过氧化物酶体
细胞器
共焦显微镜
细胞生物学
荧光显微镜
细胞内
共焦
显微镜
活体细胞成像
生物
荧光
生物物理学
细胞
化学
生物化学
光学
基因
物理
作者
Luís F. Godinho,Michael Schrader
出处
期刊:Springer eBooks
[Springer Nature]
日期:2017-01-01
卷期号:: 181-189
被引量:5
标识
DOI:10.1007/978-1-4939-6937-1_16
摘要
Organelle pH homeostasis is crucial for maintaining proper cellular function. The nature of the peroxisomal pH remains somewhat controversial, with several studies reporting conflicting results. Here, we describe in detail a rapid and accurate method for the measurement of peroxisomal pH, using the pHRed sensor protein and confocal microscopy of living mammalian cells. pHRed, a ratiometric sensor of pH, is targeted to the peroxisomes by virtue of a C-terminal targeting sequence. The probe has a maximum fluorescence emission at 610 nm while exhibiting dual excitation peaks at 440 and 585 nm, allowing for ratiometric imaging and determination of intracellular pH in live cell microscopy.
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