Frequently Asked Questions

The system is ideal for:

• Validating NanoLuc®-based assays (e.g., NanoBiT, NanoBRET, Nano-Glo)
• Visualizing protein localization and degradation
• Tracking live-cell kinetics over time
• Confirming assay results from a plate reader using spatial imaging.

Want to explore a full array of applications? View the GloMax® Galaxy Application Hub.

The right technology depends on the biology you want to observe:

• Use NanoBiT® technology for protein-protein interaction studies.
• Use HiBiT to monitor endogenous protein levels and localization.
• Use NanoBRET® technology to track molecular engagement or trafficking events.

These are compatible with live-cell formats and produce bioluminescent signals well-suited for imaging with the GloMax® Galaxy Bioluminescence Imager. Select an assay that gives a durable signal and minimal perturbation to the system you're studying.

• For end-point experiments: Prioritize signal intensity with Nano-Glo® Live Cell Assay System for maximum brightness when kinetic imaging is not required.
• For kinetic real-time imaging: Prioritize stability with the use of Nano-Glo® Vivazine® or Endurazine™ for tracking protein dynamics over hours.

Yes. The GloMax® Galaxy Bioluminescence Imager supports fluorescence, filtered luminescence, and brightfield imaging using modular excitation/emission components. Additional filters and modules are available to match common fluorophores. 

Bioluminescence offers lower background and doesn’t require excitation light, reducing autofluorescence and photobleaching. This is particularly helpful for long-term, live-cell imaging or applications where signal sensitivity is critical. Due to fewer photons being released, exposure times are longer with bioluminescence than with fluorescence. 

See also: Bioluminescence vs. Fluorescence

Start by defining your experimental goal—whether you're tracking protein degradation, localization, or interaction. Make sure you’re using phenol red–free media to reduce background and that your plate or dish format is compatible with the imaging stage. If you'll be doing long-term imaging, plan for low-phototoxicity conditions and consider environmental control options like a stage-top incubator. Seeding density and signal duration (e.g., long-lived substrates) should also be optimized for your time window.

Supported vessels include slides, microchamber slides, 35mm dishes, and 6- to 96-well plates. Preconfigured focus offsets are available for commonly used formats like ibidi slides and Greiner microplates.

For luminescence imaging, an exposure of 1-5 minutes should result in appropriate image capture. Up to 60 minutes maximum exposure time is possible; however, some resolution will be lost. Start with a 30 second exposure and increase exposure by 30 second intervals until sufficient signal is acquired.

• Use media without phenol red to reduce background.
• Maintain proper cell seeding density (e.g., 1 × 10? cells/mL).
• Ensure proper cell adherence to the plate/vessel bottom to improve image focus.
• Choose long-lived substrates like Endurazine™ or Vivazine™ for extended imaging windows. Consider using binning (2x2 or 4x4) to boost sensitivity for low-abundance proteins.  Binning combines neighboring pixels to boost signal brightness when imaging faint targets.

Yes. The system saves standard .TIFF files, fully compatible with ImageJ/Fiji. Fiji is included with the system and comes pre-installed on the provided PC.

• Low signal intensity: Confirm proper substrate concentration and avoid over-dilution. Use binning to boost sensitivity if needed. Test several exposure times.
• High background: Use phenol red–free media and ensure the imaging vessel is clean and properly seated.
• Cell health issues during imaging: Avoid overexposure and excitation-based methods when possible. For long-term experiments, use long-lived substrates and minimize temperature fluctuation.
• Drifting focus: Make sure the vessel matches the selected imaging protocol and is seated flat on the stage. Ensure your cells are adhered to the plate bottom with brightfield imaging. 

Want more help with your live cell imaging experiments? Consider reading our white paper: How To Image Live-Cell Assays on GloMax® Galaxy Bioluminescence Imager

 

The GloMax® Galaxy Bioluminescence Imager is a Research Use Only Device. Not for Use in Diagnostic Procedures.