Figure 5

Analysis of green fluorescent protein (GFP)-positive cells in the peri-infarct area by 2-photon-laser-microscopy (2PLSM). (A) Representative 2PLSM-image stacks of GFP⁺ microglia measured in a volume of 424.9 × 424.9 × 75 μm (x/y/z) before photothrombosis (PT) (pre) and in the peri-infarct area of a vehicle (d3-vh)- and an AMD3100 (d3-AMD)-treated mouse on day 3 after PT. Scale bars: 50 μm. (B) Images displayed in (A) after threshold modification according to Otsu [30] in order to measure soma sizes. (C) Quantitative analysis of GFP⁺ cells and their soma sizes in the peri-infarct area. Values are normalized to respective values of cell numbers and soma sizes before PT. Error bars represent standard error of the mean. Statistical analysis was performed by Student’s t test (*P < 0.05, AMD3100 versus vehicle treatment at the same time point; ^# P < 0.05, versus the same treatment at day 3). (D) Illustration of representative morphologies of GFP⁺ cells in a vehicle- and an AMD3100-treated mouse, respectively, before (pre), and on days 3, 7 and 14 after PT. Scale bars: 10 μm.