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Sorting cells with real-time imaging: a flow cytometry game changer

Researchers can find and understand new cell populations by combining traditional flow cytometry parameters with BD CellViewTM Image Technology. Credit: BD Biosciences

Flow cytometry is a crucial technique for many scientists because it allows them to rapidly detect, sort and analyse cells with particular characteristics. But traditional flow cytometry can be a black box, where researchers are unable to directly observe their samples. Image-based flow technology adds an extra dimension of information and can open new research methods. Gert Van Isterdael has been intensively testing the new BD CellView™ Image Technology with scientists at the Flanders Institute for Biotechnology (VIB).

What is the VIB Flow Core facility and who uses it?

VIB has set up several core facilities that are shared by the institute's different research centres, and Flow Core is one of them. Many PhD students, postdocs and research technicians at VIB use the Flow Core to sort or analyse their cells, and we also welcome researchers from other institutes, universities, and industry to access our state-of-the-art technologies. We have few staff members compared to the number of machines we have, so we put a lot of effort into training users to become dedicated flow cytometry experts. The facility is open 24 hours a day, 7 days a week — and with machines becoming more user-friendly, we can train users to sort cells themselves rather than us doing it for them.

How do you contribute to the cutting-edge research?

We have intense collaborations with the VIB research teams that develop new technologies: they push us to stay on top of our game. We also closely interact with other core facilities. For example, we are part of a single-cell pipeline: users first talk to the team in the Single Cell Core to set up their experiments, then they come to us at the Flow Core to have their cells sorted. Next, the cells are loaded onto single-cell devices, such as the BD Rhapsody™ Single-Cell Analysis System, for profiling and analysis. Finally, they go to the Sequencing Core for proteomic and genomic insights. This whole pipeline is a cross-core workflow, and for users that’s a tremendous advantage.

What are the limitations of traditional flow cytometry?

Flow cytometry was always a bit blind because you can't see exactly what is going on. Is your dye on the surface, or is it intracellular? Do you have nuclear expression? Have your particles internalized in the cell, or are they sticking to the outside? It’s not possible to answer questions like this with traditional flow cytometry.

How did you find out about this new type of flow cytometry?

I network with peers, talk to colleagues, interact with VIB Tech Watch team – who scout for emerging, impactful tech – and read the latest breakthrough papers. Our users also bring ideas and inform us of novel research lines they've seen at scientific conferences in their field. They have told me about new dyes and new protocols, for example. It's a crucial interplay, so you will often find me in the lab, talking with the researchers to spark ideas. I also attend international meetings to keep an eye on what's going on in the field and with the vendors. It was at a CYTO congress where I first saw a demonstration of CellView, which uses fast imaging technology for cell sorting. We were intrigued and thought about the impact this could have at our own facility. In late 2019, we joined as one of the test sites for the prototype sorter and we started testing in April 2020.

How has BD CellView allowed you to take on challenging projects?

The technology combines high-speed imaging with flow cytometry, which makes it possible to get image data in real time. It is incorporated into flow cytometers, and uses the flow cytometers’ own components to image cells. We’d never had access to such a device before. When we got CellView, we were surprised by the speed. It was imaging more than 10,000 flow events per second. It's quite spectacular to see. We discussed how we could use this technology with our researchers and the BD applications team. Not all our ideas worked, but for many it really made an impact.

Checking cellular health becomes very straightforward, for example. The moment the cell is not happy anymore, you can see something happening on the surface of the membrane.

Do you have an example of when BD CellView imaging improved the sorting outcome?

Researchers from the VIB-UGent Center for Plant Systems Biology use our core facility to sort plant protoplasts. These cells are created by enzymatically digesting the plant cell walls, but the samples often still contain pieces of cell wall and cell debris. This creates a lot of scattering when you look at them on a traditional flow cytometer. With CellView's immediate imaging technology, we can take advantage of the fact that protoplasts are spherical, unlike cell debris, and make sorting decisions based on observable shape on the fly. We're now using the same principles to improve sort outcomes for other types of cells and generate higher quality datasets. It's a game changer.

How will this technology change the future of Flow Core?

If we combine CellView imaging technology with additional flow-cytometry parameters, we will be able to sort based on qualities such as nuclear translocation, or interacting cells. This wasn’t possible before. We've always had a very traditional way of gating — identifying and refining cell populations — but now we can find new cell populations and define new gating strategies. This will change everything and I'm 100% sure that it will push scientists to initiate new kinds of cell research.

What motivates you to go into work every day?

I'm in love with the technology. When I started working in flow cytometry about 15 years ago, I was immediately hooked. Secondly, I want to make sure that we keep everything quality controlled for our users, and that we offer good services so they can do their amazing research. I like seeing what young, brilliant minds achieve with our equipment. And finally, being able to train the current and the next generation of scientists is what drives me — it gives me energy.

What excites you about the future of flow cytometry?

I think we are about to enter a new era, where different types of measurements can be carried out by one device. On top of that, machine learning, AI and fast computing will increase automation. The combination of all of this will change the way we do flow cytometry. Whatever the future will bring us, we will need to keep training our users to keep them up to date. But with more colours and better imaging, the future of flow cytometry looks bright.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Gert Van Isterdael’s team uses BD CellView™ Image Technology for refining flow cytometry methods. For more information about how BD Biosciences can help your research, visit their webpage.

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