Figure 6: Accuracy of immunofluorescent staining validated via confocal microscopy. LqN2/isopentane cryosections of skeletal muscle were fixed in acetone at room temperature and stained as before and visualized by scanning confocal microscopy. Images were acquired using sequential scans for hF.IX (red), CD8 (green), and DAPI (blue). (A) Low and (B) high magnification views showing both hF.IX expressing cells (red) and nonhF. IX expressing cells (*) with infiltrating CD8+ (green) lymphocytes. (C) Region of non-hF.IX expressing muscle tissue with DIC overlay inorder to visualize negatively stained cells. (D) Lower magnification of (B) with DIC overlay in order to visualized the non-expressing cells.