We thought we would write a blog about using our flow cytometry software VenturiOne to look at one of the OMIP (Optimised Multicolour Immunofluorescence Panels) using the files from the flow repository and gating strategy which is detailed in the paper and supplementary information (links below)
We chose OMIP 30 : Characterisation of human T cell subsets via surface markers. This panel uses 17 fluorochromes and a live/dead stain on PBMC’s (peripheral blood mononuclear cells) to differentiate the different lineages of T cells present, focussing on the well described CD4+ T helper subsets and the Tregs.
Recreating the gating strategy was a little more intensive than I first thought, using VenturiOne to create the plots was easy enough, but the more taxing part was after getting around 20 gates in and having to deal with gates diverging at each step and forming their own consecutive gating branch and double and triple checking I had not missed plots and gates along the way!
Thankfully, the Hierarchy view helped enormously with this, allowing me to check at a glance that the parent gate was correct, and I had not missed out anything.
The outcome is above
When so many parameters are being compared there is almost too much information to take in and this drives home the message that there is increasing need for an easier way to look at multiparameter data and it would be beneficial if it was in a predictable and recognisable format.
The Citation and the data can be found at the links below, feel free to download a free trial of VenturiOne to have a go for yourself and let me know how you get on creating this gating strategy.
The paper and supplementary information can be found
and the data is available on the flow repository website