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Fig. 3. In situ hybridization of TRITC-labelled PNA probes with amplified nucleoli. Probes antisense to Xenopus SRP RNA (A-D) and sense of yeast SRP RNA (control, E-G) were hybridized to GV spread preparations. Phase-contrast images (A,E) show nucleoli (No), chromosomes (Ch) and Cajal bodies (CB). DAPI images (B,F) show staining of fibrillar centres and chromomeres. TRITC fluorescence shows that the antisense probe hybridizes to nucleoli, although not significantly to the fibrillar centres (C). This differential localization is seen clearly in merged images in which the DAPI image is computer-coloured green (D). Coincidence of signals appears as yellow: the small amount of coincidence in nucleoli can be accounted for by overlapping nucleolar components, whereas complete coincidence is seen with the wide-spectrum autofluorescence of a contaminating yolk platelet (YP). The control probe fails to hybridize to nucleoli, although signal is seen over the yolk platelets (YP) in (G).





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