Phagocytosis of pHrodo E.coli BioParticles by microglial cells

We wanted to study phagocytosis by microglial cells. In our study we wanted to specifically treat cells with different stress inducers and observe the effect of such stress on microglial phagocytotic ability. For this experiment we used BV-2 and N9 mouse microglial cell line and plated them in poly-L lysine coated glass cover slips in 24 well plates. In the meantime, the pHrodo particles were mixed in 1X PBS (pH 7.4) and sonicated for 5 minutes at high frequency. 24 hours later the pHrodo particles were nicely vortexed and 20 µl of solution was added to each well. Phagocytosis was allowed for 2 hours and then cells were fixed and co-stained with hoechst dye.