Cell experiment:

For measuring fluorescence from cells in suspension, dilutions to 2 to 3×106 cells are made, from cultures of rat basophilic leukemia (RBL) cells. Cells are incubated in suspension in 1 mM dye (including Fluo-4 AM) for 30 min at 37℃. Cell suspensions are then transferred to cuvets for measurements of fluorescence emission intensity by spectrofluorometer[1].

Fluo-4 AM is a fluorescent dye (λex=494 nm, λem=516 nm).

Fluo-4 AM is a fluorescent dye (λex=494 nm, λem=516 nm). Preloaded with Fuo-4 AM, a very bright fluorescence image is observed. In a parallel experiment with fluo-3 AM-loaded cells, the resulting fluorescence image, although clearly discernable in this case, is less bright[1].

[1]. Gee KR, et al. Chemical and physiological characterization of fluo-4 Ca(2+)-indicator dyes. Cell Calcium. 2000 Feb;27(2):97-106.