In situ hybridization (both radioactive and nonradioactive) and spatiotemporal gene expression analysis

Section in situ hybridization using either radioactive or nonradioactive labeled cDNA probes is an invaluable technique that enables the investigator to detect and localize mRNA expression within tissue sections and cells. Here, we describe the labeling of (35)S-UTP radioactive and nonradioactive digoxigenin probes, preparation of tissue sections, hybridization, and washing of non-hybridized probes, followed by the detection of radioactive signals via dipping in nuclear emulsion and the immunohistochemical and subsequent colorimetric detection of nonradioactive signals.