Bz x700 light and fluorescent microscope

A freezing sliding microtome (Microm HM 450, Thermo Fisher Scientific) was used to generate coronal sections (40 μm) spanning the infarct (Bregma 0.74 mm – 2.54 mm). Immunostaining was performed on free-floating brain sections using standard techniques (Doyle et al., 2015 (link); Nguyen et al., 2016 (link); Zbesko et al., 2018 (link)). The following primary antibodies were used: B220/CD45R (BD Biosciences, Cat. No. 553085, RRID: AB394615), CD3ε (BD Biosciences, Cat. No. 550277, RRID: AB393573), immunoglobulin A (IgA; BioLegend, Cat. No. 407004, RRID: AB315079), and CD138 (Syndecan-1;BioLegend, Cat. No. 142514, RRID: AB2562198). Sections were then labeled with the appropriate secondary antibody in conjunction with ABC Vector Elite and 3,3′-diaminobenzidine kits (Vector Laboratories) for visualization. For fluorescence imaging, sections were incubated in appropriate Alexa Fluor secondary antibodies (Thermo Fisher Scientific). Sections were imaged using a digital Keyence BZ-X700 light and fluorescent microscope or a Leica SP5-II laser scanning confocal microscope.