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Principles and Applications of Fluorescence Spectroscopy

Principles and Applications of Fluorescence Spectroscopy

Jihad Rene Albani

ISBN: 978-0-470-69133-5

Apr 2008, Wiley-Blackwell

264 pages



Fluorescence spectroscopy is an important investigational tool in many areas of analytical science, due to its extremely high sensitivity and selectivity. With many uses across a broad range of chemical, biochemical and medical research, it has become an essential investigational technique allowing detailed, real-time observation of the structure and dynamics of intact biological systems with extremely high resolution. It is particularly heavily used in the pharmaceutical industry where it has almost completely replaced radiochemical labelling.

Principles and Applications of Fluorescence Spectroscopy gives the student and new user the essential information to help them to understand and use the technique confidently in their research. By integrating the treatment of absorption and fluorescence, the student is shown how fluorescence phenomena arise and how these can be used to probe a range of analytical problems. A key element of the book is the inclusion of practical laboratory experiments that illustrate the fundamental points and applications of the technique.

1. Absorption spectroscopy theory.

2. Determination of Calcofluor White Molar extinction coefficient value in absence and presence of α1-acid glycoprotein.

3. Determination of kinetic parameters of Lactate dehydrogenase (LDH).

4. Hydrolysis of p-nitropenyl-β-D-galactoside with β-galactosidase from E.coli.

5. Starch hydrolysis by amylase.

6. Detrermination of the pK of a dye.

7. Fluorescence spectroscopy principles.

8. Effect of the structure and the environment of a fluorophore on its absorption and fluorescence spectra.

9. Fluorophores characterization and importance in biology.

10. Fluorescence quenching.

11. Fluorescence polarization.

12. Interaction between ethidium bromide and DNA.

13. Lens culinaris agglutinin: dynamics and binding studies.

14. Förster energy transfer.

15. Binding of TNS on bovine serum albumin at pH 3 and pH 7.

16. Comet Test for environmental genotoxicity evaluation: a fluorescence microscopy application.

17. Questions and exercises

  • Straightforward overview of absorption and fluorescence shows the student how the phenomenon arises and how it can be used in the course of their research.
  • Highly practical approach shows non-specialists how to use the technique to investigate chemical and biochemical problems and generate sophisticated results.
  • Contains many easy to perform laboratory experiments to reinforce the clear explanations.