Preparation of retinal vessels for scanning electron microscopic examination using microdissection by ultrasonication

A method of microdissection by ultrasonication was used to prepare blood vessels of the retina for scanning electron microscopic examination. Eye cups were treated with 2% OsO4 that contained 2% CHAPS or 1% saponin (16 hrs.), dehydrated to 100% acetone, sonicated at 80 kHz (40 min.), and further processed by conventional means. This treatment resulted in the separation of the outer and inner retina at the level of blood vessels in the outer plexiform layer. The overall pattern of the blood vessels was maintained. Venules were distinguished from capillaries. This technique may provide information that is not possible to obtain with corrosion casts or trypsin digests.