Has anybody tried reducing PE-Cy5 or Cy5 binding to monocytes (via CD64) by blocking with unlabeled anti-CD64? Probably an expensive solution, but no more so than an irrelevant Cy5 conjugate. Also, since in my limited experience (hacking around at the Maine flow course a few years ago with Alan Waggoner and Carl Stewart) Cy3 as well as Cy5 conjugates bind to monocytes, I would guess that a Cy7 conjugate would as well; unless you are using APC-Cy7 or PE-Cy7, this should reduce binding of the Cy5 or PE-Cy5 conjugate without spectral interference. -Howard
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