I need to study these cells in whole blood samples. I normally use the protocol "Lyse-no-wash". It could be used to study these cells, too?
Hy Massimo. If I am on time to help... Neutrophils: CD68, CD15; Eosinophils: Siglec-8 (the human counterpart of Siglec-F); Dendritic cells: this is tricky... there are several populations of DCs in blood. I worked a lot with this kind of cells and our approach usually was to exclude other lineages (CD3, CD56, CD14, CD19 in the same colour; there are mixtures of these mAbs usually called Lineage exclusion kits) use HLA-DR and select those with high expression. Then you should use CD33 and CD123 and CD16. You can find 3 populations of these cells usually... If you want I can send you some papers with the procedure.
Hy Massimo. If I am on time to help... Neutrophils: CD68, CD15; Eosinophils: Siglec-8 (the human counterpart of Siglec-F); Dendritic cells: this is tricky... there are several populations of DCs in blood. I worked a lot with this kind of cells and our approach usually was to exclude other lineages (CD3, CD56, CD14, CD19 in the same colour; there are mixtures of these mAbs usually called Lineage exclusion kits) use HLA-DR and select those with high expression. Then you should use CD33 and CD123 and CD16. You can find 3 populations of these cells usually... If you want I can send you some papers with the procedure.
Hi Jose, your expertise is really helpful for me! if you can please send me your papers! Thank you very much!
Neutrophils and eosinophils are both in the granulocyte gate, neutrophils are CD16 positive and eosinophils are CD16 negative. CD15 is expressed both on eosinophils and neutrophils. You can even distinguish eosinophils in unstained samples by their autofluorescence
The best marker for Eosinofil and neutrophil differentiation is CD16 and also Eosinophils are more granuler FS/CD45 help to differentiate.
Hi Jose, would you mind sending your papers to me ? thanks in advance
Probably I am so late. But I hope this information is helpful...
- pDCs: CD123+bright, HLADR+. In addition, there are some specific markers that can also be analyzed: CD303+, CD304+, CD141+.
- mDCs: CD33+++, CD14/CD16-, HLA-DR+ (can also be helpful: CD2+, CD11c+, CD300e+). You can divide these in mDC1 (CD1c+, CD1a+) and mDC2 (CD141+).
- Neutrophils and eosinophils can easiliy be differentiated by the expression of CD16 (negative in eosinophils) or CD157 (negative in eosinophils).
- Basophils: CD123+ bright, HLA-DR-, CD33++, CD22+ weak (can also be helpful: CD203+, CD38+ bright)
Thanks Xavier, I have a question about Basophils. What do you think about SS(low)CCR3+ or CD3-CRTH2+? CD203c+ is useful only for activated basophils. It's right?
WBW
Massimo
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