Essen Bioscience

IncuCyte™ live-cell imaging systems, designed by Essen BioScience is the first instrument to provide automated real-time image capture and data analysis from hours to weeks, directly inside your incubator. The IncuCyte™ systems also offer a variety of software modules as well as in-vitro assays, reagents, cell lines, consumables and customized services that target functional measurement of living cells over time.

Gain insight into active biological processes, in real-time

Few would argue the added insight that the kinetic measure provides - the opportunity to observe the full signal time-course and then select optimal time points for post hoc analyses.  The chance to follow the sequence of biological events and to study dynamic processes. For image-based experiments in living cells, the chance to rewind and replay the experiment to observe what really happened to your cells whilst.

Then why do investigators often blindly select single time- and end- point measures at all?  The  answer is more a practical than a biological one - temporal measures take more effort to make than end points and scientists with home-lives and sleep requirements favor doing so over making measurements in the laboratory at anti-social hours.

Quantitative time-lapse live-cell imaging is one method with the promise to solve this. However, the majority of solutions for time-lapse are engineered for high spatio-temporal resolution studies in small numbers of cells in the milliseconds to hours timeframe. This is at the expense of throughput and ease of use. Generally, a high degree of imaging expertise is required and extracting measurements and decision making data from large image stacks takes time and is rate limiting.   For longer term studies (days or weeks) preservation of the cell environment (temperature, humidity, pH) is paramount and hard to achieve. The current solutions do not lend themselves to positioning live cell imaging as a frontline assay tool.

Designed to address these challenges, the IncuCyte ZOOM is a live-cell imaging and analysis system that enables quantification of cell behavior over time by automatically gathering and analyzing images. The system resides in a standard cell incubator. Equipped with high definition phase-contrast red and green fluorescence, optics from 4-20x magnification, the IncuCyte ZOOM® System scans cell plates or tissue culture flasks on a customizable duty cycle.  Imaging is non-invasive and non-perturbing to cell health. Unlike other time-lapse imaging systems, the IncuCyte can process multiple plates in parallel and does not depend on shuttling plates in and out of the incubator. A single system can house 6 micro-titer plates or T-flasks and can automatically acquire and analyze up to 2000 images per hour.

Whether it be for simple cell proliferation assays or more advanced stem cell differentiation biology, our live-cell, non-perturbing measurement approach yields real-time data and insight far beyond that achievable with conventional end-point or non-image based approaches. Our Discovery Services can also provide integrated or turnkey solutions for researchers who seek specialized, customized solutions.

IncuCyte ZOOM® System is connected to the internal IT network via a specialized controller. Acquisition schedules, image analysis and data viewing are conducted by any connected remote PC based device running the ZOOM® Software. This architecture sets the system up as a shared resource for researchers within and across laboratories and makes it ideally suited for collaborative work. Importantly, images can be gathered with ease, repeatedly, and without any requirement to prepare samples. Data is analyzed on the fly, image by image, providing a real time insight into cell behavior. Within the user interface it is simple to view temporal data from each plate and flask and to create summary views across treatment groups for the parameters of interest (confluence, fluorescent object count etc.). All cell images are archived into a date-organized, fully searchable databases meaning any image for any time-point is just a click away. Additional assay modules with tunable algorithms for assays such as migration, vascular tube formation and neurite-outgrowth increase this power. Critically, while a working knowledge of basic principles is required, assays can be run and analyzed with minimal training on the finer points of cell imaging or image analysis. 

The IncuCyte™ System revolutionizes the way researchers think about cell imaging and image-based assays. The IncuCyte can be deployed not only as a plate reader for assays but also in a mode to scan cells in the pre-experiment cell preparation phase. Unexpected changes in cell morphology and proliferation rates can be registered and considered prior to starting an experiment. Once cells are dispensed into microtiter-assay plates the IncuCyte can be used to verify that they are evenly-plated and healthy. For assays on proliferating cells, the timing for the start of treatments can be triggered from a pre-set confluence value.  Quality controlling cells and assay plates in this way helps improve assay performance and consistency by ensuring that experiments are only conducted on healthy, evenly plated cultures with the expected cell morphology. This surveillance mode with whole-well viewing is also powerful for capturing rare events - new colony formation when reprogramming stem cells or for monoclonal cell line development. In each of these examples it is not necessary to conduct a detailed analysis of every image taken.  Rather, it is more valuable to obtain a rapid, cursory and high level view and to be able, where necessary, to drill in and inspect areas (e.g. positions of emerging colonies) and times of interest.

Another mind set change concerns assay signal pre-reads. Assay plates can be analyzed for the parameter/phenotype of interest prior to the addition of treatments to provide a within well baseline measure.  This contrasts with the typical workflow for high content screening (HCS) in which plates are processed (fixed, labelled, imaged) and analyzed only at the end of the experiment.  With this end point approach, all comparisons of treatments need to be made across rather than within assay wells, which has an inherently lower statistical power.  If you do wish to determine the time course of a biological event using HCS data needs to be gathered from different wells at different time points and then concatenated to assemble the time-course – this is inefficient and unappealing, especially if the key data points are in the middle of the night or over the weekend!

The IncuCyte™ approach also provides the opportunity to make data driven decisions whilst 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 IncuCyte time-lapse imaging, it is straightforward to temporally track network parameters and use the real time data to judge when best to initiate the treatment regimes. The timing of adjunct studies such as analysis of metabolites or secreted proteins in supernatants can also be guided. Drug washout studies may be performed using the real time data to identify when an equilibrium response occurs and to trigger the timing of the washout regime. If for any reason it transpires that the experiment is not panning out as expected then treatments could be withheld to save expensive reagents and follow-on experiments can be initiated more quickly to make up time.  

Time-course data also affords greater opportunities for different and novel analyses compared to end point metrics. Area under curve, time to signal onset or threshold, and rate parameters (dx/dt) are at times highly valuable. Simply calculating the assay signal at its peak timepoint and/or at the optimal signal/background all helps in assembling robust and reproducible assays. Of course, transient effects of treatments can be detected by kinetic imaging that may otherwise be missed with end-point reads.

The combination of the real-time kinetic and image-based method is extremely helpful when developing, validating and troubleshooting phenotypic assays. Within a small number of assay plates it’s usually possible to obtain a clear understanding of the relationship over time between assay signal and treatments, cell plating densities, plate coatings and other protocol parameters. Scrutiny of the kinetic data and movies from each well help to rapidly pinpoint sources of within- and across-plate variance (e.g. uneven cell plating, poor cell health, washing artifacts) and to validate the biology of interest.  This is particularly true for more advanced cell systems such as co-cultures where far more permutations and combinations of protocol parameters exist (e.g. cell plating ratios) and the biology is more complex.  Along these lines, the IncuCyte has been used to assemble a range of more translational cellular assays. These include human model systems for angiogenesis (vascular endothelia + fibroblasts), immune-cell killing (T-lymphocytes + tumor cells), neurite dynamics (neurons + astrocytes) and the tumor microenvironment (tumor & stromal cells). Many of these assays are based on combining phase contrast (label free) image analysis with live cell fluorescent probes such as cytoplasmic or nuclear targeted GFP/RFPs for cell counting and substrate-linked DNA labelling fluorophores for enzyme detection (e.g. caspase 3/7 apoptosis reagent). The ever increasing number and diversity of non-perturbing fluorescent biosensors that are suited to long term live cell imaging is a key enabler for the range of assays that can be configured.

Whether in surveillance mode or as a kinetic plate reader the IncuCyte™ is re-defining the possibilities and workflows of cell biology. The combination of throughput, long term stability and non-invasive measurement enables researchers to monitor and measure cell behaviors at a scale and in ways that were previously not possible, or at the least highly impractical.