Nisarg Soni,

We had decent results for mouse and rat blood using Ficoll-Paque at 1.084 g/m, but it primarily yields lymphocytes, but that was the population we were trying to collect.  In humans or animals, we were able to use heparinized or EDTA blood as our sample source.  

As for PBS versus balanced salts solution, we primarily used PBS.  Depending on your final use of the wbc's, balanced salts solution might have benefits for some experimental needs but once separated could be resuspended in medium or your prefered balanced salts solution.  We noticed no difference in our own applications between PBS and Hanks Balanced Salt Solution (HBSS) used in the initial collection and wash steps.   Once washes were completed, the cells would be resuspended in medium or HBSS.

Paul Taylor

Hi Nisarg, heparin, EDTA collected blood does not affect ficoll. You just have to dilute the blood 1:1 to reduce density. It does not matter if you use PBS or saline to do that.

Just to expand on Paul's answer, ficoll will give you a band of PBMC's, but granulocytes will pellet with the RBCs

If you want to isolate all WBCs, then a RBC hemolysis step followed by centrifugation might be what you are looking for. 

Agreed, Steingrimur Stefansson is very true on the Ficoll.  I never was concerned with granulocytes for my particular application, but a few of the individuals in our group did RBC lysis for isolation of all WBCs.  The method worked well on both human and animal samples.  We verified populations using flow cytometry and had very good results.   Good luck in your isolation.

For MNC, in our hands, especially in human neonatal blood or bone marrow samples, first Ficoll and 2nd short and non aggressive  lysis (sometimes only H2O) worked very well, also for getting rid of nucleated red blood cells.

For WBC (inlcuding granulocytes) we do in samples with high platelet concentration at a first step a platelet wash including plasma removal, and secondly an erythrocyte lysis (NH4 / EDTA/ HEPES buffer) with  -depending on  the application- subsequent lysis buffer wash.