Increasing membrane contrast by means of imidazole-osmium post-fixation as exemplified by skeletal muscle fiber.
Until now, the interpretation of findings derived from investigations on membrane structures (T tubules, sarcoplasmic reticulum, the Golgi apparatus) in thick sections of mammalian muscle tissue has been limited in TEM due to the lack of sharp resolution of the membrane contours. This article shows how the imidazol-osmium post-fixation of tissue blocks can be used to achieve well-contrasted, sharply defined membrane contours. Therefore, unstained sections from imidazol-osmium post-fixed tissue can be examined immediately. But protein structures (e.g., ribosomes) remain uncontrasted with this technique. If needed, it is possible to visualize the protein structures by conventional section staining with uranyl acetate and lead citrate. This method is suitable for both ultrathin and thick sections (>150 nm).
Institute of Anatomy, Eberhard-Karls-University Tübingen, 72074 Tübingen, Germany. tvoigt@anatom.uni-tuebingen.de
Tilman
T
Voigt
Wolfgang
W
Dauber
Irina
I
Bensemann-Ryvkin
Xenia
X
Härtel
D000818:Animals; D005404:Fixatives / Q000737:chemistry; D007093:Imidazoles / Q000737:chemistry; D051379:Mice; D008867:Microtomy; D018482:Muscle, Skeletal / Q000648:ultrastructure*; D009992:Osmium / Q000737:chemistry; D016707:Tissue Fixation / Q000379:methods*
D005404:Fixatives; D007093:Imidazoles; D009992:Osmium