Nanometer axial resolution by three-dimensional supercritical angle fluorescence microscopy.
We report a noninvasive fluorescence microscopy method and demonstrate nanometer resolution along the optical axis. The technique is based on the influence of the microscope slide on the angular intensity distribution of fluorescence. Axial positions are determined by measuring the proportion of light emitted below the critical angle of total internal reflection, which behaves in a classical way, and light emitted above the critical angle, which is exponentially dependent on the distance of the fluorophore from the microscope slide.
Winterflood
Christian M
CM
Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.
Ruckstuhl
Thomas
T
Verdes
Dorinel
D
Seeger
Stefan
S
D000818:Animals; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008863:Microspheres; D008870:Microtubules; D041681:NIH 3T3 Cells; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods*; D010316:Particle Size