Ratiometric Imaging of pH Probes

• Published in: Methods in Cell Biology Vol. 123; pp. 429 - 448
• Main Authors: Grillo-Hill, Bree K., Webb, Bradley A., Barber, Diane L.
• Format: Book Chapter Journal Article
• Language: English
• Published: United States Elsevier Science & Technology 2014
• Subjects: Animals; Biosensing Techniques - methods; Biosensing Techniques - standards; Cell Line; Cellular biology; Drosophila melanogaster; Fluoresceins - chemistry; Fluorescent Dyes - chemistry; Genetically encoded pH biosensor; Green Fluorescent Proteins - metabolism; Hydrogen-Ion Concentration; In situ imaging; Live-cell imaging; Microscopy, Fluorescence; pH measurement; pH-sensitive dyes; Quantitative imaging; Ratiometric imaging; Reference Standards; Single-Cell Analysis; Whole-mount tissue imaging; Ratiometric imaging; pH-sensitive dyes; Genetically encoded pH biosensor; In situ imaging; Quantitative imaging; Live-cell imaging; pH measurement; Drosophila melanogaster; Whole-mount tissue imaging
• ISBN: 9780124201385; 0124201385
• ISSN: 0091-679X
• DOI: 10.1016/B978-0-12-420138-5.00023-9

Measurement of intracellular pH can be readily accomplished using tools and methods described in this chapter. We present a discussion of technical considerations of various ratiometric pH-sensitive probes including dyes and genetically encoded sensors. These probes can be used to measure pH across physical scales from macroscopic whole-mount tissues down to organelles and subcellular domains. We describe protocols for loading pH-sensitive probes into single cells or tissues and discuss ratiometric image acquisition and analysis.