Abstract
Lowicryl resins are well suited for electron microscope immunocytochemistry of formaldehyde or formaldehyde/glutaraldehyde fixed tissues. In immunogold electron microscopy, section staining is important in order to determine the precise relationship between colloidal gold particles and tissue fine structure. Freeze substitution and Lowicryl HM20 embedding combine good preservation of cell ultrastructure and antigenicity. Chemicals used during dehydration and embedding resins are toxic and should be handled with adequate safety precautions. Cryosectioning and freeze substitution followed by Lowicryl embedding are alternative procedures for immunoelectron microscopy. Immunolabeling of sections from freeze-substituted and Lowicryl-embedded tissues usually provide a very high precision in probe localization. Ultrathin sections of aldehyde fixed and Lowicryl HM20 or K4M-embedded tissue can be used reproducibly and conveniently for immunocytochemical detection of many antigens. High background staining can, in some cases, be prevented by increasing the sodium chloride concentration or the pH in the incubation solution for the primary antibody and/or the secondary gold conjugate.
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