Next, 25 µl of fluorescein-5-isothyocianate FITC diluted to 1/10 with PBS and 2 µl of Evans Blue 0.5% were placed on each coverslip on the microplates, followed by incubation in a humid atmosphere for 30 min at 37°C. Three washings were then performed using PBS for 5 min each, and the coverslips were removed from each well and the contents were allowed to dry at room temperature Vanrompay et al., 1994.
Se tomaron 25 µl de los anticuerpos marcados con fluoresceína-5-isotiocianato, se diluyeron 1/10 en PBS y se agregaron 2 µl de azul de Evans 0.5% para contrastar la tinción; éstos se añadieron a cada pozo de la placa, seguido de la incubación en una cámara húmeda durante 30 minutos a 37 °C. Se lavaron tres veces durante cinco minutos con PBS y los cubreobjetos se sacaron de los pocillos y se dejaron secar a temperatura ambiente Vanrompay et al., 1994.