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Inmunomarcación de neuronas dopaminérgicas en cortes flotantes de hipotálamo de rata: preservación alternativa del tejido nervioso antes del corte

Keywords: immunohistochemistry, tyrosine hydroxylase, free floating sections, cryopreservation.

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Abstract:

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.

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