Mouse 3T3 fibroblast immunostained for the nucleolar protein nucleostemin (left, green) and fibrillarin (right, red) marking nucleolar fibrillar centers.
Data were collected using a Leica DMIRB microscope equipped with a 100x objective (N.A. 1.4) and appropriate filter sets, and images captured using a Quantix 57 CCD camera (Roper Scientific Photometrics). For high resolution spatial mapping, three-dimensional optical stacks (containing 21 consecutive 0.25 micron slices) were captured using a PIFOC microscope focusing drive (Polytec PI). Images were dark current subtracted, intensity scaled, and in some cases, z-stacks deconvolved using exhaustive photon reassignment. Shown is a midplane of a deconvolved stack. See Fig 2EF in Politz et al., 2005 Mol Biol Cell 16:3401-3410.
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