Research Papers

Fluorescence emission patterns near glass and metal-coated surfaces investigated with back focal plane imaging

[+] Author Affiliations
Alexa L. Mattheyses

University of Michigan, Biophysics Research Division, Ann Arbor, Michigan 48109

Daniel Axelrod

University of Michigan, Biophysics Research Division, Ann Arbor, Michigan 48109 and University of Michigan, Department of Physics, Ann Arbor, Michigan 48109

J. Biomed. Opt. 10(5), 054007 (September 19, 2005). doi:10.1117/1.2052867
History: Received January 13, 2005; Revised April 25, 2005; Accepted April 29, 2005; Published September 19, 2005
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Often fluorophores observed by microscopy are located close to a planar surface as in total internal reflection microscopy or single molecule studies. The optical properties of fluorescence excitation and emission near a planar surface (possibly metal film coated) between two dielectrics are well understood from the viewpoint of classical electromagnetic theory. We present an experimental method whereby the angular dependence of the emission pattern of a fluorophore near a bare surface or a metal film coated surface that supports surface plasmon resonance can be measured with microscope optics. The technique involves an alteration of the microscope optics to directly record (on a digital CCD camera) the intensity pattern at the objective’s back focal plane, which directly maps the angular emission pattern of fluorescence. The experimental emission profile on both glass and aluminum-coated surfaces is anisotropic with a peak at either the critical angle or both the critical angle and the surface plasmon angle. The observed profiles agree well with computer calculations and suggest some optical modifications that are potentially useful in cell biophysics.

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© 2005 Society of Photo-Optical Instrumentation Engineers

Citation

Alexa L. Mattheyses and Daniel Axelrod
"Fluorescence emission patterns near glass and metal-coated surfaces investigated with back focal plane imaging", J. Biomed. Opt. 10(5), 054007 (September 19, 2005). ; http://dx.doi.org/10.1117/1.2052867


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