Journal: The American Journal of Pathology
Article Title: Retinal Vascular Abnormalities and Microglia Activation in Mice with Deficiency in Cytochrome P450 46A1–Mediated Cholesterol Removal
doi: 10.1016/j.ajpath.2018.10.013
Figure Lengend Snippet: Macrophage/microglia activation in the Cyp46a1−/− retina. A and B: Representative stains of retinal flat mounts for F4/80 (red; A) and retinal cross sections for Iba1 (red; B); nuclei were stained with Hoechst (blue) and DAPI (blue), respectively. C: Enlargement of the boxed area in B showing wild-type retina with Iba1-positive cells. n = 3 (A and B). Scale bars = 50 μm (B). Original magnification, ×200 (A). 2°, Control stain using secondary antibody only; GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; NI, control stain using nonimmune serum; ONL, outer nuclear layer; OPL, outer plexiform layer; RPE, retinal pigment epithelium.
Article Snippet: Retinal flat mounts were washed for 1 hour at 4°C with four changes of PBS and then mounted with either ProLong Gold without DAPI (for F4/80 staining) or VECTASHIELD mounting media with DAPI (H-1200; Vector Laboratories, Inc., Burlingame, CA) (for CYP46A1 and CD31 staining).
Techniques: Activation Assay, Staining