An optimized bioluminescent substrate for non-invasive imaging in the brain

Yichi Su; Joel R Walker; Mary P Hall; Mark A Klein; Xiang Wu; Lance P Encell; Kerriann M Casey; Lan Xiang Liu; Guosong Hong; Michael Z Lin; Thomas A Kirkland

doi:10.1038/s41589-023-01265-x

An optimized bioluminescent substrate for non-invasive imaging in the brain

Nat Chem Biol. 2023 Jun;19(6):731-739. doi: 10.1038/s41589-023-01265-x. Epub 2023 Feb 9.

Authors

Yichi Su^#¹, Joel R Walker^#², Mary P Hall³, Mark A Klein³, Xiang Wu^{4

5}, Lance P Encell³, Kerriann M Casey⁶, Lan Xiang Liu¹, Guosong Hong^{4

5}, Michael Z Lin^{7

8

9}, Thomas A Kirkland^{10

11}

Affiliations

¹ Department of Neurobiology, Stanford University, Stanford, CA, USA.
² Promega Biosciences, LLC, San Luis Obispo, CA, USA.
³ Promega Corporation, Madison, WI, USA.
⁴ Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA.
⁵ Wu Tsai Neurosciences Institute, Stanford University, Stanford, CA, USA.
⁶ Department of Comparative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
⁷ Department of Neurobiology, Stanford University, Stanford, CA, USA. mzlin@stanford.edu.
⁸ Department of Bioengineering, Stanford University, Stanford, CA, USA. mzlin@stanford.edu.
⁹ Department of Chemical and Systems Biology, Stanford University, Stanford, CA, USA. mzlin@stanford.edu.
¹⁰ Promega Biosciences, LLC, San Luis Obispo, CA, USA. thomas.kirkland@promega.com.
¹¹ Promega Corporation, Madison, WI, USA. thomas.kirkland@promega.com.

^# Contributed equally.

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.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
Brain / diagnostic imaging
Diagnostic Imaging*
Firefly Luciferin
Luciferins
Luminescent Measurements* / methods
Mice

Substances

Firefly Luciferin
Luciferins

Abstract

Publication types

MeSH terms

Substances

Grants and funding