Fig. 1

Laser CNV model and flow cytometry gating strategy for mononuclear phagocytes. IBA1 (a), ICAM-2 (b), and merged (c) images of a flat-mounted choroidal lesion on day 3. d Schematic of experimental design. Red arrow indicates that the representative plots in e, and j–m are from day 3 after laser injury. e Initial gating strategy demonstrating identification of singlet cells (left), delineation of live cells with removal of debris and count beads (middle left), gating of CD45⁺ cells (middle right), and selection of CD11b⁺, Lin^- cells (Lin = CD4, CD8, B220, NK1.1, SiglecF, Ly6G). f–m Gating strategy for identification of monocytes, dendritic cells (DCs), microglia, MHCII⁻, MHCII⁺CD11c⁻, and MHCII⁺CD11c⁺ macrophages from unlasered (f–i) and laser treated (j–m) mice. f, j Separation of CD45^High from CD45^Dim cells. h, l Delineation of microglia as CD45^DimCD64⁺MHCII^Low cells. g, k CD45^HighCD64⁺MHCII⁻ macrophages (MHCII⁻ Macs) are increased with laser. i, m Increased CD45^HighCD64⁺MHCII⁺CD11c⁻ (CD11c⁻ Macs) and CD45^HighCD64⁺MHCII⁺CD11c⁺ macrophages (CD11c⁺ Macs) with laser treatment