In vitro and in vivo molecular imaging techniques provide many unique opportunities to visualize and study biological processes. Bioluminescence is a ubiquitous and commonly used imaging modality for visualizing biological processes in vivo. Bioluminescence is the emission of light from biochemical reactions that occur within a living organism. This technique employs visible light and interfaces readily with most cell and tissue types, making it a versatile technology for preclinical studies and for the early detection of disease progression. Luciferin, a substrate of luciferase that plays a key role in the light emitted from fireflies, is at the heart of bioluminescence imaging and produces noticeable light outputs. Despite the great advancement in bioluminescence techniques, current methods for luciferin synthesis are low yielding and complex. The present technology relates to bioluminescence and in particular to methods and systems for the synthesis of D-aminoluciferin precursor and related compounds.