[1]程颖,ABDULLA S.,CHORVAT D. J. R.,等.基于多维TCSPC技术的心肌细胞Ca2+动力学研究[J].深圳大学学报理工版,2008,25(4):369-375.
 CHENG Ying,ABDULLA S.,CHORVAT D. J. R.,et al.Investigation of intracellular calcium dynamics during cardiac cell contraction by multi-dimensional TCSPC[J].Journal of Shenzhen University Science and Engineering,2008,25(4):369-375.
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基于多维TCSPC技术的心肌细胞Ca2+动力学研究()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第25卷
期数:
2008年4期
页码:
369-375
栏目:
生物医学工程
出版日期:
2008-10-31

文章信息/Info

Title:
Investigation of intracellular calcium dynamics during cardiac cell contraction by multi-dimensional TCSPC
文章编号:
1000-2618(2008)04-0369-07
作者:
程颖14ABDULLA S.1CHORVAT D. J. R.3CHORVATOVA A.12
1)蒙特利尔大学医学中心圣-贾斯廷医院研究中心,蒙特利尔H3T 1C5,加拿大
2)蒙特利尔大学儿科部,蒙特利尔H3T 1C5,加拿大
3)国际激光中心,伯拉第斯拉瓦81219,斯洛伐克
4)北京大学深圳医院心血管外科,深圳 518036,中国
Author(s):
CHENG Ying14ABDULLA S.1CHORVAT D. J. R.3and CHORVATOVA A.12
1)Research Centre of CHU Sainte-Justine,University of Montreal,Montreal H3T 1C5,Canada
2)Department of Pediatrics,University of Montreal,Montreal H3T 1C5,Canada
3)International Laser Centre,Bratislava 81219,Slovakia
4)Department of Cardiovascular Surgery,Beijing University Shenzhen Hospital,Shenzhen 518036,P.R.China
关键词:
时间相关单光子计数线粒体Ca2+标记荧光心肌收缩
Keywords:
time correlated single photon counting (TCSPC)mitochondriaCalcium-sensitive fluorescence probescardiomyocyte contraction
分类号:
TP 271.+5
文献标志码:
A
摘要:
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540 nm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.
Abstract:
Spectrally-resolved time correlated single photon counting (TCSPC) technique was applied to investigate the changes of mitochondrial vs cytosolic calcium. Calcium-sensitive fluorescence probes Fluo-4 and Ca-orange were used to test cytosolic and/or mitochondrial calcium dynamics. The results show that upon comparison with steady state conditions (measured 1 s after the beginning of contraction), Fluo-4 fluorescence intensity is significantly higher when measured at 10~110 ms after the start of contraction, as calcium is elevated in contracting myocytes. After normalization, time-resolved spectra of the Fluo-4 were measured and corrected by cell autofluorescence. The spectral maximum was at 540 nm. For cells in steady state conditions labeled by Fluo-4, the fluorescence lifetimes and their relative amplitudes at emission wavelength of 540 nm were identified:τ1= 0.42±0.01 ns (73±5)% and τ2 =2.74±0.67 ns (25±5)% yielding the mean lifetime of the decay τmean= 0.83 ns. The emission maximum of mitochondrial probe Ca-orange was at 560 nm. At the same time, Rotenone, inhibitor of Complex I of the mitochondrial respiratory chain, was used and significantly decreased the fluorescence intensity at 500~580 nm range, measured at 10~110 ms after the start of contraction. We evaluated spectrally-and time-resolved fluorescence characteristics of cytosolic and mitochondrial calcium-sensitive probes Fluo-4 and Ca-orange, respectively, in contracting cardiac myocytes. This new approach provides satisfactory spectrally-and time-resolved fluorescence recordings during cardiomyocyte contraction cycles.

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备注/Memo

备注/Memo:
收稿日期:2008-05-29;修回日期:2008-09-18
基金项目:加拿大魁北克省心脏和中风基金资助项目(APVV-51-037905);加拿大创新基金资助项目(9684)
作者简介:程颖(1964-),男(汉族),安徽省黄山市人, 北京大学深圳医院心血管外科副主任医师. E-mail:chengying627@hotmail.com
*本文为2008第3届国际生物医学光学方法高级研讨会宣读论文
更新日期/Last Update: 2008-11-26