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 versatile in vivo applications ranging from quantifying changes in tumor growth to visualizing viral replication and spread using engineered NanoLuc® reporter viruses.

In Vivo Bioluminescence 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 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® Fluorofurimazine In Vivo Substrate (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.

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8060ca — Imaging luciferase activity in live mice.

Image the Brain of Living Animals with NanoLuc® Luciferase

Nano-Glo® Cephalofurimazine (CFz9) In Vivo Substrate enables the detection of NanoLuc® Luciferase in the brain of living animals. CFz9 was designed to cross the blood-brain barrier, enabling noninvasive exploration of central nervous system physiology and anatomy and studies of therapeutic distribution to the brain.

Request early access to CFz9 In Vivo Substrate.

Learn more about the CFz9 substrate in this webinar.

Learn more about imaging with NanoLuc® Luciferase

Webinar

Advancing In Vivo Insights: Mastering Bioluminescent Imaging for Dynamic Biological Studies

Join this webinar to learn about the three basic elements of any in vivo bioluminescent imaging experiment: the bioluminescent reporter protein, the reporter substrate, and the instrument you will use to measure the bioluminescent signal.

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Blog

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

Publication

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

Bioluminescent Imaging in Live Cells

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.

Live Cell Imaging System

To identify relevant biological processes, capturing images of live cells is a must. The GloMax® Galaxy Bioluminescence Imager combines the strength of bioluminescent reporters with the information acquired through live imaging. It is designed to enable imaging of NanoLuc® Luciferase technologies such as NanoBRET, NanoBiT, HiBiT and Lumit.

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Detecting protein:small molecule interactions with NanoBRET® NanoGlo® Detection Systems. NanoBRET® Technology utilizes a bioluminescent donor and fluorescent acceptor to visualize complex protein: protein interactions, such as target engagement. HCT116 cells expressing a PRMT5–NanoLuc® fusion were supplemented with a fluorescent small molecule tracer (top panel). Before tracer addition, luminescent signal indicates energy is present on the donor protein (left; 3-minute exposures for 15 minutes). Binding of fluorescent tracer results in energy transfer and fluorescent signal (right; 3-minute exposures for 60 minutes). Videos were captured on the GloMax® Galaxy Bioluminescence Imager.

Live Cell Substrates

NanoLuc^® Luciferase 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. In addition, its small size makes it less likely to perturb the normal biology or functionality of the fusion partner. Our Nano-Glo^® Extended Live Cell Substrates have increased signal stability, allowing extended kinetic analysis lasting several hours to days.

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Targeted protein degradation over time. HEK293 cells expressing endogenous HiBiT-tagged GSPT1 and stably expressing LgBiT were treated with CC-885 degrader or DMSO control treatment. Assayed with Nano-Glo® Vivazine™ Live Cell Substrate and imaged over 5 hours using GloMax® Galaxy Bioluminescence Imager.

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.

Additional Resources

Engineering NanoLuc^® from a Deep-Sea Shrimp Luciferase

Read this paper to learn how NanoLuc^® Luciferase was developed by merging optimization of protein structure and a novel substrate.

A Small Luciferase Brightening Up the Field of Bioluminescence

This review article details the advantages of NanoLuc^® technology for the scientific community.

nanoluctechnologysupage-imaging-dual-luciferase

Visualizing Ligand Binding to a GPCR Using NanoBRET

Read this article to learn how researchers directly quantified ligand binding to NanoLuc^®-tagged β₂-adrenoceptors in a mouse model of breast cancer.

Learn More About Imaging

Imaging

Learn about various imaging techniques, including live cell imaging, in vivo imaging, bioluminescence imaging and fluorescence microscopy.

In Vivo Imaging

Learn about in vivo imaging techniques including bioluminescence and fluorescence imaging, advantages, challenges, and applications in research.

Live Cell Imaging

Learn about live cell imaging techniques, advantages and applications.