FACS Expert Sees Blue When Testing Flow Cytometry Reagents

BD Biosciences sat down with Jochen Lamote, Head of the Vlaams Institute for Biotechnology (VIB) Flow Core in Leuven, Belgium to ask him questions about innovation in flow cytometry reagents. At the VIB Centre for Cancer Biology, they focus on immunophenotyping and cancer cell characterisation. While at the VIB Centre for Brain and Disease Research, they work to isolate specific brain cells.

Why did you agree to be a beta tester for these flow cytometry reagents?

VIB always wants to go the extra mile. We want to test new technologies, we want to go for implementation of new techniques. In this case, we are very interested in implementing the BD FACSDiscover™ S8 Cell Sorter with BD CellView™ Image Technology.

For this, we need specific dyes that are primarily excited by the blue lasers, so this is why we're very interested in testing more BD Horizon RealBlue™ Reagents.

Why is reduced cross-laser excitation so important to your research?

These dyes are very specific, and the advantage of this is that when you look at subtle changes when you need a high resolution to see the subtle changes, that we can use now these dyes instead of dyes that are having some spillover in other channels, which always leads to compensation issues, which will decrease the resolution.

For very sensitive assays, we prefer to use these dyes. So compared to other dyes, for example, the BD Horizon RealBlue™ Reagents are primarily excited by the blue laser. It was important for us to assess that, that you don't have crossover in the yellow-green laser, while, for example, phycoerythrin-cyanine7 (PE-Cy7) will be excited by both.

This was the main goal for us, to see the sensitivity and the exclusivity compared to a laser, let's say. The more distinct fluorophores are and exclusive the fluorophores are, the better for us.

How important is panel design flexibility in your research?

The other reason why we also want to have these, let's say, very distinct dyes compared to what's already available on the market is also because in academia or research goes very fast. We have a huge turnover.

Experiments change all the time. This is not like in a clinical company where a panel is designed once and optimised, and is being used then for the next 3, 4, 5, 6 months or years even. The BD Horizon RealBlue™ Reagents and the BD Horizon RealYellow™ Reagents have less issues with spillover, so it's easier to just quickly switch markers without affecting the other colors in the panel.

Have you tried using BD Horizon RealBlue™ Reagents in conjunction with other spectrally similar fluorochromes?

We recently discovered that you can use BD Horizon RealBlue™ 780 as an additional dye in your panels so you don't have to switch markers anymore in a way that we have panels which already have PE-Cy7, but BD Horizon RealBlue™ 780 is not cross excited by the yellow-green laser.

This is an advantage for us because we can just add this marker now to our panels and we can distinguish between BD Horizon RealBlue™ 780 and PE-Cy7 without any issues.

Would you recommend this new family of BD Horizon™ Reagents to your colleagues? 

We are really going to push our scientists as well to use these dyes who have less cross-laser excitation because it makes the panel design way easier especially with the blue laser and the yellow-green laser, there's a lot of cross laser excitation with the currently available dyes.

This will go away now if people really use BD Horizon RealBlue™ Reagents and BD Horizon RealYellow™ Reagents instead of the typical phycoerythrin (PE) tandem dyes that are existing.

Are you looking forward to future releases of flow cytometry reagents? 

Especially now with BD CellView™ Image Technology as well, it'll open a whole lot of possibilities again to do new unprecedented experiments. We can address now new biological questions with these technologies. Innovative technologies are super important to drive our science. For me, the BD Horizon RealBlue™ 780 really stands out.

Learn more about BD Horizon RealBlue™ Reagents and BD Horizon RealYellow™ Reagents