Lsr fortessa series

The following mAbs were used for cell surface staining: anti-CD45-PerCpCy5.5, anti-Ly6G(1A8)-PE or FITC, anti-Ly6C-APC or FITC (all from BD Bioscience), anti-CD11b-Pacific blue, anti-CD11c-PECy7, anti-CD4-AF700, anti-IL-4-FITC, and anti-IFN-γ-PECy7 (all from eBioscience); anti-CD3-PE or APC and anti-IL-10-PE (Biolegend); and anti-CD8-APC (Invitrogen). For the exclusion of dead cells, we used the Live/Dead fixable Aqua Dead Cell Stain Kit (Invitrogen). L. major mCherry was detected using Texas red as a positive control, L. mexicana DsRed and L. major Sd-RFP were detected using PE as a positive control. Fluorescent parasites and stained murine cells were acquired using a flow cytometry analyzer, either BD LSRII or BD LSR-Fortessa Series (Beckton Dickinson) machines, the data was acquired with the DIVA software and the results were analyzed with FlowJo 10 software.

Surface activation markers and cell viability markers in blood derived human neutrophils were analyzed using Flow Cytometry analyzer of either the BD LSR II or the BD LSR Fortessa series (Becton Dickinson) and analyzed with FlowJo software (Tree Star). The following antibodies were used: Anti-human: CD15-APC, CD62L-PErCP-efluor710, CD66b-PE-Cy7. Anti-mouse: Ly6G(1A8)-APC-Cy7, CD62L-PE, CD11b-PE-Cy7 (all from e-Bioscience). Cell viability: Live/Dead fixable Aqua Dead Cell Stain Kit (Invitrogen).