Abstract
Confocal microscopy, coupled to high-performance hardware and software systems, has provided scientists with the capability of overcoming some of the limitations of standard microscopic imaging measurements of intracellular ions. The technique for loading of ion fluorescent probes is easily achieved; however, the quality of calcium measurements depends on the way of using the confocal system. In order to optimize this technique, scientists need to be familiar with the basic approaches and limitations of confocal microscopy. In this chapter, we will describe sample preparation, fluorescent probe loading, labeling of intracellular compartments, and the setting of parameters as well as protocols for measurements and limitations of the technique.
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Acknowledgments
This study was supported by grants from the Canadian Institutes of Health Research (CIHR), the Natural Sciences and Engineering Research Council of Canada (NSERC), and the Heart and Stroke Foundation of Quebec (HSFQ) (grants awarded to Drs. Ghassan Bkaily and Danielle Jacques).
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Bkaily, G., Al-Khoury, J., Simon, Y., Jacques, D. (2017). Intracellular Free Calcium Measurement Using Confocal Imaging. In: Touyz, R., Schiffrin, E. (eds) Hypertension. Methods in Molecular Biology, vol 1527. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6625-7_14
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DOI: https://doi.org/10.1007/978-1-4939-6625-7_14
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