Simultaneous in situ hybridization using biotinylated and 35S-labeled riboprobes successfully detected two mRNAs in the same cell, though radiolabeled probe sensitivity was reduced by about 40%.
We used 35S-labeled and biotinylated cRNAs (riboprobes) to detect simultaneously two different mRNAs by in situ hybridization. In a first step we established the conditions under which each type of probe achieved the same high level of sensitivity. We then used these conditions to hybridize BHK cells infected with Theiler's virus, a murine picornavirus, with a mixture of a virus-specific biotinylated riboprobe and a 35S-labeled riboprobe specific for beta-actin mRNA. Both mRNAs could be detected in the same cell, although the sensitivity achieved by the radiolabeled probe was reduced by about 40% by the simultaneous hybridization with the biotinylated probe.
Ozden et al. (Sun,) conducted a other in Theiler's virus infection (in vitro). Simultaneous in situ hybridization with biotinylated and 35S-labeled riboprobes was evaluated on Simultaneous detection of two mRNAs. Simultaneous in situ hybridization using biotinylated and 35S-labeled riboprobes successfully detected two mRNAs in the same cell, though radiolabeled probe sensitivity was reduced by about 40%.