ViewRNA Tissue In situ Hybridization (ISH)
The MPC held a workshop on Feb. 22-23 with Mike Shure, PhD, field application scientist Affymetrix-Panomics Solutions, to validate an insulin ISH probe on normal human pancreas. Assay validation consisted of pretreatment optimization for tissue heating and protease digestion times. Insulin mRNA is shown in the attached image from one of several combinations that yielded outstanding results. These ISH probes are reported to be capable of detecting single RNA molecules through multiple pairs of specific probe pairs and multi-amplification of the primary signal. Fast Red substrate allows either brightfield or fluorescent signal localization. More information can be found on the Affymetrix website including their online catalogue of several hundred probes and information for custom probe construction. Dr. Ann Fu is available for assay consultations and can now offer this service to Molecular Pathology Core users. This technology is particularly valuable when suitable antibodies are not available and when antibody localization or changes in protein expression needs verification through an alternate method.