2.10. Phagocytic Activity Determination

The phagocytic activity of alveolar macrophages was determined by the Vybrant™ Phagocytosis Assay Kit (Thermo Fisher). According to the manufacturer's instruction, BALF cells were collected from the lungs and cultured in the 96-plat bottom-well plates in the presence of fluorescein-labelled Escherichia coli K-12 BioParticles. One hour later, the BioParticle loading suspension from all of the microplate wells was removed by vacuum aspiration. The extracellular fluorescence probe was then quenched off by trypan blue. The microplate was read by the fluorescence plate reader (Spark® Multimode Microplate Reader, Tecan).