neutrophil isolation

mark mark.haynes at mail.tju.edu
Thu Feb 4 19:48:41 EST 1999


Eric Park wrote:
> 
> I'm encountering some problems when I try to isolate neutrophils from my
> blood.  I'm using a Ficoll-Hypaque solution when I centrifuge the blood.
> This technique works great for several other people but not for me.  A lot
> of RBCs are remaining in the neutrophil layer and not settling down to the
> bottom of the tube.  Does anyone know why this is happening to my blood?

First the others had good answers vis a vis Temp and dilution etc but i was 
under the impression that f/h centrifugation results in mononuclear cells 
and most granular/polymorphonuclear cells are not inthe interface which is 
what i thought a neut was, its a monocyte?

there is definately a problem if blood is cooled don't do that.  also some 
people's blood does act slighly differently in the technique.  for example, 
sickle cell or thalassemia RBC's don't work very well in f/h centrifugation.
But be the vampire and use all other possible people-- give the grads student 
a $20 (covered by you r dept of course) and you probably won't have a problem 
gettin blood
good luck




> Personal info:
> Blood type is B+ Rh+
> Asian male 22 years old
> 
> Insights would be helpful since this is a mildly annoying problem in my
> research by begging for volunteers instead of tapping my own blood.



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