Phagocytosis of FAM-labeled Amyloid-β

Phagocytosis was examined via FSX100 Bio Imaging Navigator (Olympus Waltham, MA) using a Human β-Amyloid (1–42), 5-FAM-labeled according to the manufacturer’s protocol. Primary microglial cells cultured in glass-based dishes were used. Human β-Amyloid (1–42), 5-FAM-labeled was reconstituted and diluted with 1X PBS and to a concentration of 3 µg/mL according to both the manufacturer’s protocol and previous reports^{40 (link),41 (link)}. We incubated the cells in standard culture conditions for 3 h. After aspirating the culture medium, the cells were fixed with 4% paraformaldehyde. Then, after discarding paraformaldehyde, we washed fixed cells with 1 mL phosphate buffered salts (PBS) twice and measured the fluorescence intensity of FAM along the long axis of the cytoplasm using an Imaging Navigator.