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

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 st…