IN-SITU HYBRIDIZATION AT ELECTRON-MICROSCOPIC LEVEL

In the current study, we developed an improved method for ISH at the e lectron microscopic level, in which we employed pre-embedding hybridiz ation using a BrdU labeled probe followed by post-embedding immunoglob uline gold colloid staining (IGS). Because metabolic labeling of BrdU provides an almost evenly labeled DNA probe with high labeling index, pre-embeddingly hybridized BrdU-labeled probe preserves its detectable antigenicity even after the preparation of ultrathin sections. We dem onstrated precise cell identification, early stage of gene expression and alternative splicing patterns using this technique.