I used miltenyi whole blood CD3 micro beads to separate the T cells from whole human blood using the automacs seperator. Usually after the separation is done by the machine, there is a clear T cell isolate. Yesterday I ran the machine twice and the isolate had brown coloured sediments at the bottom both the times—probably micro beads. I have been using the same settings for a long period with no issues. Has anyone encountered this before and can someone please help troubleshoot? is this an issue with the beads or the column or something with the setting?
I am not sure why this happened, but you can double check the micro beads may be this turbidity from it or from the buffer. Did you check the cells under the microscope? what did you seen? maybe this will give you any idea to solve this issue. Good luck.
Thanks Fahad!
I vortexed the beads and the sediments were far less. Probably it could be related to the beads. I ran the buffer one with possel (also prepared a new one), it had no sediment. So unlikely related to buffer turbidity, I guess?
I was able to see the T cells under the cell counter. However it did have big chunks of beads as well.
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