Variable incidence angle linear dichroism (VALiD): A technique for unique 3D orientation measurement of fluorescent ensembles

Academic Article


  • A fundamental challenge with fluorophore orientation measurement is degeneracy, which is the inability to distinguish between multiple unique fluorophore orientations. Techniques exist for the non-degenerate measurement of the orientations of single, static fluorophores. However, such techniques are unsuitable for densely labeled and/or dynamic samples common to biological research. Accordingly, a rapid, widefield microscopy technique that can measure orientation parameters for ensembles of fluorophores in a non-degenerate manner is desirable. We propose that exciting samples with polarized light and multiple incidence angles could enable such a technique. We use Monte Carlo simulations to validate this approach for specific axially symmetric ensembles of fluorophores and obtain optimal experimental parameters for its future implementation.
  • Published In

  • Optics Express  Journal
  • Digital Object Identifier (doi)

    Author List

  • Blanchard AT; Brockman JM; Salaita K; Mattheyses AL
  • Start Page

  • 10039
  • End Page

  • 10061
  • Volume

  • 28
  • Issue

  • 7