONS ALL AREAS OF SECTION CANNOT BE BROUGHTINTO FOCUS CORN-FLAKING ARTIFACT SEEN ONMOUNTED SECTIONS MOUNTED STAINED SECTIONS ARE NOT AS CRISPAS USUAL WHEN VIEWED MICROSCOPICALLY RETRACTED MOUNTING MEDIUM References LEARNING ACTIVITIES Chapter 7: Carbohydrates and Amyloid Carbohydrates GROUP 1: NEUTRAL POLYSACCHARIDES (NONIONICHOMOGLYCANS) GROUP II: ACID MUCOPOLYSACCHARIDES (ANIONICHETEROGLYCANS) GROUP III: GLYCOPROTEINS (MUCINS, MUCOID,MUCOPROTEIN, MUCOSUBSTANCES) GROUP IV: GLYCOLIPIDS Special Staining Techniques PAS REACTION TEST FOR QUALITY OF SCHIFF REAGENT PAS REACTION WITH DIASTASE DIGESTION TEST FOR QUALITY OF SCHIFF REAGENT BEST CARMINE MAYER MUCICARMINE ALCIAN BLUE, pH 2.5 ALCIAN BLUE, pH 1.0 ALCIAN BLUE WITH HYALURONIDASE ALCIAN BLUE-PAS-HEMATOXYLIN MÜLLER-MOWRY COLLOIDAL IRON Amyloid ALKALINE CONGO RED METHOD CRYSTAL VIOLET THIOFLAVINE T FLUORESCENT METHOD References LEARNING ACTIVITIES Chapter 8: Connective and Muscle Tissue Connective Tissue Basement Membrane Muscle Staining Techniques for Connective Tissue Fibers MASSON TRICHROME STAIN [ GOMORI 1-STEP TRICHROME STAIN VANGIESON PICRIC ACID-ACID FUCHSIN STAIN VERHOEFF ELASTIC STAIN ALDEHYDE FUCHSIN ELASTIC STAIN NOTES ON OTHER ELASTIC STAINS RUSSELL MODIFICATION OF THE MOVATPENTACHROME STAIN SILVER TECHNIQUES FOR RETICULAR FIBERS GOMORI STAIN FOR RETICULAR FIBERS GORDON AND SWEETS STAIN FOR RETICULARFIBERS Staining Techniques for Muscle MALLORY PTAH TECHNIQUE FOR CROSS-STRIATIONSAND FIBRIN PTAH WITHOUT MERCURIC SOLUTIONS Staining Technique for Basement Membranes PERIODIC ACID-METHENAMINE SILVER MICROWAVEPROCEDURE FOR BASEMENT MEMBRANES Staining Techniques for Lipid OIL RED O METHOD FOR NEUTRAL FATS SUDAN BLACK B IN PROPYLENE GLYCOL OSMIUM TETROXIDE PARAFFIN PROCEDURE FORFAT Staining Techniques for Connective Tissue Cells TOLUIDINE BLUE FOR MAST CELLS METHYL GREEN-PYRONIN Y References LEARNING ACTIVITIES Chapter 9: Nerve The Nervous System Neurons NISSL SUBSTANCE NERVE CELL PROCESSES Neuroglia OLIGODENDROGLIA ASTROCYTES MICROGLIA EPENDYMAL CELLS Myelin Special Staining Techniques NISSL SUBSTANCE: CRESYL ECHT VIOLET METHOD I NISSL SUBSTANCE: CRESYL ECHT VIOLET METHOD II NERVE FIBERS, NERVE ENDINGS, NEUROFIBRILS:BODIAN METHOD NERVE FIBERS AND NEUROFIBRILS: HOLMES SILVERNITRATE METHOD NERVE FIBERS, NEUROFIBRILLARY TANGLES, ANDSENILE PLAQUES: BIELSCHOWSKY-PAS STAIN NERVE FIBERS, NEUROFIBRILLARY TANGLES, ANDSENILE PLAQUES: MICROWAVE MODIFICATION OFBIELSCHOWSKY METHOD NERVE FIBERS, NEUROFIBRILLARY TANGLES,AND SENILE PLAQUES: T HE SEVIER-MUNGERMODIFICATION OF BIELSCHOWSKY METHOD NEUROFIBRILLARY TANGLES AND SENILE PLAQUES:THIOFLAVIN S (MODIFIED) GLIAL FIBERS: MALLORY PHOSPHOTUNGSTIC ACIDHEMATOXYLIN (PTAH) STAIN GLIAL FIBERS: HOLZER METHOD ASTROCYTES: CAJAL STAIN MYELIN SHEATH: WEIL METHOD MYELIN SHEATH: LUXOL FAST BLUE METHOD MYELIN SHEATH AND NISSL SUBSTANCE COMBINED:LUXOL FAST BLUE-CRESYL ECHT VIOLET STAIN MYELIN SHEATHS AND NERVE FIBERS COMBINED:LUXOL FAST BLUE-HOLMES SILVER NITRATE METHOD LUXOL FAST BLUE-PAS-HEMATOXYLIN References LEARNING ACTIVITIES Chapter 10: Microorganisms Bacteria Fungi Virsues Protozoans Special Staining Techniques KINYOUN ACID-FAST STAIN ZIEHL-NEELSEN METHOD FOR ACID-FAST BACTERIA(AFIP MODIFICATION) MICROWAVE ZIEHL-NEELSEN METHOD FOR ACIDFASTBACTERIA FITE ACID-FAST STAIN FOR LEPROSY ORGANISMS MICROWAVE AURAMINE-RHODAMINE FLUORESCENCETECHNIQUE BROWN-HOPPS MODIFICATION OF THE GRAM STAIN GIEMSA METHODS MODIFIED DIFF-QUIK GIEMSA STAIN FORHELICOBACTER PYLORI ALCIAN YELLOW-TOLUIDINE BLUE METHOD FOR HPYLORI HOTCHKISS-MCMANUS PAS REACTION FOR FUNGI CHROMIC ACID-SCHIFF STAIN FOR FUNGI (CAS) GRIDLEY FUNGUS STAIN GROCOTT METHENAMINE-SILVER NITRATE FUNGUSSTAIN MICROWAVE METHENAMINE-SILVER NITRATEPROCEDURE FOR FUNGI MAYER MUCICARMINE AND ALCIAN BLUETECHNIQUES FOR CRYPTOCOCCUS NEOFORMANS WARTHIN-STARRY TECHNIQUE FOR SPIROCHETES MICROWAVE MODIFICATION OF THE WARTHINSTARRYMETHOD FOR BACTERIA DIETERLE METHOD FOR SPIROCHETES ANDLEGIONELLA ORGANISMS MICROWAVE STEINER AND STEINER PROCEDUREFOR SPIROCHETES, HELICOBACTER, AND LEGIONELLAORGANISMS References LEARNING ACTIVITIES Chapter 11: Pigments, Minerals, and Cytoplasmic Granules Pigments ARTIFACT PIGMENTS EXOGENOUS PIGMENTS ENDOGENOUS HEMATOGENOUS PIGMENTS ENDOGENOUS NONHEMATOGENOUS PIGMENT Endogenous Deposits Minerals Cytoplasmic Granules Special Staining Techniques PRUSSIAN BLUE STAIN FOR FERRIC IRON TURNBULL BLUE STAIN FOR FERROUS IRON SCHMORL TECHNIQUE FOR REDUCING SUBSTANCES FONTANA-MASSON STAIN FOR MELANIN ANDARGENTAFFIN GRANULES MICROWAVE FONTANA-MASSON STAIN GRIMELIUS ARGYROPHIL STAIN CHURUKIAN-SCHENK METHOD FOR ARGYROPHILGRANULES MICROWAVE CHURUKIAN-SCHENK METHOD FORARGYROPHIL GRANULES GOMORI METHENAMINE-SILVER METHOD FORURATES BILE STAIN VON KOSSA CALCIUM STAIN ALIZARIN RED S CALCIUM STAIN RHODANINE METHOD FOR COPPER MICROWAVE RHODANINE COPPER METHOD References LEARNING ACTIVITIES Chapter 12: Immunohistochemistry Introduction General Immunology ANTIBODY ANTIGEN POLYCLONAL ANTISERA MONOCLONAL ANTIBODIES RABBIT MONOCLONAL ANTIBODIES Tissue Handling FROZEN TISSUE FIXATION AND PROCESSING FIXATIVES FOR PARAFFIN-PROCESSED TISSUE PROCESSING MICROTOMY EPITOPE ENHANCEMENT OR RETRIEVAL Methods of Visualization ENZYME IMMUNOHISTOCHEMISTRY Immunohistochemical Staining Methods DIRECT METHOD INDIRECT METHOD UNLABELED, OR SOLUBLE ENZYME IMMUNECOMPLEX, METHOD Controls POSITIVE CONTROLS NEGATIVE CONTROLS Antibody Evaluation and Validation ANTIBODY SPECIFICATION SHEET PREDILUTED AND CONCENTRATED ANTIBODIES ANTIBODY VALIDATION STORAGE OF ANTIBODIES BLOCKING REACTIONS Validation Form for Antibodies and Tissue Controls MULTILINK BIOTINYLATED SECONDARY ANTISERA DAB REACTION PRODUCT INTENSIFICATION BUFFER SOLUTIONS Commonly Used Antibodies and Their Applications NEOPLASTIC TERMINOLOGY Quality Control RECOMMENDED QC FOR AN ANTIBODY POSITIVE AND NEGATIVE TISSUE CONTROLS RECOMMENDED QC FOR A TISSUE BLOCK DAILY QC OF IMMUNOHISTOCH EMISTRY STORAGE OF CONTROL SLIDES Standardization Troubleshooting Immunoperoxidase Techniques Staining Techniques BASIC PAP IMMUNOPEROXIDASE PROCEDURE ABC-IMMUNOPEROXIDASE PROCEDURE HRP ENZYME-LABELED POLYMER PROCEDURE References LEARNING ACTIVITIES Chapter 13: Enzyme Histochemistry Muscle Histology Pathologic Changes in Muscle Enzyme Histochemistry Oxidation and Reduction Properties of Enzymes Preservation of Enzymes Classification of Enzymes HYDROLASES OXIDOREDUCTASES TRANSFERASES Freezing Muscle Biopsy Specimens α-NAPHTHYL ACETATE ESTERASE STAIN FORMUSCLE BIOPSIES NAPHTHOL AS-D CHLOROACETATE ESTERASETECHNIQUE MAYER HEMATOXYLIN ATPASE STAIN ACID PHOSPHATASE IN MUSCLE BIOPSIES ALKALINE PHOSPHATASE STAIN FOR MUSCLEBIOPSIES NADH DIAPHORASE SUCCINIC DEHYDROGENASE (SDH) PHOSPHORYLASE STAIN FOR MUSCLE Nonenzymatic Procedures for Muscle Disorders MODIFIED GOMORI TRICHROME Acknowledgment References LEARNING ACTIVITIES Chapter 14: Electron Microscopy Fixation FIXATIVES FACTORS INFLUENCING FIXATION FIXATIVE SOLUTIONS Processing DEHYDRATION TRANSITIONAL SOLVENTS EMBEDDING MEDIA PROCEDURE FOR ROUTINE PROCESSING AND SPURREMBEDDING PROCEDURE FOR ROUTINE PROCESSING AND EPONEMBEDDING PROCEDURE FOR LR WHITE PROCESSING FORELECTRON MICROSCOPY IMMUNOLABELING Sectioning SECTION THICKNESS KNIVES CORRECTING PROBLEMS ENCOUNTERED INSECTIONING Staining Staining 0.5-μm Sections TOLUIDINE BLUE-BASIC FUCHSIN PROCEDURE TOLUIDINE BLUE STAINING Staining Thin Sections Special Techniques BLOOD CELL PREPARATION CELL SUSPENSIONS (FLUIDS, CULTURES, PARASITES,ETC) Processing Tissues Previously Embedded in Paraffin Processing Tissue from an H&E-Stained ParaffinSection Acknowledgment References LEARNING ACTIVITIES Chapter 15: Cytopreparatory Techniques Cytopreparation Collection GYNECOLOGIC CYTOLOGY NONGYNECOLOGIC CYTOLOGY Fixation PRE-FIXATIVES Smear Preparation DIRECT SMEARS FLUIDS MUCOID SPECIMENS SPARSELY CELLULAR SPECIMENS FINE NEEDLE ASPIRATIONS SPECIAL PROBLEMS CHOOSING THE BEST METHOD Liquid-Based Cytology Cell Blocks METHODS Cytology Staining HEMATOXYLIN OG-6 EA PAPANICOLAOU STAIN TOLUIDINE BLUE WET FILM CROSS CONTAMINATION SPECIAL STAINS References LEARNING ACTIVITIES Glossary Index