June 10, 2011  |  Flow Cytometry  |  By  |  1 Comment

Working with Accuri Data

Cytobank users have uploaded and analyzed data collected from more than 30 different flow cytometer models, so chances are that Cytobank can handle your data! In a recent post, we featured the ability of Cytobank to facilitate the mining of data from large datasets generated by the DVS Sciences CyTOF. This time, we will walk you through analysis of data collected on the Accuri cytometers using their CFlow software.

Image plots from Accuri CFlow

Accuri provided us with a set of sample files demonstrating the collection of data from cells stained with a PE-anti-CD4 antibody, and we’ll use this as an example. You can see from their CFlow software analysis that they achieve separation of and gate on the lymphocyte population (P1, first panel), and further separate CD4+ from CD4- cells (second two panels). We’ll show you how to do the same in Cytobank!

Configuring Plot Scales on Cytobank

After collecting your data in CFlow, export the data in the FCS format under the File menu (watch a video demonstrating this). Next, upload these FCS files to Cytobank by creating a new experiment. If your data look condensed toward the plot origin, you can easily spread them out by adjusting the Plot Scales ranges. Under Plot Scales on the Illustration page, change the FCS-A maximum value to 2,300,000 and the SSC-A maximum value to 1,000,000 if these scatter channels were collected in linear mode. From here, all annotation and analysis is accomplished as described in our tutorials.

Scatter gating (left) and CD4-PE gating (right) in Cytobank

If you have a question about whether data from your cytometer is compatible with Cytobank, send us an email! If your cytometer isn’t part of the more than 30 cytometers with which we have experience, send us a sample FCS file and we’d be happy to test it!

– Angela