3 Factors to Consider for Automated Live-Cell Imaging

Live-cell imaging is important for many applications however limitations of conventional methods have constrained its routine use. Reliable live cell imaging requires an environment that keeps the cells functioning during the experiment while also being able to ensure the experimental method is not perturbing the cells and affecting the interpretation of the results. Here we discuss the growing popularity of automated live-cell imaging systems and highlight some key features to look for when selecting a live cell imaging system.

What is live-cell imaging?

Live-cell imaging is a microscopy-based technique used to examine living cells in real-time. It offers deeper insights into dynamic cellular processes such as migration, confluency and signaling and can reveal findings that might otherwise have been overlooked. Both brightfield and fluorescence-based live-cell imaging modalities support a range of different analysis needs.

How is live-cell imaging used?

Applications of live-cell imaging span basic research through to biopharmaceutical manufacturing. In a research setting, live-cell imaging can be used during cell culture to help define the best time for harvest, determine the senescence status of cells, assess drug treatments for cytotoxicity or detect and monitor phagocytosis. For example, Phagocytosis, is a process by which certain live cells, called phagocytes internalise foreign matter. This defensive reaction against infection is key in the study of immunology and plays an important role in immune responses, tissue homeostasis, and continuous clearance of apoptotic cells. Generally, phagocytotic activity is assayed using flow cytometry. However, this process only provides quantitative data and does not provide the means to monitor phagocytosis in real time. Performing fluorescence-based assays using the CELENA® X High Content Imaging System with pH-sensitive fluorescent particles, like pHrodo™ Green, can be an effective and efficient system for quantifying and monitoring apoptosis activity. For biopharmaceutical manufacturing, live-cell imaging has broad utility for process development and control throughout the production of biologic drugs and vaccines.

Limitations of conventional live-cell imaging methods?

Historically, live-cell imaging has involved manually monitoring cells by culturing them in a CO2 incubator. The culture vessel is removed several times to take images of cells over time using a digital microscope. This approach is labor-intensive and highly prone to human error, largely because it offers no means of finding the same position in the culture vessel. Fluctuating environmental conditions can also cause cellular stresses, which can compromise results. While benchtop imaging systems improve on this method, they are bulky and cumbersome, and often struggle to maintain a stable environment.

3 Factors to consider when choosing an automated live-cell imaging system

Automated live-cell imaging systems like the CELENA® X offer a flexible design that is smaller, faster and easier to use to meet both the demands of the drug discovery industry and the basic research needs of the smaller laboratory.

Multiple imaging modes that are affordable

Live cell imaging systems that offer both brightfield and fluorescence options for either time-lapse or real-time monitoring offer maximum flexibility. Celena X integrates an automated fluorescence microscope with quantitative image analysis software to process large datasets at an affordable cost. Its interface allows a user to run multi-well or multi-spectral experiments with capacity for multi-point imaging with only a few clicks. The microscope provides for a multitude of fluorescence cell imaging possibilities by supporting all objective magnification from 1.25x to 100x, both brightfield and phase contrast illumination, and with LED filter cubes.

Stable scanning performance and compatibility

The system is compatible with a wide range of cell culture vessels such as multi-well plates, dishes, flasks and slides to cover a wide variety of assay types. Depending upon the application, either image-based or laser-based autofocus methods can be used. The CELENA® X can be used for image confluency in McCoy cells seeded in 96-well plates over 48 hours with brightfield image-based autofocusing, demonstrating how the system can be modified and applied as a high throughput method for various cell-based assays. With laser-based autofocusing and multiple filter cubes, this method utilised the CELENA® X to image the dose-dependent effects of anti-cancer drugs throughout the cell cycle in HeLa cells seeded in 96-well plates, demonstrating how the system can be used in a multivariate drug screening process.

User friendly interface and 3D modeling

An analysis of high content images with large datasets can cause problems with most types of analytical software. Each new assay requires the creation of new modules, which can be challenging for lab staff and often involves IT staff. The CELENA® X provides an easy-to-use modular-design analysis software based on a powerful CellProfiler engine. Tens of thousands of images can be analysed automatically to obtain quantitative information with a complex software setting or optimisation.

Three-dimensional (3D) cell models can provide a more accurate representation of real cell environments than 2D cell culture systems but requires a different strategy of imaging and analysis compared to 2D cell culture. Organoids, for example, are organ-specific 3D cell models derived from human stem cells, designed to mimic the functionality and structure of human organs while 3D spheroids can represent a gradient of nutrients and oxygen between cells located in both outer and inner layers which is more relevant to physiological environments. These 3D models are notably useful for studying various types of cancers.

The challenge when imaging organoid and spheroid assays comes from organoids having multiple focal planes making it difficult to acquire in-focused images for multiple organoids. For live/dead cell viability of the single organoid, a different analysis strategy is required since individual cells in an organoid do not exist as a single live/dead status. To address this issue, CELENA X employs MergeFocus software module after acquiring Z-stack images from multi-channel fluorescence.

We invite you to use the Celena X automated live cell imaging system today and compare it for yourself.

Contact us to arrange a free trial.

ATA Scientific Pty Ltd
+61 2 9541 3500
enquiries@atascientific.com.au 
www.atascientific.com.au 

Product page link: – https://www.atascientific.com.au/products/celena-x-high-content-auto-cell-imaging-system/