Epi-polarization and incident light microscopy readily resolve an autoradiographic or heavy metal label from an obscuring background or second label.
Difficulty encountered in resolving grains of exposed photographic emulsion in autoradiographs of the densely melanized retinal pigment epithelium was solved by using epi-polarized or incident light microscopy. The apparatus used included a metallurgical illuminator specifically designed for epi-polarization microscopy or, as a less expensive but only slightly less effective alternative, a modified fluorescence illuminator. The black melanin granules absorb incident light (as they do in vivo) while the silver grains reflect it producing a "darkfield-like" representation. Brightfield and darkfield-like images can be alternated easily and quickly, or both can be viewed simultaneously. Epi-polarization microscopy has wider application in resolving a reflective label over any opaque background staining or dark second label.
Rapaport
D H
DH
Department of Anatomy, University of Sydney, NSW, Australia.
Herman
K G
KG
LaVail
M M
MM
D000818:Animals; D001345:Autoradiography / Q000379:methods*; D006651:Histocytochemistry; D008027:Light; D008670:Metals / Q000032:analysis*; D008853:Microscopy / Q000379:methods*; D008859:Microscopy, Polarization / Q000379:methods*; D009893:Opossums / Q000033:anatomy & histology*; D010487:Periaqueductal Gray / Q000033:anatomy & histology; D012160:Retina / Q000033:anatomy & histology / Q000166:cytology; D012165:Retinal Ganglion Cells / Q000648:ultrastructure
D008670:Metals