Mechanical isolation of neonatal and adult mouse dura leukocytes for flow cytometry analysis

The meninges, consisting of the pia, arachnoid, and dura layers, provide immunosurveillance of the central nervous system with both innate and adaptive immune cells. Here we present an optimized protocol for isolating dura leukocytes from neonatal and adult mice. We describe steps for harvesting the skull cap, extracting the dura mater, mechanical isolation of dura leukocytes, and flow cytometry analysis. Unlike the time-consuming enzymatic digestion isolation which makes dura hematopoietic stem cells (HSCs) undetectable, this rapid and simplified technique permits dura HSC identification. For complete details on the use and execution of this protocol, please refer to Niu et al. (2022).¹.