Intracellular sodium concentration ([Na+](i)) can be measured using SB
FI with image analysis or spectrofluorometry. However, SBFI cannot be
used with most now cytometry systems, due to its requirement for dual
excitation in the UV range. Recently a new sodium indicator, Sodium Gr
een, was developed which can be used with now cytometry to measure [Na
+](i). Adequate staining of cells with Sodium Green is dependent upon
both dye and cell concentration, time and temperature of loading, use
of the non-ionic detergent Pluronic, and the cell Line used. Both Sodi
um Green and SBFI give similar calibration curves using the ionophore
gramicidin, Sodium Green has a small pH dependence, but this should no
t affect most applications, Since a fluorescence ratio technique canno
t be used with Sodium Green, we used a ratio of dye fluorescence to fo
rward angle Light scatter to normalize for cell size variations in the
population, The basal [Na+](i) obtained using the Sodium Green techni
que in CHO IS1 cells was 19 mM, in reasonable agreement with previous
studies using SBFI in other cells. (C) 1995 Wiley-Liss, Inc.