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By Dr Christian Oberdanner
Last night you were up until midnight tending to your live-cell experiment. This morning you woke up with great expectations, only to find that your cells are sick and the entire experiment must be repeated. Sound familiar? It happens all too often, and the consequences can be heartbreaking – deadlines missed, expensive reagents wasted, precious samples lost. Automating your live-cell assays can help prevent costly setbacks like these, but only if the solutions you implement are up to the job. With various vendors and automation options to choose from, how do you decide on the live-cell set-up that’s best for your research?
Make real-time stem cell analysis and other tricky live-cell applications as easy as child’s play, with Spark smart automation.
Learning from the experts
To address this question, we took a look at how researchers at the University of Heidelberg successfully tackled the difficult challenge of automating stem cell differentiation. The team of professor Rudiger Arnold was interested in understanding the molecular mechanisms that human mesenchymal stem cells use to switch from proliferation to differentiation. Automation played a central role in their ability to resolve and accurately quantify cellular changes during the early stages of differentiation. Based on their experiences, here are some top points to consider when choosing a live-cell automation solution that will keep both you and your cells happy:
1. Take control of the environment.
Robust environmental control is fundamental for maintaining cell health throughout extended time course experiments. Stem cells are notoriously sensitive to variability in temperature and gas concentrations, and this problem becomes even more acute when culturing in low-volume microplates. Fluctuations in the culture environment can radically impact differentiation outcomes. Even the process of transferring stem cells from an incubator to a plate reader can perturb the environment within the culture dish enough to alter cell fates.
For rigorous control in directing their cells along defined lineages in 96-well format, the Heidelberg team looked for a plate reader with fully integrated environmental control. The Spark® multimode plate reader provided them with independent regulation of CO2 and O2 levels to ensure an optimal culture environment. Active heating and cooling enabled them to maintain the strict temperature tolerances required to avoid triggering stress responses. The convenient Humidity Cassette was compatible with their preferred plate types, providing effective protection against evaporation.
2. “Hands-off” may be best.
Manual handling steps are an obvious source of experimental error and contamination in live-cell assays, especially complex protocols that run over the course of several days. Since stem cells are typically cultured without antibiotics, they are especially vulnerable to contamination. To reduce hands-on time and minimize the chances of error, professor Arnold’s team opted for a fully automated differentiation workflow. Spark’s modular design enabled them to migrate the entire process to a single platform tailored to their specific needs, such as real-time multi-parametric analysis. An integrated lid lifter and injection system allowed them to manipulate plates and add reagents when needed without risking manual contamination.
3. Get smarter with your automation.
Automation of live cell assays has come a long way – it’s no longer just about robotic plate handling and liquid transfer, but about how seamlessly tasks are integrated, and how responsive the automation is to your changing needs. Spark’s Smart Automation gave the Heidelberg team complete control over experimental design, environmental regulation and kinetic scheduling, with a straightforward user interface that let them customize their protocols easily. Conditional workflow automation (more of which in future blogs) opened up new experimental possibilities.
4. Stay flexible.
If one thing is common among researchers, it’s that our needs are always changing: today Professor Arnold’s focus is stem cell differentiation, tomorrow it could be something completely different. The Spark multimode microplate reader is equipped to handle a wide range of in vitro and live-cell assays. The modular design allows the Heidelberg team to freely configure and upgrade the reader to meet evolving needs, including the addition of cell imaging capability. Smart features cater to a multi-user environment to increase productivity. For example, a convenient pause/resume function allows peers to access the reader for other applications during the course of a kinetic run.
5. Find your sweet spot.
It’s tempting to fall for the hype that cell-based assays require expensive high-resolution imaging equipment. Without a doubt high-content imaging systems have their place in cell biology research, but the truth is that they are overkill for most cell-based assays, including those run by Arnold’s group. On the other hand, low-budget solutions like cell counters simply aren’t sophisticated enough to address complex biological questions. Arnold’s group found that Spark hit the sweet spot – offering the quality, attention to detail and flexibility they needed to handle current applications, in a fully upgradeable format that could grow with their needs.
Learn more about how Spark is helping researchers at the University of Heidelberg unravel the mysteries of stem cell proliferation during the early phases of differentiation. Download the application note here.