摘要
应用Fura-2测定急性白血病(急白)骨髓或外周血中单个核细胞内钙离子浓度([Ca^(2+)]i)。肝素化血或骨髓液与RPMI1640混合后,滴入淋巴细胞分层液,离心。分离出的单个核细胞与Fura-2 AM,BSA,37℃温育40min。按激发波长350nm,发射波长500nm,测量其荧光强度F,Fmax,Fmin。用本法测定35例急白患者(51例次),结果表明[Ca^(2+)]i变化可作为一项反映化疗药物敏感性、预测化疗疗效的参考指标。
This paper describes a fluorimetric method with Fura-2 as indicater to determine the intracellular ionic calcium concentration of mononuclear cells from the bone marrow or the blood of patients with acute leukemia. The heparinized blood or bono marrow was mixed with medium RPMI-1640. then dropped on a Ficoll--metrizonate mixture. The mononuclear cells separated by centrifugation were incubated of 37℃ for 40min with Fura-2 AM and BSA. Fluorescorce was measured with a spectrofluo- rimeter. Excitation and emission wavelengths were 350 nm and 500 nm, respectively. The free calcium concentration was calculated by the equation [Ca^(2+)]i=Kd (F--Fmin )/ (Fmax--F). Using this method in 53 clinical patients, a satisfactory result has been achieved.
关键词
白血病
核细胞
钙离子
acute leukemia
Fura--2
intracellular free calcium
fluorimetric method
