David Mason, Julia Jones
In situ hybridisation (ISH) is an invaluable experimental technique when considering the holistic process of transcription through to translation. Without good tools to identify sequence-based structures within cells and tissues, researchers are left with only half of the story when looking at protein expression. While there are many experimental approaches, including a rainbow of chromogens and a choice of brightfield and fluorescent approaches, the Image Analysis of ISH-based experiments remains a significant block to many looking to explore these datasets. This chapter aims to highlight some of the considerations when approaching the image analysis of ISH experiments, as well as showing a step-by-step walkthrough for analysis using tools developed by Visiopharm, an industry leader in digital pathology image analysis with over 20 years of experience in the field.