Abstract
Since its discovery in fireflies, bioluminescence has been used in a wide range of biological assays, from reporter gene assays to in vivo imaging. Bioluminescent light is produced from the enzymatic action of luciferase on its substrate luciferin using ATP as a cofactor. Recently, subcellular targeting of luciferase to neural synaptic vesicles has led to the development of the sensor Syn-ATP that allows for quantitative measurement of presynaptic ATP levels. Manual analysis of presynaptic bioluminescence signals from Syn-ATP is challenging due to signal heterogeneity and cellular motion in long imaging sessions. Here, we present a pipeline for semi-automated image analysis of Syn-ATP signals in the nerve terminals of hippocampal neurons. Our method streamlines data analysis and reduces user-introduced bias, thus enhancing the reproducibility and reliability of quantitative ATP imaging.
Competing Interest Statement
The authors have declared no competing interest.