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。