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The in situ K was 1.652×10 μM (n=58 fibres, R =0.99). With an F of 150.9 (8.8) A.U. (n=8), F of 0.14 (0.1) A.U. (n=1, and ΔF of Ca transients of 8.4 (2.5) A.U. (n=1, the sarcoplasmic [Ca ] reached 22.5 (7.8) μM. Compartmentalized dye amounted to only 1.1 (0.7)% (n=1. Two Mag-Fluo-4 molecules coalesce around one Ca ion, in an entropy-driven, very low in situ affinity reaction, making it suitable to reliably track the kinetics of rapid muscle Ca transients. Our results may be relevant to the quantitative study of Ca kinetics in many