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Upgrade of a Scanning Confocal Microscope to a Single-Beam Path STED Microscope

Fig 3

Diffraction-unlimited imaging of crimson beads.

(A) Confocal and STED image of the same preparation of 20 nm crimson beads on a glass substrate, embedded in Mowiol/DABCO. STED power 14.5 mW (at the objective’s back aperture) at a repetition rate of 2.5 MHz. (B) Profile plots along the dashed white line in (A). (C) Full width at half maximum (FWHM) values determined by fitting 2D-Gaussian single peaks to STED images taken at different STED powers. Each data point represents the mean FWHM value of 10–20 peaks with the corresponding standard deviation. The solid lines show the result of a simulation with a function proportional to one over the square root of the STED intensity (Eq 3).

Fig 3

doi: https://doi.org/10.1371/journal.pone.0130717.g003