Artificial intelligence (AI) is more than just a buzzword—it is now a permanent fixture in everyday life and in scientific research. AI is used in all forms of research, from diagnostics and drug discovery to imaging software. Even the microscope—a mainstay of most research laboratories—has also become self-learning. Given that the mainstream applications for new AI methods are still building momentum, it may not be clear to researchers the types of scenarios that are well suited to AI-based solutions. Our goal in this supplement is to help inform readers of the possibilities for their own research presented by deep learning–based imaging and analysis methods for microscopy.