摘要
用线粒体膜电位特异性荧光探针Rhodamine123标记线粒体后 ,在改装FACS420流式细胞仪上用C -30数据采集和储存程序采集并存储数据 ,用C -30分析程序分析得到反映线粒体前散射 (FSC)、Rhodamine123荧光强度和侧散射 (SSC)变化的FSC,FL1和FL2合并图。结果显示 ,200μmol/LCa2 + 处理20分钟使线粒体同时发生FSC增大、Rhodamine123荧光强度降低和SSC增大。结果表明 ,流式细胞术可同时检测到Ca2 +诱导PTP开放产生的线粒体膨胀 ,膜电位消失 ,线粒体颗粒性状改变。本方法可多指标测定 ,实验过程快速 ,结果准确可靠 ,所需样品量小 ,是一种理想的PTP开放检测手段。
Mitochondria potential sensitive fluorescence probe rhodamine 123 was used to label the mitochondria isolated from rat liver. The changes in FSC, rhodamine 123 fluorescence and SSC of the mitochondria, which represented the size, the membrane potential and the granularity of mitochondria respectively, were determined by a modified FACS420 cytometer. C-30 software was used for data acquisition, storage and analyses. The results showed, stimulated with 200μmol/L Ca2+ for 20 minutes, the mitochondria displayed a concomitant increase in FSC and SSC, and decrease in fluorescence intensity. The results suggested that PTP opening induced by Ca2+ can be detected by cytometry, which is characterized by the distinguished features, such as speedy, accuracy, multi-index measurement, and less sample consumption, etc.
出处
《生物物理学报》
CAS
CSCD
北大核心
2000年第2期225-230,共6页
Acta Biophysica Sinica
基金
中国科学院"九五"重大课题!(KJ951 -B1 -609)
中国博士后基金资助项目
