Article Synopsis
- Bioluminescence imaging (BLI) is a crucial technique for visualizing cells and biochemical activity in living organisms, but its application in the brain has been limited due to poor performance of existing luciferase substrates.
- The new substrate, cephalofurimazine (CFz), significantly enhances imaging sensitivity by producing over 20 times more signal in the brain when used with Antares luciferase compared to standard substrates.
- This advancement enables detailed, real-time imaging of brain activity and calcium levels in specific neuron types in freely moving mice, expanding the potential for NanoLuc-based research in neuroscience.
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Article Abstract
Bioluminescence imaging (BLI) allows non-invasive visualization of cells and biochemical events in vivo and thus has become an indispensable technique in biomedical research. However, BLI in the central nervous system remains challenging because luciferases show relatively poor performance in the brain with existing substrates. Here, we report the discovery of a NanoLuc substrate with improved brain performance, cephalofurimazine (CFz). CFz paired with Antares luciferase produces greater than 20-fold more signal from the brain than the standard combination of D-luciferin with firefly luciferase. At standard doses, Antares-CFz matches AkaLuc-AkaLumine/TokeOni in brightness, while occasional higher dosing of CFz can be performed to obtain threefold more signal. CFz should allow the growing number of NanoLuc-based indicators to be applied to the brain with high sensitivity. Using CFz, we achieve video-rate non-invasive imaging of Antares in brains of freely moving mice and demonstrate non-invasive calcium imaging of sensory-evoked activity in genetically defined neurons.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10229426 | PMC |
| http://dx.doi.org/10.1038/s41589-023-01265-x | DOI Listing |
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