I measured microglia population by flow cytometry using a combination of CD45/CD11b,c antibodies in the healthy rat brain. Microglia population identified as CD11b,c+/CD45low cells represent 0.2% of total events.
I wonder if it is a correct value, considering the following protocol and the gating strategy:
Cell dissociation was performed with (DNAse/Collagenase D, 1 mg/ml) solution and myelin removal was performed by a 37% Percoll step. Then cells were stained with a mix of CD45/CD11b,c antibodies and 7AAD was added to discriminate dead cells.
Viable cells were gated from doublets discrimination and elimination of dead cells 7AAD+.
Positive populations were gated from no-labelled control cells and compensations settings were calculated from mono-labelled cells with median method.
Check if these papers help you and if the authors are here on RG:
J Neuroinflammation. 2017; 14: 101. Published online 2017 May 8.
Characteristics of primary rat microglia isolated from mixed cultures using two different methods
Li Lin,1,2 Rakhi Desai,2 Xiaoying Wang,2 Eng H. Lo,📷2 and Changhong Xing📷2
J Neurosci Methods. 2007 Aug 30; 164(2): 207–217.
Quantification of the rat spinal microglial response to peripheral nerve injury as revealed by immunohistochemical image analysis and flow cytometry
J. Blackbeard,a,⁎ K.P. O’Dea,a V.C.J. Wallace,a A. Segerdahl,a T. Pheby,a M. Takata,a M.J. Field,b and A.S.C. Ricea
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