Live Cell Imaging
Learn about live cell imaging techniques, advantages and applications.
What is Live Cell Imaging?
Live cell imaging is a technique used in life science research to observe living cells in real-time over a period of time. This allows researchers to observe dynamic cellular processes, such as cell division, migration, and differentiation. This is in contrast with conventional fixed-cell imaging, which only captures a snapshot in time. Live cell imaging is especially useful for examining progress in tissues that change over time, such as cancerous tissues.
Advantages of Live Cell Imaging
- Real-time observation: Study events as they occur in cells over time, without having to take ‘snapshots’ of time.
- Non-invasive monitoring: Typical live cell imaging allows observation without significant harm to cells.
- Easily multiplex: The use of different fluorescent and bioluminescent markers enable the observation of several targets simultaneously within the same sample.
Disadvantages of Live Cell Imaging
- Risk of photobleaching and phototoxicity: Long exposure times may result in changes in signal.
- Markers can interfere with normal cellular function: The presence of foreign molecules or larger tags may alter cellular processes over long periods of time.
- Depth penetration: With fluorescent microscopy, signals generated deep within tissues may be difficult to discern.
Fluorescent vs Bioluminescent Live Cell Imaging
Fluorescent and bioluminescent reporters are essential tools in live cell imaging, as they generate the light signal that enables imaging in real time. Fluorescent reporters, such as fluorescent proteins and dyes, allow for the precise localization and monitoring of specific molecules within cells. Bioluminescent reporters, which generate light through enzymatic reactions within the cell, are particularly useful for imaging over extended periods with minimal phototoxicity and background interference. Learn more about their advantages and disadvantages below:
- High sensitivity due to strong signal intensity.
- Wide range of available fluorescent proteins and dyes.
- Can be used for multiplexing.
- Requires an external light source, which can cause photobleaching and phototoxicity.
- Autofluorescence from cells can lead to background noise.
- Photobleaching can reduce signal over time, limiting long-term imaging.
- Signals generated deep within tissues may be difficult to discern due to light scattering.
- No need for external light source, reducing background noise.
- Suitable for long-term imaging due to low phototoxicity.
- Can be used in deep tissue imaging due to lower light scattering.
- Generally lower signal intensity compared to fluorescence.
- Limited options for color variants compared to fluorescence.
- More complex and expensive to develop and optimize.
What is a Live Cell Imaging System?
Live cell imaging systems are specialized instruments that include a microscope, environmental control and image acquisition software. This combination allows cells to maintain viability while being imaged over a long period of time.
The GloMax® Galaxy Bioluminescence Imager combines the strength of bioluminescent and fluorescent reporters with live imaging capabilities. It is designed to enable imaging of our NanoLuc® Luciferase technologies (e.g., NanoBRET, NanoBiT, HiBiT and Lumit) to visualize various cellular processes:
- Protein:protein interactions
- Protein localization and translocation
- Protein degradation and stability
- Ligand:protein interactions (target engagement)
- Targeted cell killing

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. 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.
What Happens in Real Time, and Why It Matters for Drug Discovery
Webinar: Real Time Live Cell Imaging and Analysis for Drug Discovery.
See how live-cell kinetic reagents, multiplex data, and imaging simplify drug toxicity testing and reveal apoptosis dynamics.
What is Bioluminescent Live Cell Imaging?
Bioluminescent live cell imaging allows direct visualization of protein dynamics in living cells without the need for repeated sample excitation.
How does bioluminescence work?
NanoLuc® Luciferase reporters are well-suited for use in bioluminescent imaging 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. Its small size also 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.

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.

Dr. Li-Fang Chu's lab observed the exact timing of oscillating genes in early human development for the first time using bioluminescent live cell imaging. Read more in this blog
What is Fluorescent Live Cell Imaging?
Fluorescent Ligands for Live Cell Imaging
We offer a large variety of fluorescent HaloTag® Ligands that enable live cell imaging. Explore our full suite of fluorescent ligands.
Fluorescent Live Cell Imaging Protocol
Interested in fluorescent live cell imaging, but not sure where to start? We provide comprehensive protocols for both rapid and no-wash live-cell imaging using our fluorescent HaloTag® Ligands. View the protocol to learn how to optimize a signal that is both robust and specific.

What You See at the Single-Molecule Level — And Why It Changes Live-Cell Imaging
Webinar: Using HaloTag® Technology and Janelia Fluor® Dyes for Live Cell Single-Molecule Imaging
Discover how this technology uncovers protein dynamics in native environments, enabling better decisions, sharper insights, and reproducible results.
How Live Cell Imaging Is Used Across Research Applications
Cancer Research
Live cell imaging allows for the tracking of tumor cell behavior, including growth, invasion, and response to treatments. Cancer-specific markers and pathways can be monitored in real time using live cells.
Developmental Biology
Developmental processes like cellular proliferation, replication, and tissue formation are well-suited for study through live cell imaging. Understanding these processes requires visualizing the spatial and temporal patterns of gene expression and protein localization.
Immunology
Live cell imaging allows for the visualization of immune processes, including immune cell interactions, signaling pathways, and the immune response to pathogens. It can closely monitor inflammatory responses, such as gene activation in response to infection, and processes like phagocytosis.
Neuroscience
Live cell imaging enables the observation of neuronal activity, such as synaptic function and calcium signaling, within live neurons and larger neuronal networks. This technique allows for the study of neuron-specific genes and pathways that are implicated in neurodegenerative conditions over extended periods.
Microbiology
Microbial cells can be labeled with fluorescent or bioluminescent markers, enabling the study of host-pathogen interactions, biofilm formation, and microbial behavior. This approach is particularly useful for observing bacterial and viral infections, which is crucial for developing effective antimicrobial treatments.
Additional Resources
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.
Bioluminescence Imaging
Learn about bioluminescence imaging in whole animals and in live cells.