A method permitting precise trimming of resin-embedded tissue for ultrathin sectioning in pre-embedding immunoelectronmicroscopy.
Due to the high complexity of the mammalian central nervous system, sampling of immunohistochemically processed brain tissues for electronmicroscopy requires an accurate and reliable technique. For this reason, the flat-embedding method, which allows light microscopical examination of tissue before sampling, is generally employed. Because of the osmification process, however, the tissue is blackish and opaque which hampers light microscopical selection of tissue areas of interest. We have found that tissue translucency is highly improved by an osmification process using an osmium tetroxide-ferrocyanide mixture. We describe a transilluminated chuck that enables visualization of immunostaining in specimens mounted on a trimming instrument, thus allowing for extremely precise sampling of the tissue.
Valentijn
J A
JA
Department of Animal Physiology, University of Nijmegen, The Netherlands.
van Daal
J H
JH
Jenks
B G
BG
van Abeelen
J H
JH
D000818:Animals; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure*; D004399:Dynorphins / Q000031:analogs & derivatives* / Q000378:metabolism; D004723:Endorphins / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D008854:Microscopy, Electron / Q000379:methods*; D008867:Microtomy / Q000379:methods*
D004723:Endorphins; D004399:Dynorphins; C036200:rimorphin