Ronald Rabin wrote: > Lately, the flux in response to injection of buffer has been huge, making it > impossible to accumulate any new data. Putting the cells on the cytometer, > taking them off, and then putting them back on even causes a calcium flux, > so the cells are responsive to changes in pressure, agitation or > perturbation, but only after initially being placed on the cytometer. I don't know the exact construction of the Time Zero Module, but from what you describe I suspect a bacterial contamination in the tubings. Bacterial LPS is a very potent stimulator of macrophage functions, and also leads to the release of intracellular calcium. As in a traditional fluorimeter the cells "behave well", something in the sorter causes the problems. It may be worthwile to change the injection tubings and check the calcium flux. Noteworthy, human lymphocytes would not be affected by such a contamination. Jens
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