Rm. Dickson et al., Simultaneous imaging of individual molecules aligned both parallel and perpendicular to the optic axis, PHYS REV L, 81(24), 1998, pp. 5322-5325
We report the first room temperature observation of an emission pattern fro
m a single fluorescent molecule signifying that the emission dipole is alig
ned along the optic (z) axis of a microscopic imaging system. This techniqu
e takes advantage of the x, y, and z-polarized evanescent fields generated
in total internal reflection and the usually nettlesome aberrations commonl
y encountered when imaging biological samples with high numerical aperture
oil-immersion objectives. For both z-oriented and transverse-oriented indiv
idual molecules of the carbocyanine dye DiIC(18) embedded in polymethyl-met
hacrylate (PMMA), calculated images accurately model the main features of o
bserved emission patterns.