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Cellular localization of GO and GNPs by confocal microscopy

JD Josefa Domenech
AH Alba Hernández
ED Esref Demir
RM Ricard Marcos
CC Constanza Cortés
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After the exposure to GO/GNPs, the monolayers’ nuclei and mucus were stained at room temperature with Hoechst 33342 (ThermoFisher Scientific, USA) at a 1:500 dilution and with Wheat Germ Agglutinin Alexa Fluor 633 (WGA) Conjugate (ThermoFisher Scientific, USA) diluted 1:100, respectively, for 15 min. Afterward, the barriers were washed with DMEM twice, the Transwell’s membrane was cut with a scalpel and placed in Glass Bottom Microwell Dishes (MatTek, USA). GO/GNPs were identified thanks to their light-reflection properties. The images of samples (n = 2) were taken with a Leica TCS SP5 confocal microscope and processed with Imaris 8.2.1 software (Bitplane, AG) to manually mask mucus, nanoparticles, and nuclei in red, green, and blue, respectively.

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