Cell imaging and analysis techniques have been fundamental to the advancement of biomedical sciences. From the first observation of living cells under a microscope, to time-lapse imaging of various biological processes, cell imaging techniques have evolved to provide greater spatial and temporal resolution. Despite ongoing evolution, these techniques and technologies are limited in their ability to track biological changes over longer periods of time without perturbing cells during imaging. Current methods also fail to support high-throughput applications, and are inaccessible to many researchers based on cost and complex software required for image acquisition and analysis. After 15 years of development, the IncuCyte® Live-cell Analysis System emerged as a robust, innovative solution to these challenges, with significant implications for the future of cell biology.
Maintaining Optimal Environmental Conditions
The cell culture incubator maintains stable, well-controlled conditions to mimic the cell’s natural environment. Traditional methods of analysis necessitate removing cells from the incubator, exposing them to variable temperature and atmospheric conditions, and potentially affecting experimental outcomes. Cold shock, for example, can arrest cell proliferation, while varying levels of oxygen and carbon dioxide can lead to the generation of reactive oxygen species, and alter pH, respectively.
The integration of microscopes and live-cell imaging devices into the highly controlled cell incubator environment enables continuous, scheduled image acquisition without perturbing the cells with repeated incubator opening. The IncuCyte Live-cell Analysis System combines non-invasive image acquisition with real-time image processing and data analysis to provide continuous live-cell analysis for valuable insights into cell behavior.
Better Insights Tell the Whole Story
There are numerous assays designed to assess cell health and morphology, movement, activity, and function. Traditional assays provide only limited, endpoint information pertaining to these parameters, whereas live-cell imaging and analysis provides a more complete story of the events occurring throughout an experiment.
The benefits of live-cell imaging and analysis may be illustrated as an analogy between a cell proliferation assay, for example, and a sporting event. Cell proliferation assays are often used to assess tumor cell growth, and are fundamental to the development of cancer therapeutics. Implementing endpoint analysis for quantification of proliferation does not provide sufficient information to distinguish the effects of different conditions or treatments, similar to missing the entire sporting event, only to see the final score. Alternatively, live-cell imaging and analysis is capable of distinguishing temporal differences between treatment effects, akin to watching the entire sporting event and seeing what individual events contributed to the final outcome.
The IncuCyte Live-cell Analysis System supports multiplex measurements to provide further insights into cell health—in addition to increasing productivity. Multiplexing proliferation, for example, with measurements such as apoptosis or cytotoxicity can be used to discriminate between cytotoxic and cytostatic treatment effects.
Redefining the Possibilities for Imaging Assays
The ability to study cells within the confines of the stable incubator environment has created new possibilities for imaging assays. This approach prioritizes cell health, making it possible to study various cell models, from primary cells to stem cells, and assay types ranging from 2D assays to more complex 3D assays.
Stem cells may be used to create differentiated neurons and support cells necessary for neurological disease research. Evaluation and characterization of human induced pluripotent stem cells (hiPSCs) is limited by current methods, such as immunocytochemistry, microelectrode techniques, and flow cytometry, that do not enable long-term monitoring, and potentially perturb fragile cells during sample preparation. Live-cell imaging and analysis enables non-invasive evaluation of these cells within a physiologically relevant environment, facilitating characterization.
Three-dimensional cell culture models are becoming more widely used to accelerate drug discovery, notably in cancer biology and hepatotoxicity, as they provide important biological features including hypoxic regions, and are capable of interacting with immune cells. Studying these models is challenging due to time consuming workflows, and labeling that affects cell biology, in addition to the inherent limitations of endpoint analysis. Live-cell imaging and analysis supports 3D cell culture applications, incorporating label-free techniques, non-perturbing reagents, and real-time monitoring of cell health and morphology while cells remain in the stable incubator environment.
Live-cell imaging and analysis has been applied to a wide range of cellular assays, from cell proliferation to immune-cell killing, apoptosis, and cytotoxicity, among many others. With this technology, scientists can explore beyond the confines of traditional methods by devising new experiments that were once considered impossible.
Supporting Data-Driven Decision Making
Live-cell imaging and analysis enables investigators to non-invasively observe and measure the entire cell biology workflow, from cell culture, to manipulation, to assay. Routine monitoring makes it possible to evaluate cell passage to ensure consistency, as well as proper seeding and transfection. It also supports data-driven decision making throughout the course of the experiment. Using live-cell imaging and analysis, an investigator can identify when a stable vascular network has been established prior to initiating treatment or manipulation. It also overcomes the limitations of existing technology to monitor stem cell differentiation and maturation. Using live-cell imaging and analysis, investigators can follow the development of hiPSC-derived neurons, and ensure they are functionally active prior to further downstream analysis.
The IncuCyte Live-cell Analysis System enables long-term, continuous monitoring and multiplex analysis of cells within the fully controlled incubator environment. IncuCyte technology provides researchers confidence in the accuracy and reproducibility of their data with a true representation of biological processes.