Note: Front Cover -- A Flexible System of Enzymatic Analysis -- Copyright Page -- Table of Contents -- PREFACE -- Part I: GENERAL PRINCIPLES -- Chapter 1. Pyridine Nucleotides -- Spectral Absorption and Fluorescence of Reduced Pyridine Nucleotides -- Limitations of Fluorometry -- Fluorescence Reference Standards -- Excitation and Emission Wavelengths and Light Filters -- Conversion of Pyridine Nucleotides in Alkali into Fluorescent Products -- Increased Fluorescence from DPNH and TPNH with Strong Alkali -- Stability and Selective Destruction of Reduced and Oxidized Forms of Pyridine Nucleotides -- Destruction of DPNH and TPNH in Acid and Stability in Alkali -- Destruction of DPN+ and TPN+ in Alkali and Stability in Acid -- Preparation, Standardization, and Storage of Pyridine Nucleotide Solutions -- Fluorometers -- Chapter 2. A Kinetic Primer for the Tissue Analyst -- NONENZYME KINETICS -- First-Order Reactions -- Second-Order Reactions -- ENZYME KINETICS -- One-Step Reactions with One Substrate -- First-Order Enzyme Reactions -- Mixed Zero-Order and First-Order Enzyme Reactions -- One-Step Reactions with Two Substrates -- Enzyme Reactions with Second-Order Kinetics -- Two-Step Reactions -- Other Factors Affecting Enzyme Kinetics -- Chapter 3. Glassware -- CONSTRICTION PIPETTES -- The Role of Surface Tension on Pipetting -- Effect of Changes in Surface Tension and Viscosity -- Pipetting Technique -- Special Considerations with Small Pipettes and Small Tubes -- Transferring the Whole Sample (" Total Transfer ") -- Common Pipetting Errors -- Rinsing Pipettes -- Making Constriction Pipettes -- Calibration of Pipettes -- Cleaning and Storage of Constriction Pipettes -- TUBES -- Fluorometer Tubes -- Tubes for 25-200 μl Volumes -- Tubes for Volumes of Less than 25 μl -- How to Make Micro Test Tubes -- Cleaning and Storage of Tubes. , Chapter 4. Typical Fluorometric Procedures for Metabolic Assays -- A ONE-STEP METABOLITE ASSAY WITH TPN+ -- Spectrophotometric Procedure -- Five Fluorometric Procedures for the Measurement of Glucose-6-P -- A ONE-STEP METABOLITE ASSAY WITH DPNH -- Spectrophotometric Procedure -- Four Fluorometric Procedures for the Measurement of α -ketoglutarate -- A MULTISTEP METABOLITE ASSAY (FOR INORGANIC PHOSPHATE) -- Spectrophotometric Procedure -- Four Fluorometric Procedures for the Measurement of Pi -- Chapter 5. Measurement of Enzyme Activities with Pyridine Nucleotides -- A ONE-STEP ENZYME ASSAY WITH TPN+ (ISOCITRIC DEHYDROGENASE) -- Spectrophotometric Procedure -- Three Fluorometric Procedures -- A ONE-STEP ENZYME ASSAY WITH DPNH (GLYCERO-P DEHYDROGENASE) -- Spectrophotometric Procedure -- Three Fluorometric Procedures -- A MULTISTEP ENZYME ASSAY (P-GLUCOMUTASE) -- Spectrophotometric Procedure -- Three Fluorometric Procedures -- Chapter 6. Improvement, Modification, Adaptation, Trouble Shooting, and Development of New Methods -- Simplification and Improvement -- Adaptation of Pyridine Nucleotide Methods to Increase Sensitivity -- Development of a New Method -- Chapter 7. Preparation of Tissues for Analysis -- Preparation for Enzyme Assays -- Preparation for Metabolite Analyses -- Preparation of Tissue Extracts -- The Use of Methanol-HCl in Preparing Extracts -- Fluorescence of Tissue Extracts -- Measurement of Pyridine Nucleotides in Tissues -- Chapter 8. Enzymatic Cycling -- TPN Cycle -- DPN Cycle -- ATP-ADP Cycle -- Glutamate- α-ketoglutarate Cycle -- Other Cycles -- Double Cycling -- Chapter 9. A Collection of Metabolite Assays -- ADP and AMP -- ALANINE -- ASPARTATE -- ATP and P-Creatine -- CITRATE -- Creatine -- Dihydroxyacetonephosphate: Method I (with Glycero-P-dehydrogenase) -- Fructose -- Fructose-6-phosphate. , Fructose-1, 6-diphosphate: Method I -- Fructose-1,6-diphosphate (Method II) Dihydroxyacetonephosphate (Method II), and Glyceraldehydephosphate -- Fumarate -- Glucose -- Glucose-1 -phosphate -- Glucose-6-phosphate -- Glucose-1,6-diphosphate -- Glutamate -- α-Glycerophosphate: Method I -- α-Glycerophosphate: Method II -- Glycogen -- Isocitrate -- Lactate: Method I -- Lactate: Method II -- Malate: Method I -- Malate: Method II -- Oxalacetate -- 6-P-Gluconate -- 3-P-Glycerate: Method I -- 3-P-Glycerate: Method II -- P-Pyruvate -- Pyruvate -- Total Nucleotide Triphosphates -- Uridine-5-diphosphoglucose -- Part II: QUANTITATIVE HISTOCHEMISTRY -- Introduction -- Chapter 10. Preparation of Tissues And Sections -- Freezing -- Storage of Frozen Tissues -- Mounting the Tissue for Cutting -- Sectioning the Tissue -- Drying of Samples -- Storage of Dry Sections -- Chapter 11. Dissection and Histological Control -- Dissection and Balance Room -- Removal of Sections from Evacuation Tube -- Dissection -- Handling Dissected Samples -- Identification and Histological Control -- Sample Carriers -- Defatting Frozen Dried Samples -- Sample Volume -- Chapter 12. The Quartz Fiber Fishpole Balance -- Measurement of Displacement and Useful Range -- Choice of Suitable Fiber. Sensitivity as a Function of Length and Diameter of the Fiber -- Balance Fibers -- Pans -- Balance Case -- Mounting the Fiber -- Lighting and Viewing -- Weighing -- Correction of Weight for Adsorption of Gases and Moisture -- Calibration of Balances and Testing Linearity -- Cleaning the Balance Fiber -- Chapter 13. Histochemical Analysis -- Introduction -- Enzyme Stability at High Dilution -- Addition of Sample and Reagent -- Oil Well Technique -- Reagent Wells -- Pipettes -- Microscope and Working Stage -- Addition of Sample and Reagent -- Special Considerations -- Part III: APPENDIX -- Appendix. , Acid-Base Made Easy -- Statistical Shortcuts -- Construction of Dissecting Knives -- Microtome Knife Sharpening -- Table of Indicators -- Preparation and Storage of Common Stock Solutions -- Normality o f Concentrated Acids and Bases -- Commercial Sources of Equipment and Supplies -- Abbreviated List of Atomic Weights -- References -- SUBJECT INDEX.