I need to differentiate macrophages from monocytes and other cells. Can I do that by flow cytometry, and what markers and antibodies can I use?
D Pilling and his group did a similar work identifying macrophages using CD makers. It may be helpful
Macrophages can be identified by specific expression of a number of proteins including CD14, CD11b, F4/80 (mice)/EMR1 (human), CD68 and MAC-1/MAC-3 by flow cytometry
Macrophages can be identified markers such as CD14, CD11b, F4/80 (mice)/EMR1 (human), CD68 and MAC-1/MAC-3. One of the more important receptors in macrophage biology is PECAM-1 (CD31),
In body fluids I've sometimes had even small percentages of macrophages interfere with staining by "mopping up" antibodies with Fc receptor. As a matter of protocol I now FcR block body fluids before staining. One can use CD64 instead. In bone marrow and tissues I seldom see a significant population.
It's pretty hard to miss them when they're present. The SSC is high, and your monocyte markers such as CD14 and CD64 will separate the population.
By the way, I was recently looking into flow cytometry for Langerhans cell histiocytosis. You might find this article interesting: http://bloodjournal.hematologylibrary.org/content/97/5/1241.long .
The article below may be helpful as they are looking into macrophage differentiation following stimulation and using flow, confocal microscopy, apoptosis assay and cytokine studies. I guess it will give a better understanding o the surface markers and the differention of macrophages from monocytes as discussed nicely and demarcated.
Citation: Daigneault M, Preston JA, Marriott HM, Whyte MKB, Dockrell DH (2010) The Identification of Markers of Macrophage Differentiation in PMA-Stimulated THP-1 Cells and Monocyte-Derived Macrophages. PLoS ONE 5(1): e8668. doi:10.1371/journal.pone.0008668
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0008668#pone-0008668-t001
- Badges
- Science topic