Summary
Published in Sensors (2016), this peer-reviewed study used 3D Petri Dish® micro-molds to form scaffold-free 3D microtissues. Full citation: Chennell, George, et al. Imaging of Metabolic Status in 3D Cultures with an Improved AMPK FRET Biosensor for FLIM
Imaging of Metabolic Status in 3D Cultures with an Improved AMPK FRET Biosensor for FLIM
Research Overview
This study developed a way to map biochemical and physiological change inside 3D cultures non-invasively, using FRET biosensors expressed in tumor spheroids and read out by fluorescence lifetime imaging (FLIM).
The centerpiece is an improved AMP-activated protein kinase biosensor, T2AMPKAR, in which replacing the original ECFP donor with mTurquoise2 increased the dynamic range of the response to AMPK activation — demonstrated in 2D with the direct activator 991, then applied to 3D FLIM imaging of tumor spheroids to track metabolic status spatially and over time.
Key Discoveries
- Improved AMPK FRET biosensor (T2AMPKAR) increased dynamic range by swapping ECFP for mTurquoise2
- FLIM readouts enabled non-invasive metabolic mapping inside tumor spheroids
- Validated in 2D with the direct AMPK activator 991, then extended to 3D
