Confocal laser scanning microscopy of retinal rod outer segment intact disks: a new labeling technique

摘要

Vertebrate retinal rod outer segment disks house\udthe proteins involved in the phototransduction cascade\udthat converts light into neuronal signal. We develop a\udtechnique for the immunofluorescent labeling of osmotically\udintact isolated rod outer segment disks for confocal\udlaser scanning microscopy imaging. Osmotically intact\udFicoll-flotation isolated bovine disks are directly labeled\udwith antibodies in solution. For the first time, osmotically\udintact single disks can be visualized. Thus, imaging of purified\uddisks, based on advanced optical techniques, may\udserve as a powerful complement to other methods in studies\udon phototransduction. In fact, even though much is\udknown about the rod outer segment photoresponse, some\udunanswered questions remain, particularly about ATP supply,\udlight adaptation, and morphogenesis.