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使用比率荧光染料SNARF-5F-AM对涡虫进行细胞内pH的实时成像。

Live imaging of intracellular pH in planarians using the ratiometric fluorescent dye SNARF-5F-AM.

作者信息

Beane Wendy Scott, Adams Dany Spencer, Morokuma Junji, Levin Michael

机构信息

Department of Biology and Center for Regenerative and Developmental Biology, Tufts University, Medford, MA 02155, USA.

出版信息

Biol Methods Protoc. 2019;4(1):bpz005. doi: 10.1093/biomethods/bpz005. Epub 2019 May 29.

Abstract

Physiological parameters such as resting potential and pH are increasingly recognized as important regulators of cell activity and tissue-level events in regeneration, development, and cancer. The availability of fluorescent reporter dyes has greatly increased the ability to track these properties . The planarian flatworm is an important and highly tractable model system for regeneration, stem cell biology, and neuroscience; however, no protocols have been published for investigating pH in this system. Here, we report a simple and effective protocol for imaging pH gradients in living planaria suitable for intact and regenerating flatworms.

摘要

静息电位和pH值等生理参数越来越被认为是再生、发育和癌症中细胞活动和组织水平事件的重要调节因子。荧光报告染料的可用性大大提高了追踪这些特性的能力。涡虫是用于再生、干细胞生物学和神经科学研究的重要且易于处理的模型系统;然而,尚未有针对该系统中pH值研究的方案发表。在此,我们报告了一种简单有效的方案,用于对适合完整和再生涡虫的活涡虫体内pH梯度进行成像。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/83df/7045346/bdd500444a48/bpz005f1.jpg

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