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.
Acton
Anthony
A
Department of Biochemistry and Molecular Biology, Center for Medical Genomics, Indiana University School of Medicine, Indianapolis, Indiana, USA.
Harvey
Tia
T
Grow
Matthew W
MW
D000818:Animals; D004625:Embryo, Nonmammalian / Q000378:metabolism; D019286:Endopeptidase K; D005557:Formaldehyde; D017930:Genes, Reporter; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D011108:Polymers; D016707:Tissue Fixation / Q000191:economics* / Q000379:methods*; D014157:Transcription Factors / Q000378:metabolism; D019210:Troponin I / Q000378:metabolism; D029867:Xenopus Proteins / Q000378:metabolism; D014982:Xenopus laevis
D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C092720:NKX2-5 protein, Xenopus; D011108:Polymers; D014157:Transcription Factors; D019210:Troponin I; D029867:Xenopus Proteins; D005557:Formaldehyde; D019286:Endopeptidase K; C003043:paraform