THE ULTRASTRUCTURAL LOCALIZATION OF CYTOCHROME c-OXIDASE COMPLEX IN A LIPID-RETAINING, GLUTARALDEHYDE-UREA EMBEDMENT

Kidney tissue, incubated in a phosphate-sucrose buffer with diaminobenzidine (DAB), subsequently was embedded in polymerized glutaraldehyde-urea (Pease and Peterson, 1972). The highly polar character of this embedment retains lipids in ultrathin sections and thus permits a precise localization of reaction products in relation to cytomembranes. Furthermore, since conventional organic solvents are not used during processing, it is thought that oxidized DAB polymers certainly remain in place. Their density can be enhanced by exposing mounted sections to OsO 4 vapor, rather than by en bloc staining. DAB oxidation takes place only in the compartment between the inner and outer mitochondrial membranes. When aldehyde-fixed tissue is incubated, the deposits are largely limited to the intracristal spaces, whereas when fresh tissue is incubated, the entire compartment is uniformly filled. Morphologic features of fresh, unfixed tissue are stabilized by ethylene glycol and so survive incubation best when about 30% of this substance is added to the medium.