The Mesoscale to Molecules Bioimaging Technology Program, within the Department of Energy?s (DOE) Office of Biological and Environmental Research (BER), sponsors research that addresses grand challenges for biology. These grand challenges are centered around developing a better understanding of the spatial and temporal distributions of key metabolites and biomarkers associated with microorganisms and plants related to bioenergy and environmental systems. The Bioimaging Technology Program is advancing new and improved instruments that are being designed and constructed from the ground up, adapted from other scientific disciplines, or upgraded with new capabilities to create a comprehensive and versatile toolbox for imaging biological dynamics and chemistry. These instrument development approaches involve (1) hypothesis-based technologies focused on solving particular challenges for plant biology, biofuels, and biogeochemistry and (2) broad-based systems and design-based technologies that can lead to new discoveries. Both approaches are being pursued with versatility, transferability, and portability in mind to foster widespread adoption by the larger science community for future field and laboratory studies.