For calcium imaging experiments, cells were collected and incubated with 3 μM Fluo-3 AM for 15 min at 37 °C in the darkroom. Fluorescent images of Fluo-3AM-loaded cells were acquired using Cell Observer-Living Cells (Zeiss, German).
For calcium flux experiments, Fluo-3AM-loaded cells were centrifuged at 1000 rpm for 5 min at RT, washed once with PBS, and resuspended with 500 μl PBS. The intracellular calcium concentration was detected by flow cytometry (BD Biosciences, San Jose, CA, USA). The excitation source for Fluo-3 AM was a 488-nm air-cooled argon laser, and the emission was measured using a 525-nm band-pass filter.
For calcium flux experiments, Fluo-3AM-loaded cells were centrifuged at 1000 rpm for 5 min at RT, washed once with PBS, and resuspended with 500 μl PBS. The intracellular calcium concentration was detected by flow cytometry (BD Biosciences, San Jose, CA, USA). The excitation source for Fluo-3 AM was a 488-nm air-cooled argon laser, and the emission was measured using a 525-nm band-pass filter.
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