I agree with Julie that CD11b and Gr-1 are the best markers currently available to resolve the phagocytic populations in mouse peripheral blood. The situation may be more complex. Attached is a contour plot of 250,000 CD45-PE/Cy5 bright leukocytes stained with Gr-1-FITC and CD11b-PE using a no-lyse/no wash protocol (1). Depending on ones degree of bravery, there are 5-7 populations that can be identified. Populations A/B (Gr-1 & CD11b bright) are granulocytes, I have noted that the proportion in B is larger after erythrocyte lysis. Pops C & D (Gr-1 neg, CD11b medium & high) are monocytes and based on (2), likely a subpopulation of B-cells although I do not know which ones are which. Pop E (Gr-1 low, CD11b neg) is probably a T-cell subpopulation (2). Population F (Gr-1 & CD11b low) is unknown but the peak increases as cells with lower CD45 expression are included in the gate. Pop G, (CD11b & Gr-1 neg) is the bulk of T & B lymphocytes in peripheral blood. -Jim Weaver Refs (1) Serial Phenotypic Analysis of Mouse Peripheral Blood Leukocytes. James L. Weaver, Dennis D. Broud, Katherine McKinnon, and Dori R. Germolec. Toxicol. Mech. Methods 12 95-118 (2002). (2) Flow cytometric identification of murine neutrophils and monocytes. Eric Lagasse & Irving Weissman. J. Immunol. Meth. 197 139-150 (1996). * -----Original Message----- * From: Oughton, Julie [mailto:julie.oughton@orst.edu] * Sent: Wednesday, September 11, 2002 6:44 PM * To: Cytometry Mailing List * Subject: RE: markers to distinguish mouse monocytes * * * * Oops! * * I misspoke in my message re: the best markers to identify * monocytes in mouse * blood. I still stand by CD11b and Gr-1 as the best markers to resolve * monocytes from granulocytes. As you will see from the * histograms illustrated * in the attached word document, both monocytes and * granulocytes express CD11b * and Gr-1. However, granulocytes express high levels of Gr-1 * while monocytes * express low levels of Gr-1 [I had previously said that only * granulocytes * express Gr-1, my mistake]. I used color eventing to show the * profiles of * these two populations. * * Sorry for the mistake! * * Julie Oughton * Oregon State University * * * * <<CD11b Gr-1.doc>> *
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