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TSQ fluorochrome incubation increase autometallographic zinc detection in primary cell cultures of astrocytes

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TSQ fluorochrome incubation increase autometallographic zinc detection in primary cell cultures of astrocytes

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dc.contributor.author Ballestín Hinojosa, Raúl
dc.contributor.author Torres, Jose Manuel
dc.contributor.author Ponsoda i Martí, Xavier
dc.date.accessioned 2022-04-08T07:07:25Z
dc.date.available 2022-04-09T04:45:06Z
dc.date.issued 2021 es_ES
dc.identifier.citation Póster 2021-10-26 XIX Congreso de la Sociedad Española de Biología Celular Madrid es_ES
dc.identifier.uri https://hdl.handle.net/10550/82229
dc.description.abstract In this work, we have analyzed the combination of direct (TSQ) and indirect (AMG) zinc detection methods in order to increase the sensitivity to a better study of the zinc uptake. For that purpose, rat primary cell cultured astrocytes were used. Intracellular Zn levels were visualized by using the TSQ zinc fluorochrome, either in normal conditions or after supplementary addition of 50 µM ZnSO4 to the culture. Fluorescence was recorded with an Olympus microscope BX50WI, equipped with a Hamamatsu ORCA digital camera controlled with the Aquacosmos software. Monolayers were fixed and ionic zinc precipitated by sodium sulphide or sodium selenite. Autometallographic zinc detection was the last step to detect zinc in cell cultures. Zincosome formation were visualized meanwhile the zinc fluorochrome TSQ in light microscopy, with or without adding zinc to cell monolayers. Additionally, we tested the better zinc precipitation and fixative method in order to preserve zinc ions and make them suitable for AMG development, selecting 0.1% sodium sulphide and 4% PFA. TSQ incubation demonstrated that are crucial for AMG zinc detection in both, prior and after the cell fixation. We found the setting parameters by which the zinc ion content of zincosomes can be maintained and visualized in optimal conditions and circumstances, either for optical and electron microscopy zinc detection studies. es_ES
dc.language.iso en es_ES
dc.subject astrocito es_ES
dc.subject cultivo celular es_ES
dc.subject zinc es_ES
dc.subject tsq es_ES
dc.subject autometalografía es_ES
dc.title TSQ fluorochrome incubation increase autometallographic zinc detection in primary cell cultures of astrocytes es_ES
dc.type conference output es_ES
dc.subject.unesco UNESCO::CIENCIAS DE LA VIDA es_ES
dc.accrualmethod - es_ES
dc.embargo.terms 0 days es_ES

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