TY - JOUR
T1 - Total internal reflection fluorescence. Measurement of spatial and orientational distributions of fluorophores near planar dielectric interfaces
AU - Thompson, Nancy L.
AU - Burghardt, Thomas P.
N1 - Funding Information:
We thank Daniel Axelrod, Edward Hellen and Robert Fulbright of the University of Michigan for helpful discussions concerning fluorescence emission next to interfaces. This work was supported by National Institutes of Health grant GM37145 (N.T.) and by National Science Foundation Presidential Young Investigator Award DCB 8552986 (N-T.).
PY - 1986/11
Y1 - 1986/11
N2 - 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.
AB - 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.
KW - Evanescent wave
KW - Fluorescence microscopy
KW - Order parameter
KW - Ordered biological assembly
KW - Solid/liquid interface
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U2 - 10.1016/0301-4622(86)85069-4
DO - 10.1016/0301-4622(86)85069-4
M3 - Article
C2 - 3814748
AN - SCOPUS:0022804820
SN - 0301-4622
VL - 25
SP - 91
EP - 97
JO - Biophysical Chemistry
JF - Biophysical Chemistry
IS - 1
ER -