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一种使用钙绿-2记录心肌细胞内[Ca2+]瞬变的方法。

A method for recording intracellular [Ca2+] transients in cardiac myocytes using calcium green-2.

作者信息

Spencer C I, Berlin J R

机构信息

Bockus Research Institute, Graduate Hospital, Philadelphia, PA 19146, USA.

出版信息

Pflugers Arch. 1995 Aug;430(4):579-83. doi: 10.1007/BF00373895.

DOI:10.1007/BF00373895
PMID:7491285
Abstract

Calcium green-2 (Ca green) is a non-ratiometric fluorescent Ca2+ indicator with an affinity for Ca2+ (dissociation constant Kd = 3 microM) that is lower than more commonly used indicators such as fura-2 and fluo-3. This low Ca2+ affinity, coupled with a high quantum yield, allows cells to be loaded with low concentrations of Ca green, avoiding problems of cytosolic Ca2+ buffering and a low signal-to-noise ratio. This communication presents a method for monitoring intracellular [Ca2+] changes in isolated rat ventricular myocytes loaded with Ca green and the fluorescent pH indicator carboxy SNARF-1 (SNARF). SNARF provides a Ca(2+)-insensitive signal with which Ca green fluorescence can be corrected for cell motion and dye-loading artifacts.

摘要

钙绿-2(Ca green)是一种非比率型荧光Ca2+指示剂,对Ca2+具有亲和力(解离常数Kd = 3 microM),低于常用的指示剂如fura-2和fluo-3。这种低Ca2+亲和力与高量子产率相结合,使得细胞能够以低浓度加载钙绿,避免了胞质Ca2+缓冲问题和低信噪比。本通讯介绍了一种监测负载钙绿和荧光pH指示剂羧基SNARF-1(SNARF)的离体大鼠心室肌细胞内[Ca2+]变化的方法。SNARF提供了一个对Ca(2+)不敏感的信号,利用该信号可对钙绿荧光进行校正,以消除细胞运动和染料加载伪影的影响。

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A method for recording intracellular [Ca2+] transients in cardiac myocytes using calcium green-2.一种使用钙绿-2记录心肌细胞内[Ca2+]瞬变的方法。
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Control of sarcoplasmic reticulum calcium release during calcium loading in isolated rat ventricular myocytes.离体大鼠心室肌细胞钙负荷期间肌浆网钙释放的调控
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本文引用的文献

1
Ratiometric confocal Ca(2+)-measurements with visible wavelength indicators in isolated cardiac myocytes.在分离的心肌细胞中使用可见波长指示剂进行比率共聚焦钙测量。
Cell Calcium. 1993 May;14(5):359-72. doi: 10.1016/0143-4160(93)90040-d.
2
Ca transients in cardiac myocytes measured with a low affinity fluorescent indicator, furaptra.用低亲和力荧光指示剂呋喃普拉测定心肌细胞中的钙瞬变。
Biophys J. 1993 Apr;64(4):1331-43. doi: 10.1016/S0006-3495(93)81494-2.
3
Fluorescent properties of rat cardiac trabeculae microinjected with fura-2 salt.
用fura-2盐显微注射大鼠心脏小梁的荧光特性。
Am J Physiol. 1993 Apr;264(4 Pt 2):H1098-110. doi: 10.1152/ajpheart.1993.264.4.H1098.
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Ca2+ transients in cardiac myocytes measured with high and low affinity Ca2+ indicators.使用高亲和力和低亲和力钙指示剂测量心肌细胞中的钙瞬变。
Biophys J. 1993 Oct;65(4):1632-47. doi: 10.1016/S0006-3495(93)81211-6.
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J Physiol. 1988 Sep;403:151-92. doi: 10.1113/jphysiol.1988.sp017244.
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Effect of membrane potential changes on the calcium transient in single rat cardiac muscle cells.
Science. 1987 Dec 4;238(4832):1419-23. doi: 10.1126/science.2446391.
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Intracellular Ca transients in rat cardiac myocytes: role of Na-Ca exchange in excitation-contraction coupling.
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