Spleen fixed in 4% PFA for 2h @ RT, washed 2x in PBS, then incubated ON in 30% Sucrose in PBS @ 4°C, froze away in OCT/Tissuetek. In general we use Tyramide Signal amplification for generate a brighter staining (e.g. PerkinElmer) but here not used. Detection with Streptavidin-Cy3 (Caltag, 1:500 in PBS for 30 min @ 20°C). The attatched picture shows a spleen section from a mice infected with 1x10exp6 L. monocytogenes for 24h. Nuclei were stained with DAPI shown in grey.
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Submitted Jul 17 2009
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