Total internal reflection fluorescence. Measurement of spatial and orientational distributions of fluorophores near planar dielectric interfaces

Nancy L. Thompson, Thomas P. Burghardt

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

The fluorescence collected from a fluorophore which is near a planar interface and is excited by a laser beam that is totally internally reflected at the interface depends on the direction of the absorption and emission transition dipole moments of the fluorophore with respect to the interface, on the distance from the fluorophore to the interface, on the angle of incidence and polarization direction of the exciting beam, and on properties of the collection optics. Expressions are derived for the excitation and subsequent emission and collection of fluorescence from a population of fluorophores near a planar interface. Presented is a general model-independent method of obtaining characteristic parameters of the spatial and orientational distribution of the population of fluorophores, from a measure of the fluorescence collected as a function of the polarization and the incidence angle of the totally internally reflected laser beam. The method is illustrated with several simulation calculations.

Original languageEnglish (US)
Pages (from-to)91-97
Number of pages7
JournalBiophysical Chemistry
Volume25
Issue number1
DOIs
StatePublished - Nov 1986

Keywords

  • Evanescent wave
  • Fluorescence microscopy
  • Order parameter
  • Ordered biological assembly
  • Solid/liquid interface

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Organic Chemistry

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