Fluorescence Assay in Biology and Medicine - 1st Edition - ISBN: 9780127058023, 9781483258812

Fluorescence Assay in Biology and Medicine

1st Edition

Authors: Sidney Udenfriend
Editors: Bernard Horecker Nathan O. Kaplan Julius Marmur
eBook ISBN: 9781483258812
Imprint: Academic Press
Published Date: 1st January 1969
Page Count: 672
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Molecular Biology: An International Series of Monographs and Textbooks: Fluorescence Assay in Biology and Medicine, Volume II covers the many applications of fluorescence and phosphorescence.

This book discusses the principles of fluorescence polarization, comparison of luminescence methods of analysis, and direct measurement of fluorescence decay times. The photodecomposition, sulfhydryl compounds, determination of primary structure, and fluorescent staining are also deliberated. This text likewise covers the assay of purines in nucleic acid hydrolyzates, formyltetrahydrofolate synthetase, and ovarian hormones.

This volume is valuable to chemists, physicists, and biophysicists intending to use fluorescence in studying reaction mechanisms and elucidate the structure of complex biopolymers.

Table of Contents


Chapter 1: Principles of Fluorescence

I. General Remarks

II. Structure and Fluorescence

III. Delayed Fluorescence

IV. Nonradiative Energy Transfer

V. Quantum Yield and Lifetime of Fluorescence

VI. Lifetime ofthe Excited State

VII. Fluorescence Polarization


Chapter 2: Phosphorimetry as a Means of Chemical Analysis

I. General Remarks

II. Comparison of Limiinescence Methods of Analysis

III. Instrumentation and Methodology in Phosphorimetry

IV. Applications of Phosphorimetry


Chapter 3: Instrumentation

I. Introductory Remarks

II. Visual Detection

III. Instrumental Detection

IV. Filter Fluorometers

V. Fluorescence Spectrometry

VI. Microspectrofluorometers

VII. Direct Measurement of Fluorescence Decay Times

VIII. Quantum Yield

IX. Fluorescence Polarization

X. Chromatograms, Electrophoretograms, and Solids

XI. Automation

XII. Rapid Reactions


Chapter 4: Practical Considerations

I. General Remarks

II. Cuvettes and Solvents

ΙII. Problemsof Light Absorption

IV. Light Scattering

V. Photodecomposition

VI. Instrumental Standardization and Calibration

VII. Limits of Sensitivity


Chapter 5: Amino Acids, Amines, and Their Metabolites

I. General Methods

II. Aromatic Amino Acids, General

III. Phenylalanine and Metabolites

IV. Tyrosine and Metabolites

V. Tryptophan and Metabolites

VI. Histidine and Histamine

VII. Arginine

VIII. Lysine

IX. Sulfhydryl Compounds

X. Acetylcholine

XI. Urea

XII. Spermine and Spermidine


Chapter 6: Proteins and Peptides

I. General Remarks

II. Quantitative Assay

III. Determination of Primary Structure

IV. Structure and Conformation

V. Specific Proteins


Chapter 7: Immunology

I. General Remarks

II. Fluorescent Staining

III. Chemical Studies of Antigens and Antibodies


Chapter 8: Vitamins, Coenzymes, and Their Metabolites

I. General Remarks

II. Vitamin A

III. Thiamine and Cocarboxylase

IV. Riboflavin and Flavin Coenzymes

V. Pyridine Nucleotides, Nicotinamide, and Their Metabolites

VI. Pyridoxine and Its Congeners

VII. Ascorbic Acid

VIII. Vitamin D

IX. Pteridines

X. Vitamin B12

XI. Pantothenic Acid

XII. Ubiquinone


Chapter 9: Carbohydrates, Aldehydes, Ketones, and Lipids

I. Carbohydrates

II. Keto Acids and Krebs Cycle Compounds

III. Alcohols, Aldehydes, and Ketones

IV. Lipids


Chapter 10: Nucleic Acids, Nucleotides, Purines, and Pyrimidines

I. General Remarks

II. Pyrimidines and Their Nucleosides and Nucleotides

III. Assay of Purines in Nucleic Acid Hydrolyzates

IV. Nucleic Acids

V. Nucleic Acid Assay


Chapter 11: Porphyrins and Their Metabolites

I. General Remarks

II. Assay Procedures

IIΙ. Theoretical Considerations


Chapter 12: Fluorometric Assay of Enzyme Activity

I. General Remarks

II. Hydrolytic Enzymes

III. Oxidative Enzymes

IV. Transaminases

V. Dehydrogenases

VI. Phosphoglucoisomerase

VII. Creatine Kinase

VIII. Formyltetrahydrofolate Synthetase

IX. Aromatic L-Amino Acid Decarboxylase

X. Chemiluminescence


Chapter 13: Fluorescence as a Probe into Enzyme Mechanisms

I. General Remarks

II. Pyridine Nucleotide Enzymes

III. Flavin Enzymes

IV. Vitamin B6 Enzymes

V. Proteases

VI. Aryl Hydroxylase

VII. Lysozyme

VIII. Uridine Diphosphogalactose 4-Epimerase

IX. Muramidase

X. Interaction ofMyosin and ATP

XI. Tyrosinase

XII. Nucleases

XIII. Carbonic Anhydrase


Chapter 14: Steroids

I. General Remarks

II. Cholesterol

III. Ovarian Hormones

IV. Testosterone

V. Adrenal Steroids

VI. Bile Acids

VII. Steroid Drugs


Chapter 15: Plants

I. Chlorophyll and Photosynthesis

II. Alkaloids and Other Compounds in Plants


Chapter 16: Inorganic Constituents

I. General Remarks

II. Magnesium

III. Calcium

IV. Zinc

V. Aluminum

VI. Selenium

VII. Uranium

VIII. Fluoride

IX. Cyanide

X. Ammonia

XI. Hydrogen Peroxide

XII. Ozone


Chapter 17: Drugs and Toxic Agents

I. General Remarks

II. Chemotherapy of Infectious Diseases

III. Carcinostatic Agents

IV. Analgesics

V. Sedatives and Tranquilizers

VI. Antidepressants

VII. Hallucinogenic Agents

VIII. Parasympathetic Agents

IX. Antihistamines

X. Drugs Affecting the Cardiovascular System


Chapter 18: Fluorescence as a Probe into Mechanisms in Intact Cells and Subcellular Elements

I. General Remarks

II. Membranes and Transport Mechanisms

III. Oxidation-Reduction Mechanisms in Intact Cells and Subcellular Elements

IV. Measurement of Enzymes in Cells

V. Proteins

VI. Nucleic Acids and Chromosomes

VII. Localization and Identification of Substances in Tissue


Chapter 19: Public Health and Law Enforcement

I. General Remarks

II. Air Pollution

III. Food Inspection and Analysis

IV. Sanitation

V. Medical Diagnosis

VI. Forensic Medicine, Chemistry, and Criminology


Appendix I: Proposal for Standardization of Methods of Reporting Fluorescence Emission Spectra Text

Appendix II: Subcommittee on Fluorescence Spectroscopy

Method of Test for Resolution and Wavelength Accuracy

Method of Test for Linearity of Fluorescence Meters

Method of Test for Minimum Detectable by Fluorescence Spectrometers—Quinine Sulfate

Method of Test for Minimum Detectable by Fluorescence Spectrometers—Cyclohexane

Appendix ΙII: Practical Aspects of the Calibration of a Fluorescence Spectrometer



Author Index

Subject Index


No. of pages:
© Academic Press 1969
Academic Press
eBook ISBN:

About the Author

Sidney Udenfriend

About the Editor

Bernard Horecker

Nathan O. Kaplan

Julius Marmur

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