Figure 4.

Neutral lipid stains in BrM wholemounts. Outer macula, RPE removed. Arrowheads, intercapillary pillars of BrM; asterisk, BrM over choriocapillaris. (A) In unstained section viewed with DIC, intercapillary pillars were hardly visible. Shown in lower right is remaining choroidal pigment. (B) Section stained with Oil Red O viewed with bright-field microscopy with light-red coloration of BrM and intense outlining of the intercapillary pillars. (C) In dark-field mode, weak fluorescence of Oil Red O of intercapillary pillars only. Wholemounts stained with Nile Red (D) and BODIPY 493/503 (E) show comparable patterns, with strong background fluorescence from the choroid. Intercapillary pillars are visible only if images are regularly enhanced by image-processing software (contrast/brightness). (F) UC stained with filipin. Upper right, an island of RPE was left for better orientation. Weak, inconsistent signal from BrM, major interference by choroidal connective tissue background. (G) EC: filipin regularly stained a granular pattern with increased intensity in intercapillary pillars. (H) Wholemount preparation imaged with same filter (UV) and image-capture settings as in F and G. Bar = 40 μm.