在成年大鼠心肌细胞中校准和测量线粒体 pH 值。

Calibration and measurement of mitochondrial pH in intact adult rat cardiomyocytes.

机构信息

Institute for Regenerative Medicine, Shanghai East Hospital, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.

Department of Pharmacy, Shanghai East Hospital, Tongji University, Shanghai 200120, China.

出版信息

STAR Protoc. 2021 May 14;2(2):100543. doi: 10.1016/j.xpro.2021.100543. eCollection 2021 Jun 18.

DOI:10.1016/j.xpro.2021.100543

PMID:34036286

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8138862/

Abstract

Mitochondrial pH is a vital parameter of the mitochondrial environment, which determines the rate of many mitochondrial functions, including metabolism, membrane potential, fate, etc. Abnormal mitochondrial pH is always closely related to the health status of cells. Analyzing mitochondrial pH can serve as a proxy for mitochondrial and cellular function. This protocol describes the use of SNARF-1 AM, a pH-sensitive fluorophore, to measure mitochondrial pH. This protocol details the steps to evaluate mitochondrial pH in live adult cardiomyocytes using confocal microscopy. The protocol can be adapted to other adherent cell types. For complete details on the use and execution of this protocol, please refer to Wei-LaPierre et al. (2013).

摘要

线粒体 pH 是线粒体环境的一个重要参数，它决定了许多线粒体功能的速率，包括代谢、膜电位、命运等。异常的线粒体 pH 总是与细胞的健康状况密切相关。分析线粒体 pH 可以作为线粒体和细胞功能的替代指标。本方案描述了使用 SNARF-1 AM，一种 pH 敏感的荧光染料，来测量线粒体 pH。本方案详细介绍了使用共聚焦显微镜评估活成年心肌细胞中线粒体 pH 的步骤。该方案可以适应其他贴壁细胞类型。有关使用和执行本方案的完整详细信息，请参见 Wei-LaPierre 等人（2013 年）。