Immunofluorescence stimulated emission depletion microscopy.
We report immunofluorescence imaging with a spatial resolution well beyond the diffraction limit. An axial resolution of approximately 50 nm, corresponding to 1/16 of the irradiation wavelength of 793 nm, is achieved by stimulated emission depletion through opposing lenses. We have demonstrated not only that an antibody-tagged label is stable enough to be recorded in this microscopy mode, but also that subdiffraction resolution can be obtained using a standard immunofluorescence preparation.
Dyba
Marcus
M
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37070 Göttingen, Germany.
Jakobs
Stefan
S
Hell
Stefan W
SW
D002478:Cells, Cultured; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000451:nursing*; D008870:Microtubules / Q000648:ultrastructure*; D012680:Sensitivity and Specificity