Monitoring Amyloid-Beta Induced Calcium Dynamics

2 × 10⁵ HT22 cells were incubated in 35 mm confocal dishes for 24 h at 37 °C in a CO² incubator. A 5 mM Fluo-3 AM stock solution was diluted with Hanks-Balanced Salt Solution (HBSS) to a 5 μM working solution and then added to the cells for 30 min at 37 °C. HT22 cells were washed three times with HEPES-buffered saline and then incubated in an imaging chamber at 37 °C for an additional 10 min. After the addition of 30 μM of Aβ aggregates in HBSS buffer, changes in cellular Ca²⁺ levels were monitored by epifluorescence microscopy (Applied Precision DeltaVision Elite, Applied Precision, Inc., United States) using the real-time mode for tracking Fluo-3 changes for 5 min. The data inspection program provided by DeltaVision software was used to measure the intensity of Fluo-3 fluorescence.