Course-based undergraduate research experiences (CUREs) using inquiry-based methodology provide a range of positive benefits to undergraduates and instructors. Yet, the required time and cost in designing and running CUREs with detailed data acquisition steps can lead to barriers in CURE implementation. This report describes an alternative approach to CUREs that utilizes free, open access 3D image volumes as data-rich resources for neurobiology CUREs. These open access image volume CUREs (ivCUREs) effectively combine the data acquisition and analysis steps within the course, allowing more time for students to critically evaluate their hypotheses and results, compare data with peers, and reflect on their experiences. Undergraduates in this 10-week ivCURE analyzed >670 excitatory synapses across two brain areas for the presence and origins of spinules within presynaptic boutons, and fully reconstructed 13 of these synapses in 3D. These data highlight the prevalence of these enigmatic synaptic features within excitatory presynaptic boutons, and their potential importance to neuronal function. Moreover, these results underscore key benefits to ivCURE implementation, including the (1) low-cost of experimental design and implementation, (2) ability to utilize the same data-rich image volume across multiple ivCUREs, (3) potential to generate publishable analyses, and (4) flexibility to scale projects and class sizes up at little to no cost. Opportunities for undergraduates to engage in inquiry-based ivCUREs that examine a host of unexplored questions in neurobiology will continue to grow, in parallel with rapid advances in 3D microscopy techniques and the increased availability and diversity of open access image volumes and analytical tools.