Use of image analysis software as a tool to visualize non-radioactive signals in plant in Situ analysis

In situ hybridization (ish) allows the visualization of gene expression in tissues at high microscopic resolution. Interference by plant tissue pigments generally confers higher sensitivity to radioactive ish, relative to non-radioactive ish using hapten labeled probes. The increased resolution is partially due to image acquisition methods in radioactive ish experiments. However, radioactive ish has many drawbacks including short probe life, safety concerns associated with the use of radioactive materials, and slow development of signal. In this report, we show how commercially available image analysis software can be used to extract data from non-radioactive ish images to gain a substantial increase in resolution. We provide a comparison between detecting a probe (CELLULOSE SYNTHASE) that is expected to produce a consistent, detectable signal in all growing tissues with detection of a probe (LEAFY) that is expected to produce a signal only in specific tissues.