Rijkers G T, Justement L B, Griffioen A W, Cambier J C
Department of Immunology, University Hospital for Children and Youth, Het Wilhelmina Kinderziekenhuis, Utrecht, The Netherlands.
Cytometry. 1990;11(8):923-7. doi: 10.1002/cyto.990110813.
The accuracy of flow cytometric measurement of intracellular calcium with fluo-3 is compromised by variation in basal fluorescence intensity due to heterogeneity in dye uptake or compartmentalization. We have loaded cells simultaneously with fluo-3 and SNARF-1. When SNARF-1 fluorescence is collected at approximately 600 nm, its intensity does not change upon cell activation. Furthermore, fluo-3 and SNARF-1 fluorescence signals exhibit a linear relationship. The ratio of fluo-3 to SNARF-1 eliminates a significant proportion of variation in fluorescence intensity caused by variation in fluo-3 uptake and thus can be used as a sensitive parameter for measuring changes in [Ca2+]i.
由于染料摄取或区室化的异质性导致基础荧光强度变化，从而影响了用Fluo-3进行细胞内钙的流式细胞术测量的准确性。我们同时用Fluo-3和SNARF-1加载细胞。当在约600nm处收集SNARF-1荧光时，其强度在细胞激活时不会改变。此外，Fluo-3和SNARF-1荧光信号呈现线性关系。Fluo-3与SNARF-1的比率消除了因Fluo-3摄取变化引起的荧光强度变化的很大一部分，因此可以用作测量[Ca2+]i变化的敏感参数。
