论文部分内容阅读
采用Fluo-4和Ca-orange标记细胞质及线粒体中的Ca2+,通过多维时间相关单光子计数技术研究心肌收缩时Ca2+动力学.结果表明,在心肌收缩后10～110 ms的Fluo-4荧光强度明显强于心肌收缩后1 s的荧光强度,归一化光谱和细胞自发荧光一致;静止状态下,成分荧光寿命时间和相关振幅在峰值540nm波长处τ1=(0.42±0.01)ns(73±5)%,τ2=(2.74±0.67)ns(25±5)%,平均荧光衰减时间τmean=0.83 ns;Ca-orange荧光峰值560 nm,加入线粒体呼吸链阻断剂Rotenone后,10～110 ms的Ca-orange荧光强度显著降低.综合评估了心肌细胞收缩时细胞质和线粒体中荧光团的光谱及时间分辨荧光特性,该方法可为心肌收缩过程提供满意的光谱和时间分辨荧光记录,有助于理解细胞综合行为.
The Ca2 + kinetics of myocardial contractions were investigated by multi-dimensional time-dependent single-photon counting using Fluo-4 and Ca-orange labeling of Ca2 + in the cytoplasm and mitochondria. The results showed that Fluo-4 fluorescence intensity ten ~ 110 ms after myocardial contraction was significant The fluorescence intensity at 1 s after myocardial contraction was stronger than that at 1 s, and the normalized fluorescence spectrum was consistent with the autofluorescence of cells. At rest, the fluorescence lifetime and relative amplitudes of the components at the peak wavelength of 540 nm were τ1 = (0.42 ± 0.01) ns (73 ± 5) %, τ2 = (2.74 ± 0.67) ns (25 ± 5)%, the average fluorescence decay time τmean = 0.83 ns; Ca-orange fluorescence peak 560 nm. After adding the mitochondrial respiratory chain blocker Rotenone, -orange Fluorescence intensity was significantly reduced Spectral and time-resolved fluorescence characteristics of the fluorophores in the cytoplasm and mitochondria during cardiomyocyte contraction were evaluated in a comprehensive manner that provided satisfactory spectroscopic and time-resolved fluorescence recordings of myocardial contractile processes and helped to understand cells Comprehensive behavior.