Dear List members Greetings I would like to ask you some questions about the use of EDTA as demineratizing agent. I have to do histology and histochemistry (including metallothioneins) in fish larvae preserved in Bouins solution.
1) First of all I would like to know what is the best formula (or the one you recomend) to prepare the EDTA solution, as well as the importance of maintaining a given pH and if it is necessary to put the samples in the refrigerator during the demineralization process. 2) Is EDTA recomended only for certain fixatives, or It can be used with any fixative, including Bouins 3) When using Bouins solution as fixative, many books recommend not to wash in water but instaed to remove the excess of picric acid using several washes in etanol 70%, on the other hand, I´ve read that EDTA precipitates in the presence of alcohols. What do you recomend in this case? 4) Do you think that the demineralization procedure could alter or affect the detection of metallothioneins in the tissues? Many thanks in advance Best regards José Luis Dr. José Luis Palazón Fernández Instituto de Investigaciones Científicas Universidad de Oriente Boca del Rio-Isla Margarita-Venezuela Direccion actual: Instituto de Ciencias Marinas de Andalucia-CSIC Campus universitario Rio San Pedro, 11510, Puerto Real, Cádiz, España email: jluis.pala...@icman.csic.es; jose.pala...@ne.udo.edu.ve tlf: +34-956832612; fax: +34-956834701; cell: +34-600487100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
