I have performed a PMNL isolation from human whole blood (dextran sedimentation, ficoll-paque density gradient centrifugation, red blood cell (RBC) lysis).
When I have checked the percentages of netrophils in the whole blood, the percentage was normal to me (2,7%), but when I checked the percentage of eosinophils after the PMNL isolation, the percentage is too high (25%).
Does anyone had an explanation for that? When working with Buffy Coats, it doen't happen.
Thank you in advance for your help.
The donor might had worm infection or allergies that might resulted in high Eosinophils
Hi Llobet,
When you checked the percentage of eosinophils in whole blood, the number of eosinophil is calculated among all kinds of WBC presenting in the blood (neut, baso, lymp and eos). After isolating by gradient centriguation and sedimentation, you just only get Neut and Eos, The percentage of eos is certainly increased.
Another possible reason is that Neut could die during the process of isolation, this causes the reduction of Neut count and therefore increases the percentage of Eos.
I hope this can explain your concern.
Warm regards.
Thank you Le-Duy Pham for your answer. YOu are right. The problem was due to the red cell lysis. In that step I lost a significant part of neutrophils and that was the reason I further had this high percentage of eosinophyls.
Thank to anyone who tried to help me.
Regards.
Exaclty - eosinophils are more resistant to RBC lysis than neutrophils. This is well known and part of many lab manuals, but unfortunately - to my knowledge - unpublished.
i think the donor might had worm infection or allergies that resulted in high Eosinophils, such individual need to be check for parasitic infection. the number of eosinophil can be calculated among all kinds of White blood cells present in the blood (neut, baso, lymp and eos).