Project Summary Cryogenic electron microscopy (cryo-EM) is a rapidly emerging technology for structural biology. While excitement and demand for the technology are high, the pace of training cryo-EM practitioners lags far behind. There are two major hurdles to the development of effective training programs: (1) potential learners come from a wide range of backgrounds and career stages, from graduate students to established structural biologists; and (2) learning cryo-EM requires many hours of hands-on “behind the wheel” training on specialized, expensive equipment that is often unavailable to learners at their home institutions. While some hands-on schools exist for cryoEM, they train only a handful of students a year, while demand for the technique requires orders of magnitude more. Workshops enroll more students, but are too short to teach all the necessary theory and lack a hands-on component. To train a skilled workforce of cryo-EM practitioners, here I propose to expand upon my successful “Getting Started in Cryo-EM” video lectures and create a comprehensive online curriculum covering the theory and practice of all major cryo-EM modalities including cryo-EM single particle reconstruction (SPR), electron cryotomography (cryo-ET) and microcrystal electron diffraction (micro-ED). This curriculum will include many tens of hours of new videos and also the supporting materials needed to maximize the usefulness of these videos. More specifically, my colleagues and I will produce new videos teaching the theory and practice of cryo-ET (including special sample prep issues like correlated light and electron microscopy and cryo-FIB milling), micro-ED, and facility management/advanced topics. We will also produce videos that introduce basic principles of cryo-EM image processing and prepare students to move from our materials to the specialized tutorials already available for several popular image processing packages. We will also create supporting materials including homework problems, open-ended and multiple-choice exam questions, training manuals with specific standardized practice exercises, and plans for training programs. Together with staff from the national cryo-EM service centers and my fellow curriculum development program awardees, I will work to develop a “merit badge” system to certify cryo-EM practitioners on specific skills and instruments. Materials will be designed in modules that can be mixed and matched to meet the specific needs of the learner. In conjunction with supervised training at a user’s home facility or national cryo-EM service center, the curriculum will offer everything a student needs to understand the theory of, and begin practicing, their desired cryo-EM technique.