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Learning the FACS

Modern scientific research is being revolutionised by incredibly powerful new technologies: machines which can read your entire genetic code; microscopes which can see individual molecules inside living cells; and computers which can re-create the big bang. In this post, Lucas Greder in Marella de Bruijn’s lab describes his experiences with another such technology: fluorescence activated cell sorting (or FACS), and how learning to master this technique is critical to his ongoing PhD research.

As a graduate student, the pace at which new technology is developed and the sheer amount of it that is constantly being produced is intimidating to say the least. The vital role that technology plays in modern research requires a fundamental understanding of not only the science but also these cutting-edge methods and machines.

Luckily for researchers at the WIMM, the Institute provides opportunities for researchers to bolster conceptual understanding of the science while gaining hands on training with new technology.

One technology at the core of many projects in the WIMM is Flow Cytometry, or Fluorescence Activated Cell Sorting (FACS). As described by flow cytometrist Kevin Clark in his previous WIMM blog post, flow cytometry is used to isolate cells based on the composition of the structures that are expressed on the surface of the cell.

FACS2Our lab uses flow cytometry to interrogate early events in blood cell formation. Flow cytometry is key to our research because it allows isolation of single cells, which we then analyse individually or together with a pool of other “like” cells.

From these single cells we perform various analyses with the goal of determining how these cells are changing individually. The information gathered from these studies is then used to help figure out how to mimic the formation of blood stem cells in a culture dish in the lab. Many other groups in the WIMM also use flow cytometry as a fundamental part of their work, such as recent research by Sten Eirik and Claus Nerlov described in this blog post.

When you consider that the WIMM’s flow cytometry core serves over 300 researchers, it can be sometimes difficult to schedule time on these highly specialized machines when you need them the most. In addition, the nature of our experiments often requires late night use of the flow cytometers when operators are unavailable. These two factors pose potential impediments to the progress of our research.

A solution offered by Becton Dickinson (BD), the manufacturer of the majority of flow cytometers at the WIMM, provides researchers with a weeklong training course at their European Headquarters in Belgium. In addition to lectures on FACS theory and best practices, we learned how to run a FACSAria Fusion, the newest version of the original Aria that was first introduced in 2003. Now, our lab is able to sort cells when our hearts desire, or better, whenever our blood is being formed.

Technological advancements in flow cytometry include the ability to look at up to 15 structures on the surface of the cells at one time and do so with such sensitivity that researchers can sort cells based on fluorescence generated from simple protein-protein interactions. This presents great opportunities but requires a solid understanding of how a cytometer actually works and how fluorochromes interact with each other.

BD’s training course made sure that we had the requisite knowledge needed to fully maximise use of the flow cytometry in our research. I am particularly thankful for the expertise shared by BD and for the WIMM’s financial support to attend the course; and of course for the help provided by the original WIMM FACS man himself, Kevin Clark.

Post edited by Bryony Graham, Kevin Clark and Marella de Bruijn.


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