Protocol to characterize mouse dural mast cells by flow cytometry and immunofluorescence

Mast cells, which constitute tissue-resident immune cells, are distributed in the dural meninges. Here, we provide procedural guidelines for investigating mouse dural mast cells using two techniques. First, we outline the procedures for dural tissue dissection, single-cell isolation, and subsequent surface staining for mast cell identification via flow cytometry. We then describe the techniques employed for whole dura tissue staining to visualize mast cells using confocal and slide scanning microscopy, followed by analysis using Nikon’s NIS-Elements Advanced Research software. For complete details on the use and execution of this protocol, please refer to Lin et al.1 [Display omitted] •Isolation and characterization of mouse dural mast cells with flow cytometry•Whole dura harvest and staining to visualize mast cells with confocal microscopy•Analysis of dural mast cell images with NIS-Elements Advanced Research software Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Mast cells, which constitute tissue-resident immune cells, are distributed in the dural meninges. Here, we provide procedural guidelines for investigating mouse dural mast cells using two techniques. First, we outline the procedures for dural tissue dissection, single-cell isolation, and subsequent surface staining for mast cell identification via flow cytometry. We then describe the techniques employed for whole dura tissue staining to visualize mast cells using confocal and slide scanning microscopy, followed by analysis using Nikon’s NIS-Elements Advanced Research software.