[Cytometry] Human monocyte preparation for flow analysis

Patrick-Melin, Amy patrica2 at ccf.org
Fri Feb 20 10:12:17 EST 2009


I am a graduate student that is new to flow cytometry.  I am working with human monocytes and looking at
surface expression of TLR2 and TLR4 via flow.    
 
I am currently working with human primary monocytes in cell culture and having difficulty removing the cells
from the plates.   Can anyone tell me whether the cells are damaged with cell scraping?  Does anyone have
a protocol for removing monocytes from polystryene cell culture wells?  We currently isolate MNCS with
a Histopaque method and confluency is close to 100% after 24 hr. incubation, however, we losing the cells
somewhere in the preparation for flow.
 
Any guidance would be deeply appreciated.  
 
Thank you in advance,
 
AJPM  
 
 

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