David Agorku was the lucky winner of a competition that we ran last year offering pharma and biotech companies the chance to win the use of a CX-A for 6 months.
David’s goal is to understand how T cells and fibroblast cells interact, so we provided him with a CX-A installed with our standard analysis program, EVE Analytics and a copy of our Live T Cell Assay (LTCA).
Our Scientific Communication Manager, Dr Emma Gibbin-Lameira, caught up with David recently to find out how he had used the microscope and what he thinks about our digital analysis solutions. The first question asked was how Nanolive platforms compared to the techniques he usually uses in his research.
“Normally, I co-culture T cells with different fibroblast cell lines then harvest the T cells and analyze them using flow cytometry. Although the assay works well, it’s an end-point technique which means we miss all of the dynamic interactions. In the past, I have also tried using live imaging platforms, but all were fluo- based, which meant I had to either transduce or stain my fibroblast cells. I did try this, but in the end, I did not pursue it further as the technique itself adds a margin of error into your experimental results. Having a label-free system like Nanolive’s is extremely helpful because you can take pretty much any cell line, load it on a plate, and easily analyze it without imaging artefacts”.
So, what did David think about the image analysis solutions Nanolive provide?
“The CX-A opens up a whole world of opportunities to analyze cells in co-culture. Especially the software is extremely powerful. Segmenting cells is one of the biggest challenges we face in the analysis of cells in vitro! Your software does a very good job of this, and compared to our current systems, I have to say its impressive”.
Another thing David was impressed with was the range of metrics offered by the LTCA.
“The LTCA allows you to see whether a single T cell is in contact with a target cell or not, and how much of the overall area of target cells is covered by T cells. Parameters like this offer us an amazing insight into cell interactions, that you simply cannot get them with other systems”.
As is the case with most research in the biotech and pharma domain, David’s research is confidential so we cannot reveal his results in too much detail, but he was able to provide us with some of the incredible videos he took.
Video 1 represents a typical co-culturing experiment for David. In this case, T cells (coloured red if they are not in contact with their target cells and yellow if they are) were inactivated, so despite a relatively high T cell:target cell ratio, they have little effect on their target cells (coloured in blue).
“The CX-A opens up a whole world of opportunities to analyze cells in co-culture. Especially the software is extremely powerful. Segmenting cells is one of the biggest challenges we face in the analysis of cells in vitro! Your software does a very good job of this, and compared to our current systems, I have to say its impressive”
Activated T cells do not always lead to target cell death, but they do invoke signs of stress as can be seen in Video 2. Often signs of stress show as changes in the shape of mitochondria, which become rounded. When Emma pushed David a little further to see what his most interesting finding was, he smiled and replied:
“Normally we tend to focus on T cell performance, but our experience with the CX-A showed us that there is a lot going on with the fibroblasts that we didn’t have on our radars and that we will have to dig into right now”.
Promising stuff. So, where does David see the future of Nanolive’s label-free technology?
“Seeing what is being developed in the field of personalized T cell therapy I think it would be really powerful diagnostic tool for controlling the quality of T cells. Being able to QC T cells without the need of staining either the T cells or the target cells will be extremely valuable. Nanolive imaging can also be a game-changer for developing new or better binders. The LTCA is particularly useful for this as it allows to see how T cells perform in real time and quantify which T cells bind to my target cells and how efficient my T cells are at actually killing their targets. With the LTCA I can also examine how homogeneous the response is within a population; are certain T cells more efficient at killing their target cells or do all cells in the population behave the same?”
Nanolive would like to thank David for taking the time to talk to us and for sharing some of his fascinating videos with us.
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