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:

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.

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

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.

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.

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.

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.

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.