Wafer embedding: specimen selction in electron microscopic cytochemistry with osmiophilic polymers.
A new wafer embedding procedure is described that permits light microscopic screening of embedded tissue prior to ultrathin sectioning. It is particularly valuable when used on specimens obtained with an automatic sectioner and treated cytochemically to obtain visible intermediate or visible and electron opaque final reaction products. Aldehyde-fixed tissues are cut into sections with an automatic sectioner, incubated cytochemically osmication if required, then embedded in epoxy resin between fluorocarbon coverglasses which are supported by a platform specially designed for this purpose. The resultant wafter, less than 0.2 mm thick, is examined by light microscopy for optimal areas of cytochemical reaction and desirable structural features. Such areas are cut out and glued to blank blocks with fast curing cyanoacrylate cement for subsequent ultrathin sectioning. The usefulness of this technique is demonstrated by the location of: (1) esterase-positive lysosomes in kidney and trigeminal ganglia; (2) palatal sensory endings stained for acetylcholinesterase; and (3) phagosomes arising from the resorption of horseradish peroxidase tracer by the cuboidal parietal epithelial cells of Bowman's capsule in the male mouse.
D000110:Acetylcholinesterase / Q000032:analysis*; D000818:Animals; D004950:Esterases / Q000032:analysis*; D005069:Evaluation Studies as Topic; D005724:Ganglia / Q000201:enzymology / Q000648:ultrastructure; D006735:Horseradish Peroxidase / Q000032:analysis*; D007668:Kidney / Q000201:enzymology / Q000648:ultrastructure; D008247:Lysosomes / Q000201:enzymology / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron / Q000379:methods; D010544:Peroxidases / Q000032:analysis*
D006735:Horseradish Peroxidase; D010544:Peroxidases; D004950:Esterases; D000110:Acetylcholinesterase