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Future research into the prognostic capabilities of the Ki67/MART1 double stain will be simplified by the novel opportunity to quantify MART1-verified Ki67 indices automatically.
By using Leica ultra-cut UCT we obtained semi-thin sections which were stained with toulidine blue and ultra- thin section (60 nm) were obtained and double stained with uranyl acetate and lead citrate for 10 min.
When sections were double stained with F4/80 antibody labeled with FITC and anti-CD166 labeled with phycoerythrin, colocalization could be identified easily as yellow fluorescent signals (Figure 4, C) with 1% of SBB.
Ultrathin sections were double stained with uranyl acetate--lead citrate and examined with a Philips CM100 transmission electron microscope (Philips Electron Optics, Eindhoven, The Netherlands).
The double stains were performed on the slides that were previously stained with pan-CK or CAM5.2 using diaminobenzidine as the chromogen.