An Examination of Non-Formalin-Based Fixation Methods for Xenopus Embryos

Despite the growing availability of non-formalin-based fixatives, the vast majority of researchers in developmental biology continue to fix embryos and tissue in 4% paraformaldehyde. This fixation method has proven useful for both immunohistochemistry and in situ hybridization, yet working with paraformaldehyde has distinct disadvantages in its toxicity and the short shelf life of prepared solutions. In a search for viable alternative fixatives, we have evaluated two non-formalin-based commercial products, FineFIX (Milestone Microwave Laboratory System) and NOTOXhisto (Scientific Device Laboratory). These products were tested side-by-side with a commonly used 4% paraformaldehyde solution (MEMPFA) on Xenopus laevis embryos and assayed using whole mount immunohistochemistry and whole mount in situ hybridization. The results indicate that NOTOXhisto can be used as a substitute for MEMPFA in both tested Xenopus protocols with no loss of sensitivity or tissue morphology. Developmental Dynamics 233: 1464 –1469, 2005. © 2005 Wiley-Liss, Inc.