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Bioluminescence Imaging

NanoLuc® Luciferase allows sensitive, bright bioluminescent imaging with low background and no excitation required.

Bioluminescent imaging of cells and molecular processes in whole animals provides important insights when studying normal physiology, monitoring disease progress or understanding response to therapy. NanoLuc® reporter technologies provide new tools for studying biological processes within whole animal models. These bright and small reporter options allow for versatile in vivo applications ranging from quantifying changes in tumor growth to visualizing viral replication and spread using engineered NanoLuc® reporter viruses. 

Whole Animal Imaging

NanoLuc® reporters provide high sensitivity and low background when imaged in superficial tissues and have also been used successfully to image events in deeper tissues. ATP-independence of these reporters allows for in vivo monitoring of both intracellular and extracellular events.

In addition, several in vivo imaging strategies have been developed using NanoLuc®-based BRET reporters. These techniques utilize the bright NanoLuc® signal to excite red-shifted fluorescent acceptor proteins, creating enhanced deep tissue imaging solutions.

The Nano-Glo® In Vivo Substrate, fluorofurimazine (FFz),  is an optimized reagent designed specifically for in vivo detection of NanoLuc® Luciferase, NanoLuc® fusion proteins, or reconstituted NanoBiT® Luciferase. This aqueous soluble reagent provides increased substrate bioavailability in vivo, leading to bright signals, and has handling requirements compatible with in vivo workflows. In addition, substrate specificity allows NanoLuc® and firefly luciferases to be used together for dual luciferase molecular imaging studies, providing even more options for creating whole animal reporter models.

Nano-Glo® In Vivo Substrate, fluorofurimazine (FFz) is available as early access material. Please enquire to order product or request information.

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Imaging luciferase activity in live mice.
Nanoluc in vivo imaging

Learn more about imaging with NanoLuc® Luciferase in these resources:

Read about the development of NanoLuc® substrates in the paper Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals (Su, Y. et al. (2020)  Nature Methods 17, 852–860.

Read more about in vivo applications of NanoLuc® technology in the blog NanoLuc® Luciferase: Brighter Days Ahead for In Vivo Imaging.

Bioluminescent Imaging in Live Cells

In the cell, things are very dynamic; they are happening every second. The bioluminescence live-cell imaging was what allowed us to discover this dynamic"  
Dr. Li-Fang Chu, Assistant Professor, University of Calgary

Researchers in Dr. Chu's lab observed the exact timing of oscillating genes in early human development for the first time using bioluminescence live-cell imaging.  You can read more about this work in the blog: Observing the human developmental clock with bioluminescence live-cell imaging.



 Control of subcellular localization is an important mechanism for regulating the function and signaling activity of many proteins. For example, protein translocation from the cytosol to the nucleus or protein recruitment to the plasma membrane can be key events in signaling pathway activation. Bioluminescent imaging (BLI) can be used to monitor subcellular protein localization, allowing direct visualization of protein dynamics in living cells without the need for repeated sample excitation.

The NanoLuc® reporter is well-suited for use as a protein tag in BLI studies. The extreme brightness means that exposure times can be reduced to only a few seconds, compared to the minutes required for other luminescent reporter proteins. Also, the small size of NanoLuc® makes it less likely to perturb the normal biology or functionality of the fusion partner. New Nano-Glo® Extended Live Cell Substrates provide options for long-term imaging studies.

Monitoring translocation of a NanoLuc® Fusion Protein using Bioluminescence Imaging (BLI). HEK293 cells expressing a Protein Kinase C (PKC)-NanoLuc luciferase fusion protein were measured for 20 minutes following PMA treatment and application of furimazine substrate. BLI was performed on an Olympus® LV200 Bioluminescence Imager.