Image analysis has become a central tool in modern biology. While the human eye analyze images, its assessments are often qualitative. Computers provide quantitative, unbiased measurements, and enable the automation of the analysis, leading to a larger number of processed samples and a greater power of downstream statistical tests. In this course, we will discuss the main steps in the analysis of digital images, with an emphasis on different modalities of microscopy data, including confocal, TIRF, and super-resolution. Topics will include image display, filtering, segmentation, mathematical morphology, and measurements. Lectures will be complemented with examples from the current literature. Students will also have the opportunity to develop solutions to the analysis of images from their own research in a final project.