I'm trying to do flow cytometric analysis of the inflammatory cell infiltrate in the arthritic joints in mice. I've been targeting the ankle and "wrist" joints. I've tried to wing it (flushing with 1 mL PBS/2 mM EDTA/10% FBS using a 25G needle) but the variability from mouse to mouse is very high. I haven't found an article that describes the precise methodology for flushing the joints for flow or cytokine analysis. Does anyone have a protocol they trust?
With thanks,
Michael
Hey Mike :) What you describe is exactly what I did earlier - however I believe I went with 3 separate flushes rather than just one - typically helps to get everything out when you flush more than once (a bit like a BAL). I also remember using joints which were cut out right before and after the ankle so I had access to the joint quite easily and the liquid that came out was pretty clear. It was a bit messy but I got plenty of inflammatory cells and not much blood. The results we got then were pretty consistent. I'll send you the PDF by email.
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