| Chapter title |
Measuring Phagosomal pH by Fluorescence Microscopy.
|
|---|---|
| Chapter number | 12 |
| Book title |
Phagocytosis and Phagosomes
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6581-6_12 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6579-3, 978-1-4939-6581-6
|
| Authors |
Johnathan Canton, Sergio Grinstein |
| Editors |
Roberto Botelho |
| Abstract |
Dual wavelength ratiometric imaging has become a powerful tool for the study of pH in intracellular compartments. It allows for the dynamic imaging of live cells while accounting for changes in the focal plane, differential loading of the fluorescent probe, and photobleaching caused by repeated image acquisitions. Ratiometric microscopic imaging has the added advantage over whole population methods of being able to resolve individual cells and even individual organelles. In this chapter we provide a detailed discussion of the basic principles of ratiometric imaging and its application to the measurement of phagosomal pH, including probe selection, the necessary instrumentation, and calibration methods. |
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