Principles and Applications of Fluorescence Spectroscopy

  • ID: 2178617
  • Book
  • 264 Pages
  • John Wiley and Sons Ltd
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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.

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1. Absorption spectroscopy theory.

2. Determination of Calcofluor White Molar extinction coefficient value in absence and presence of 1–acid glycoprotein.

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

4. Hydrolysis of p–nitropenyl– –D–galactoside with –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

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Jihad–René Albani is Associate Professor and head of the Laboratoire de Biophysique Moléculaire at the Université des Sciences et Technologies de Lille, France
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