Data to Decision: Live Cell Imaging and Analysis of 2D and 3D Cell Models
In the pursuit of biologically relevant insights, the Incucyte® Live-Cell Analysis Systems empower scientists with simple workflows, powerful software, and unmatched throughput.
Capture high-resolution fluorescence and bright-field images of 2D and 3D cell cultures in real time over hours, days, or weeks, directly from the incubator.
The new Incucyte® CX3 Live-Cell Analysis System, featuring advanced confocal fluorescence imaging, expands assay capabilities for comprehensive studies of organoids and spheroids, accelerating your journey from data to decision. What will you discover first?
Empower Your Discovery of Complex, Multi-Cellular 3D Models with Kinetic Imaging at Scale
The Incucyte® CX3 Live-Cell Analysis System delivers cutting-edge capabilities for 3D cell culture applications. Building on the core benefits of the Incucyte® portfolio for non-perturbing, continuous analysis, the CX3 integrates confocal fluorescence imaging to enable comprehensive monitoring of multicellular 3D cell models throughout discovery and development.
Key Benefits:
- Confocal Fluorescence Imaging: Minimizes photobleaching and phototoxicity effects to capture clear images of live 3D cultures
- Industry-Leading Throughput: Enables real-time analysis of up to six microplates in parallel, maximizing data generation
- Streamlined Workflows: Intuitive workflows with integrated data analysis simplify the journey from complex cell biology to clear insights
Non-Invasive Monitoring
Reduce photobleaching and phototoxicity with patent-pending compact spinning disk technology all while operating within the controlled environment of an incubator
Unmatched Throughput
Process multiple 3D models in parallel, up to six microwell plates, for maximum productivity
Enabling Software
Unlock 3D biology with ease using intuitive software and automated processes to acquire and analyze complex biology
Kinetic, Real-Time Data
Analyze the 3D maturation and 2D growth over time, capturing key biological processes like proliferation and morphological changes.
Live Cell Imaging and Analysis Solutions
The Fast Lane to Live-Cell Insights
Derive meaningful data with sensitive, real-time live cell measurements made inside the stable condition of the incubator.
Spend more time investigating and less time troubleshooting with lab-tested protocols and purpose-built, intuitive software
Generate data rich information from every sample with fluorescent reagent combinations – compatible with a variety of cell culture models and applications
Maintain your cell health with non-perturbing image-based analysis and proprietary reagent formulations.
Make new discoveries with a wide range of applications — from complex immune-tumor cell interactions, synaptic activity in neuronal co-cultures, metabolism in cancer cells, and much more.
Learn About Live-Cell Imaging & Analysis
Live-Cell Imaging and Analysis: Revolutionizing Cell Biology
Introduction to Live-Cell Imaging
Live-cell imaging has become essential for studying the dynamic biology of living cells. Unlike single-timepoint assays, it enables real-time observation of health, behavior, morphology, and function as cells respond to their environment. The integration of 3D cell culture further enhances this approach by mimicking the structural complexity of living tissues, offering deeper insights into development, disease, and therapeutic response. As research demands grow, high-throughput, automated live-cell analysis helps streamline workflows, reduce errors, and improve data reliability.
Explore the resources below to learn more about how live-cell analysis outperforms other methods.
- Explore the complexities in continuous live-cell research.
- Discover methods and technologies for downstream analysis.
- Find strategies to minimize variability and common errors.
- Access advice to enhance your live-cell imaging outcomes.
Live-cell imaging of T cells engineered to target and kill GFP labelled tumor cells. Video acquired in Incucyte® Live-Cell Analysis System.
The realization of these benefits is not easily achieved, due to the complexities of live-cell microscopy methods involving a multidisciplinary approach that are often poorly understood and/or controlled, resulting in data that is highly variable. More challenges arise when considering the complexity that automation and incubation, which are critical for long-term live-cell imaging, add to the process.
The goal of this content is to serve as a resource for anyone wanting to learn more about long-term, live-cell imaging. Topics include:
- The advantages of live-cell imaging compared to other methods of cell analysis
- The challenges in performing long-term studies of living cell biology
- The methods and technologies used in live-cell microscopy that support downstream cell analysis
- How to reduce variability and other common failure modes when automating the process of live-cell imaging
- Practical tips and tricks to enhance the likelihood of success
Faster Assay Optimization
Real-time observation allows researchers to quickly identify and rectify issues in experimental protocols and assay set-up. This rapid feedback loop accelerates the optimization process, ensuring that assays are fine-tuned for accuracy and efficiency in a shorter time frame. The ability to make immediate adjustments based on live data reduces the time and resources spent on trial-and-error, leading to more streamlined and effective experimental designs.
Better Control of Confounding Variables
Continuous monitoring of live cells helps in identifying variables that could potentially confound the results. By controlling these variables, researchers can ensure that their findings are more reliable and reproducible, leading to more robust scientific conclusions. This level of control is particularly important in complex biological systems where multiple factors can influence outcomes, making it easier to isolate the effects of specific treatments or conditions.
Minimization of Focusing Errors and Image Artifacts
Advanced imaging systems used in live-cell imaging are designed to minimize common issues such as focusing errors and image artifacts. This ensures that the data collected is of high quality, reducing the need for extensive post-processing and increasing the reliability of the results. High-quality images are crucial for accurate analysis, as they provide clear and detailed visual information that can be quantitatively assessed.
Environmental Control
Maintaining the health and viability of cells over extended periods requires precise regulation of environmental conditions. This includes controlling temperature, humidity, and gas composition to mimic the natural conditions in which cells thrive, thereby ensuring that the cells remain healthy and active throughout the course of the experiment. Advanced environmental chambers and incubators are essential for creating stable and consistent conditions that support long-term cell culture.
Automation
Long-term live-cell imaging often involves repetitive tasks that can be efficiently managed through automation. Robotic systems can handle tasks such as media changes, cell feeding, and image acquisition, ensuring consistency and providing researchers the bandwidth to focus on data analysis and interpretation. Automation not only improves efficiency but also reduces the risk of human error, leading to more reliable and reproducible results.
Imaging Artifacts
Long-term imaging can introduce various artifacts, such as phototoxicity and photobleaching, which can affect cell behavior and data quality. Careful optimization of imaging parameters and the use of advanced imaging techniques can mitigate these effects, preserving the integrity of the cells and the accuracy of the results.
Advanced Software Tools
Specialized software is vital for optimizing long-term live-cell imaging workflows. Purpose-built applications can integrate environmental monitoring, automated imaging, and data analysis, providing a seamless experience for researchers. These software solutions often include advanced features such as real-time image processing, customizable analysis, and detailed reporting tools. By leveraging purpose-built software, researchers can efficiently manage complex datasets, automate routine tasks, and gain deeper insights into cellular behavior, ultimately enhancing the overall quality and reproducibility of their experiments.
Data Management
The volume of data generated during long-term imaging experiments can be substantial, posing challenges in storage, processing, and analysis. Efficient data management systems and advanced analytical tools are essential to handle this influx of information, enabling researchers to extract meaningful insights without being overwhelmed by the data.
High Contrast Images
Fluorescence microscopy enhances the visibility of cellular components by using fluorescent dyes or proteins that emit light when excited by specific wavelengths. This results in high-contrast images that clearly delineate different structures within the cell, making it easier to study their interactions and functions. The ability to selectively label and visualize specific molecules provides valuable insights into cellular processes that would otherwise be invisible.
Quantitative Measurements
Fluorescence microscopy allows for precise quantitative measurements of various cellular processes. By tagging specific molecules with fluorescent markers, researchers can track their movement, concentration, and interactions within the cell, providing valuable data for understanding cellular mechanisms. Quantitative fluorescence imaging enables the measurement of dynamic changes in real-time, offering a deeper understanding of cellular behavior.
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Rapid Image Acquisition
The ability to quickly capture images is crucial for studying fast kinetics in cellular activities. Fluorescence microscopy systems are designed to acquire images at high speeds, enabling researchers to observe rapid changes and dynamic processes in real-time. This capability is essential for capturing transient events and understanding the temporal dynamics of cellular functions. However, over-collection of images can lead to excess data, increasing processing time and storage demands without necessarily enhancing biological insight. On the other hand, under-collection may result in missing key events or important transitions, leading to a loss of critical biological information. To ensure accurate representation of cellular dynamics, it's important to optimize acquisition speed, resolution, and time intervals based on the specific biological phenomena being studied.
Importance of Assay Optimization for Live-Cell Experiments
Optimizing assays for extended kinetic experiments is key to maintaining cell health and ensuring high-quality, reproducible imaging. Key factors include using the right media formulation (e.g., low-riboflavin media to reduce fluorescence), filtering media for purity, and ensuring proper well volumes (50-200 µL for 96-well plates, 40-80 µL for 384-well plates). Regular media exchanges every 2-3 days for long assays are essential, unless restricted by the assay. Cell density should be optimized to prevent contact inhibition, and cells should be allowed to settle post-seeding for even distribution. Surface coatings must be tailored to promote cell adhesion, and proper vessel handling, positioning, and condensation management are crucial for preventing image distortion. Additionally, care should be taken to minimize bubble formation to avoid contamination and maintain focus.
Download Technical Note on Live-Cell Analysis Tips and Tricks
Define the Measurement
Clearly defining the objectives and parameters of the study is the first step in setting up a live-cell imaging experiment. This involves determining what specific cellular processes or behaviors will be observed and measured and establishing the criteria for success. A well-defined experimental plan ensures that the study is focused and that the data collected is relevant and meaningful.
Optimize Images for Analysis
To ensure the best possible image quality, researchers must optimize various settings such as exposure time, focus, and illumination intensity. This optimization process is crucial for obtaining clear and accurate images that can be reliably analyzed. Proper image optimization reduces noise and enhances the signal, making it easier to extract quantitative information from the images.
Automation for Image Acquisition and Processing
Automated systems in live-cell analysis streamline image acquisition and processing, enabling efficient, accurate, and reproducible results. These systems use high-definition microscopy to capture detailed, time-lapse images of cellular processes while incorporating features like noise reduction and artifact correction to improve image quality. By minimizing manual intervention, automation enhances consistency and saves time, making it essential for studying dynamic biological phenomena.
Understand Tradeoffs in Image Acquisition
There is often a tradeoff between image resolution, acquisition speed, and cell health. High-resolution images provide more detail but may require longer exposure times, which can be harmful to cells. Researchers must balance these factors to obtain useful data while maintaining cell viability. Understanding these tradeoffs is essential for designing experiments that yield high-quality data without compromising cell health.
The discovery of the cell and the development of microscopes have profoundly impacted human health and quality of life. Live-cell imaging continues this legacy by providing deeper insights into cellular processes, leading to advancements in medical research and biotechnology. This technique has the potential to revolutionize our understanding of diseases, drug development, and cellular functions, ultimately contributing to improved healthcare outcomes. By enabling the study of living cells in their natural environment, live-cell imaging offers a more accurate and comprehensive view of biological processes.
We are at a very exciting time in the world of science thanks to the advancement of AI software and the utilization of patient-derived samples. The future of live-cell analysis holds immense potential:
AI and Predictive Software
The integration of AI and machine learning in live-cell imaging will enable quicker and deeper data mining. Predictive software could potentially determine biological outcomes before they are even visible, revolutionizing the way we understand and treat diseases. AI algorithms can analyze vast amounts of imaging data, identifying patterns and correlations that may not be apparent to human observers. This capability could lead to the development of predictive models that forecast cellular responses to treatments, enabling more targeted and effective therapies.
3D Sample Analysis at Scale
The continued development of live-cell technologies will allow for the interrogation of 3D samples at scale. This will provide more comprehensive insights into cellular behavior and interactions within complex tissue structures. 3D cell cultures and organoids more accurately mimic the architecture and function of real tissues, offering a more relevant model for studying disease and drug responses. High-throughput 3D imaging systems will enable large-scale screening of these models, accelerating the discovery of new treatments.
Accessibility of Technology
Making advanced live-cell imaging technology accessible to all researchers is of critical importance. By democratizing access to these tools, we can ensure that a wider range of scientists can contribute to and benefit from the advancements in cell biology. Efforts to reduce the cost and complexity of live-cell imaging systems, along with the development of user-friendly software, will help to make these technologies more widely available. Increased accessibility will foster collaboration and innovation, driving progress in the field.
Personalized Medicine
The ultimate goal is to use live-cell analysis to determine what treatment will effectively cure a patient’s disease. By leveraging innovative imaging and cutting-edge analysis techniques, we can move towards personalized medicine, where treatments are tailored to the individual needs of each patient. Live-cell imaging can provide real-time insights into how a patient's cells respond to different treatments, enabling the selection of the most effective therapy. This approach has the potential to improve treatment outcomes and reduce the trial-and-error process in clinical decision-making.
Live-cell imaging is a powerful tool in cell biology, offering unique advantages and presenting specific challenges. By leveraging fluorescence microscopy and ensuring proper environmental control and automation, researchers can gain valuable insights into cellular dynamics, ultimately advancing our understanding of life at the cellular level. The future of live-cell analysis promises even greater advancements, driven by AI, 3D sample analysis, and increased accessibility, paving the way for personalized medicine and improved healthcare outcomes. As we continue to push the boundaries of what is possible with live-cell imaging, we move closer to a future where we can predict and treat diseases with unprecedented precision and efficacy.
Live-Cell Imaging and Analysis Handbook
The Live-Cell Imaging and Analysis Handbook is a comprehensive reference guide for the live-cell analysis technologies, focusing on the complete suite of Incucyte® Live-Cell Instruments, assays, reagents, and integrated software analysis tools. This publication is updated annually to reflect the latest Incucyte® offerings.
Live-Cell Imaging and Analysis Applications
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Cell Monitoring & Workflows
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Live-Cell Imaging and Analysis FAQs
Sartorius Live-Cell Imaging and Analysis FAQs
Get answers to the most common questions about about Sartorius image and analysis solutions including critical information regarding instruments, applications, software, service, and more.
Live-cell imaging and analysis is the combination of microscopy techniques and powerful analysis software that allows researchers to study spatio-temporal events within cells in real time. The Incucyte® Live-Cell Analysis System automatically monitors cells for days, weeks or even months as they sit stationary in the stable tissue culture incubator environment.
Fixed cells have been preserved by a fixation step that “locks” the cell in place. They provide a static snapshot into cellular function. While cellular composition is compromised, larger structures such as proteins, organelles and DNA are conserved. The fixation process kills the cells, which makes the outside cell membrane more permeable than living cells. As such, it becomes easier to target internal cellular structures with dyes and antibodies. Imaging fixed cells allows high-resolution imaging of sub-cellular structures.
However, fixed (dead) cells cannot provide dynamic insights into biological function and do not represent living systems. Live-cell imaging occurs while cells are alive. If imaged over time, real-time dynamic data can be collected and events that could be missed with endpoint, fixed cell assays are captured.
The importance in live-cell imaging and analysis lies in the ability to resolve both spatial (through higher resolution) and temporal (through time-lapse imaging) information in cells. Live-cell imaging and analysis provides dynamic insights into the health, morphology, movement and function of cell models. With live-cell imaging and analysis, we can monitor changes in real time, which is not possible with typical end-point assays that provide only a snapshot into cellular function. This is important in applications such as immuno-oncology where it becomes possible to see the real-time efficacy of cancer drugs targeting tumor cells, or in a cell migration assay where we can see how migration of tumor cells may be restricted in the presence of cancer drugs.
Live-cell imaging and analysis also provides the opportunity to make data driven decisions while the experiment is in progress. A researcher studying the biology of vascular or neuronal networks, for example, may wish to first establish a stable network before assessing the effects of compound treatments or genetic manipulations (e.g. siRNAs). With continuous live-cell imaging and analysis, it is straightforward to temporally track network parameters and use the real-time data to judge when best to initiate treatment regimes. The timing of adjunct studies such as analysis of metabolites or secreted proteins in supernatants can also be guided.
Live-cell imaging and analysis should be used when studying any area of cell therapeutics. Traditional end-point assays only provide single measurement of cellular events. Cells are dynamic in nature, so it is important to have the ability to image in real time to gain access to deeper biological insights. Live-cell imaging and analysis is typically used for therapeutic areas such as immunology, oncology, immuno-oncology and neuroscience to study cell health and proliferation, cell movement and morphology and cell function.
Images are typically visualized using microscopy techniques. Optical microscopes are a standard tool used in most labs and can visualize structures up to ~200nm range. With advancements in modern high-resolution imaging techniques and the synthesis of fluorescent probes, it is now possible to view labeled sub-cellular structures at the 10-50 nm scale. With increasing technical advancements comes increased complexity and costs; high-resolution microscopes often require extensive training and access is often limited.
High-content screening and analysis systems are benchtop systems that combine fluorescent microscopy with analysis software to visualize and analyze real-time cellular data. These systems typically do not sit within a tissue culture incubator and so are limited in terms of environmental stability and long-term studies.
Multi-mode readers are microplate readers that can detect two or more of the following: luminescence, fluorescence, time-resolved fluorescence and absorbance. An imaging system within the multi-mode reader can also image wells during the assay. These systems again lack proper environmental controls which will impact long-term studies.
Live-cell imaging and analysis systems are designed to capture cellular events as they occur in real time. Researchers can analyze a series of data-points over time, rather than a single time-point that does not provide the full picture of what their cells are doing.
Live-cell imaging and analysis systems housed within an incubator, such as the Incucyte®, can perform real-time continuous analysis over days to months, while keeping cells in a stable and optimal environment. Many technologies such as multi-mode readers and high-content imagers do not have the ability to maintain environmental control, meaning cells are not kept at physiologically relevant conditions.
Depending on application needs, the Incucyte® is also capable of running up to six microplates in parallel, allowing more throughput than typical microscopy and single plate readers. In addition to this, the Incucyte® is designed to be user-friendly. Reagents have been optimized for each application allowing easy assay preparation and intuitive software that guides the user through each step.