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CD4+ Yield in Whole Blood

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robs

Contributor

Posts: 40

Joined: Mon Dec 02, 2013 3:42 pm

Location: University of Connecticut

Post Thu Jul 02, 2015 6:17 pm

Re: CD4+ Yield in Whole Blood

Hi Chad,
Definitely let us know the results of the standard flow run. I remember having a lot of trouble with cell recovery using polystyrene 5mL FACS tubes, especially after fixation and during the water wash a good pellet never formed. I ended up moving most of my staining to polypropylene tubes and if possible used conicals since most of the cell loss seemed to come from wash steps.
Rob
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CRStevens

Master

Posts: 60

Joined: Thu Jul 17, 2014 5:07 pm

Post Mon Jul 06, 2015 3:28 pm

Re: CD4+ Yield in Whole Blood

Hi,

As an update I did a quick experiment trying a "hard fix" after my overnight fix/perm with Ir intercalator. I saw no difference in my yield of cells when I did a 1hr fix with 8% (fresh) PFA. I have heard from Fluidigm that they recommend doing a lysis first, wash, then stain before doing any fixation. Has anyone tried this and seen a difference?

-Chad
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