X-Message-Number: 12698 Date: Tue, 2 Nov 1999 20:20:11 -0800 (PST) From: Doug Skrecky <> Subject: inert dehydration with ethylene glycol Citations: 1-4 <1> Authors Stirling JW. Coleman M. Brennan J. Institution Department of Pathology, Flinders Medical Centre, Bedford Park, South Australia. Title The use of inert dehydration and glycol methacrylate embedding for immunogold localization of glomerular basement membrane components. Source Laboratory Investigation. 62(5):655-63, 1990 May. Abstract A protocol is described for the preparation of human pathology specimens without fixation, in order to perform immunocytochemistry at an ultrastructural level. Using the technique in conjunction with immunogold labeling, the basal lamina components type IV collagen, laminin, and heparan sulfate have been demonstrated in glomerular capillary loops in stored frozen human renal tissue. Tissue was thawed and immediately dehydrated with the inert cryoprotectant ethanediol (inert dehydration) followed by embedding in low-acid glycol methacrylate polymerized using the accelerator n,n-dimethylaniline. Tissue processed in this way retained superior antigenic activity when compared with tissue reprocessed from wax blocks and embedded in low-acid glycol methacrylate. Inert dehydration is a technique useful for the localization of processing sensitive epitopes in routine fresh or frozen archival pathologic material. Furthermore, high probe densities can be achieved without recourse to etching or enzyme treatments. <2> Authors Sedar AW. Title Electron microscopic demonstration of polysaccharides associated with acid-secreting cells of the stomach after "inert dehydration". Source Journal of Ultrastructure Research. 28(1):112-24, 1969 Jul. <3> Authors Koscianek-Malczewska H. Komender J. Ostrowski K. Title Investigation of the N and P losses in different media used for inert dehydration. Source Experientia. 23(4):279-80, 1967 Apr 15. <4> Authors Pease DC. Title Inert dehydration. A new physical method of tissue preparation. Source Medical & Biological Illustration. 16(2):89-97, 1966 Apr. Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=12698