Suggested beads for STED PSF acquisition and parameter estimation

Recording Beads is a necessary first step to distill an Experimental Psf.
The big advantage of imaging STED beads, and extracting a PSF from that using the PSF distiller, is that the specific STED microscopic parameters are estimated. These parameters can subsequently be used for deconvolution of the STED image of your specific sample. See also STED deconvolution.

For STED microscopy, the following fluorescent beads were used with good results:
FluoSpheres® Carboxylate-Modified Microspheres 0.040 micron (Yellow-Green Fluorescent (em/ex: 505/515 nm); Catalog Number F-8795 ) from Life Technologies - Molecular Probes.