How can we separate young and old mouse neutrophils? What are these cell's significant characteristic? What kind of techniques do you use to separate these cells?
A little off topic. In smears I use NASDCL staining (naphtol-AS-D-chloroacetate esterase). The pattern changes as well as the intensity.
Most of the time I only do morphology on a a Diff-Quick or similar staining.
If you need I can send you some pictures. For simplicity the band forms have a donut- like nucleus, the more mature have a croissant-like one and the matures, have segmented one, like multiple small lobes. Also the chromatin "condenses" between band and mature forms. The granularity increases from band to mature and the granules are tiny and slightly reddish (azurophilic).
This is from the morphology point of view but I assume you want to do FACS, right?.
Cell. 2013 May 23;153(5):1025-35. doi: 10.1016/j.cell.2013.04.040.
Rhythmic modulation of the hematopoietic niche through neutrophil clearance.
Casanova-Acebes M, Pitaval C, Weiss LA, Nombela-Arrieta C, Chèvre R, A-González N, Kunisaki Y, Zhang D, van Rooijen N, Silberstein LE, Weber C, Nagasawa T, Frenette PS, Castrillo A, Hidalgo A.
Casanova-Acebes et al's paper will be very useful for that. Main markers for FACS would be CD62L, CXCR4 & CD11b. On how to separate them, you may try sorting based on fluorescence levels for the previous markers.
Article Rhythmic Modulation of the Hematopoietic Niche through Neutr...
Thank you İvan for yor useful suggestions. For understand my morphology study, surface markers should be necessary.
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