Sensitive fluorescence in situ hybridization on semithin sections of adult Schistosoma mansoni using DIG-labeled RNA probes
2020
Abstract
In situ hybridization is a tool for evaluation of gene expression within tissues or single cells. This protocol describes optimized sensitive fluorescence detection of gene transcripts (mRNAs) in semithin sections of Schistosoma mansoni adult worms using specifically designed and labeled RNA probes.
Due to improved methodologies in tissue preservation, sectioning, amplification of fluorescent signal, and prehybridization tissue treatment, it is possible to detect transcripts in the fine structures of schistosomes. The protocol is sensitive enough to detect very low abundance targets.
This procedure is optimized for tissues derived from S. mansoni adult worms; however, it can be successfully applied to other trematode species.