Acta toxicológica argentina
versión On-line ISSN 1851-3743
CHOLICH, Valeria; GARCIA, Graciela; MARTINEZ, Alejandra y EVANGELISTA DE DUFFARD, Ana María. Immunohistochemical Studies Of Dopaminergic Neurons On Free Floating Sections: Alternative Cryopreservation Method Of Nervous Tissue Before Cutting. Acta toxicol. argent. [online]. 2008, vol.16, n.1, pp. 1-4. ISSN 1851-3743.
In central nervous system histological studies, free-floating sections of perfusion-fixed samples are frequently used. Samples storage may be performed freezing either the entire brain at -80ºC or sections at -20ºC. When studying hypothalamic tyrosine hydroxylase enzyme (dopaminergic neurons marker) by immunohistochemistry in adult male Wistar rats, entire brains were stored at -80ºC. Due to an abrupt freezer technical failure, samples should be thawed to -20ºC with the resulting samples damage. To avoid this situation, subsequent brains were stored in 30% sucrose in saline phosphate buffer (PBS) with 0.01% sodium azide and kept at 4ºC for different periods (weeks to months) until they were frozen with chlorofluorade gas and cut. These brains showed no morphological alterations of tissue structure. This preservation method appeared to be an alternative valid option to laboratories with no -80ºC freezing equipment.
Palabras llave : Immunohistochemistry; Tyrosine hydroxylase; Free floating sections; Cryopreservation.