title: Alcohol-based solutions for bovine testicular tissue fixation.
abstract: Tissue fixation, a central element in histotechnology, is currently performed with chemical compounds potentially harmful for human health and the environment. Therefore, alternative fixatives are being developed, including alcohol-based solutions. We evaluated several ethanol-based mixtures with additives to study fixative penetration rate, tissue volume changes, and morphologic effects in the bovine testis. Fixatives used were Bouin solution, 4% formaldehyde (F4), 70% ethanol (E70), E70 with 1.5% glycerol (E70G), E70 with 5% acetic acid (E70A), E70 with 1.5% glycerol and 5% acetic acid (E70AG), and E70 with 1.5% glycerol, 5% acetic acid, and 1% dimethyl sulfoxide (DMSO; E70AGD). Five-millimeter bovine testicular tissue cubes could be completely penetrated by ethanol-based fixatives and Bouin solution in 2-3 h, whereas F4 required 21 h. Bouin solution produced general tissue shrinkage, whereas the other fixatives (alcohol-based and F4) caused tissue volume expansion. Although Bouin solution is an excellent fixative for testicular tissue, ethanol-based fixatives showed good penetration rates, low tissue shrinkage, and preserved sufficient morphology to allow identification of the stages of the seminiferous epithelium cycle, therefore representing a valid alternative for histotechnology laboratories. Common additives such as acetic acid, glycerol, and DMSO offered marginal benefits for the process of fixation; E70AG showed the best preservation of morphology with excellent nuclear detail, close to that of Bouin solution.
authors.affiliation: Ecuadorian Agency for Quality Assurance in Agriculture (AGROCALIDAD), Tumbaco, Ecuador (Cabrera, Espinoza, Vargas-Jentzsch, Sandoval, Ramos, Aponte).|Departamento de Ciencias Nucleares, Facultad de Ingeniería Química y Agroindustria, Escuela Politécnica Nacional, Quito, Ecuador (Vargas-Jentzsch).|Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito USFQ, Quito, Ecuador (Aponte).
authors.forename: Nelson C
authors.initials: NC
authors.lastname: Cabrera
authors.affiliation: Ecuadorian Agency for Quality Assurance in Agriculture (AGROCALIDAD), Tumbaco, Ecuador (Cabrera, Espinoza, Vargas-Jentzsch, Sandoval, Ramos, Aponte).|Departamento de Ciencias Nucleares, Facultad de Ingeniería Química y Agroindustria, Escuela Politécnica Nacional, Quito, Ecuador (Vargas-Jentzsch).|Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito USFQ, Quito, Ecuador (Aponte).
authors.forename: Jorge R
authors.initials: JR
authors.lastname: Espinoza
authors.affiliation: Ecuadorian Agency for Quality Assurance in Agriculture (AGROCALIDAD), Tumbaco, Ecuador (Cabrera, Espinoza, Vargas-Jentzsch, Sandoval, Ramos, Aponte).|Departamento de Ciencias Nucleares, Facultad de Ingeniería Química y Agroindustria, Escuela Politécnica Nacional, Quito, Ecuador (Vargas-Jentzsch).|Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito USFQ, Quito, Ecuador (Aponte).
authors.forename: Paul
authors.initials: P
authors.lastname: Vargas-Jentzsch
authors.affiliation: Ecuadorian Agency for Quality Assurance in Agriculture (AGROCALIDAD), Tumbaco, Ecuador (Cabrera, Espinoza, Vargas-Jentzsch, Sandoval, Ramos, Aponte).|Departamento de Ciencias Nucleares, Facultad de Ingeniería Química y Agroindustria, Escuela Politécnica Nacional, Quito, Ecuador (Vargas-Jentzsch).|Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito USFQ, Quito, Ecuador (Aponte).
authors.forename: Patricio
authors.initials: P
authors.lastname: Sandoval
authors.affiliation: Ecuadorian Agency for Quality Assurance in Agriculture (AGROCALIDAD), Tumbaco, Ecuador (Cabrera, Espinoza, Vargas-Jentzsch, Sandoval, Ramos, Aponte).|Departamento de Ciencias Nucleares, Facultad de Ingeniería Química y Agroindustria, Escuela Politécnica Nacional, Quito, Ecuador (Vargas-Jentzsch).|Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito USFQ, Quito, Ecuador (Aponte).
authors.forename: Luis A
authors.initials: LA
authors.lastname: Ramos
authors.affiliation: Ecuadorian Agency for Quality Assurance in Agriculture (AGROCALIDAD), Tumbaco, Ecuador (Cabrera, Espinoza, Vargas-Jentzsch, Sandoval, Ramos, Aponte).|Departamento de Ciencias Nucleares, Facultad de Ingeniería Química y Agroindustria, Escuela Politécnica Nacional, Quito, Ecuador (Vargas-Jentzsch).|Colegio de Ciencias Biológicas y Ambientales, Universidad San Francisco de Quito USFQ, Quito, Ecuador (Aponte).
authors.forename: Pedro M
authors.initials: PM
authors.lastname: Aponte
mesh_terms: D019342:Acetic Acid; D000818:Animals; D002417:Cattle; D000431:Ethanol; D005404:Fixatives; D005557:Formaldehyde; D008297:Male; D010851:Picrates; D013737:Testis / Q000473:pathology*; D016707:Tissue Fixation / Q000662:veterinary*
chemical_list: D005404:Fixatives; D010851:Picrates; D005557:Formaldehyde; C026239:Bouin's solution; D000431:Ethanol; D019342:Acetic Acid
keywords: Bovine; Tollens reagent; ethanol; fixatives; formaldehyde; testis