Microglia Morphology Analysis in Retinal Sections

Three 40× field views were captured from the corresponding grafted areas of three 15-μm thick retinal sections per eye in the C-Kit⁺ cells group and the sham group using a Leica confocal imaging system with 1-µm z-steps. Maximum intensity projections (MIPs) were processed. Subsequently, the morphology of the microglia in the sections was analyzed using a grid system to determine the number of grid-crossing points per individual cell (from at least three different eyes per group) as previously reported^{34 (link),35 (link)}. The number of grid-crossing points per cell and the relative frequency of crossing were analyzed. The numbers of Iba1⁺ cells and Iba1⁺/CD68⁺ cells were quantified by counting the cells in sections of retinas (n = 5 eyes per group).