title: Intracellular Free Calcium Measurement Using Confocal Imaging.
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.
authors.affiliation: Department of Anatomy and Cell Biology, Faculty of Medicine, University of Sherbrooke, 2500 boul. De l'Universite, Sherbrooke, QC, Canada, JIH 5N4. Ghassan.Bkaily@usherbrooke.ca.
authors.forename: Ghassan
authors.initials: G
authors.lastname: Bkaily
authors.affiliation: Department of Anatomy and Cell Biology, Faculty of Medicine, University of Sherbrooke, 2500 boul. De l'Universite, Sherbrooke, QC, Canada, JIH 5N4.
authors.forename: Johny
authors.initials: J
authors.lastname: Al-Khoury
authors.affiliation: Department of Anatomy and Cell Biology, Faculty of Medicine, University of Sherbrooke, 2500 boul. De l'Universite, Sherbrooke, QC, Canada, JIH 5N4.
authors.forename: Yanick
authors.initials: Y
authors.lastname: Simon
authors.affiliation: Department of Anatomy and Cell Biology, Faculty of Medicine, University of Sherbrooke, 2500 boul. De l'Universite, Sherbrooke, QC, Canada, JIH 5N4.
authors.forename: Danielle
authors.initials: D
authors.lastname: Jacques
mesh_terms: D000814:Aniline Compounds / Q000032:analysis; D002118:Calcium / Q000032:analysis*; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D007211:Indoles / Q000032:analysis; D018613:Microscopy, Confocal / Q000379:methods*; D014966:Xanthenes / Q000032:analysis
chemical_list: D000814:Aniline Compounds; C409648:Fluo 4; D007211:Indoles; D014966:Xanthenes; C059715:Fluo-3; C048960:indo-1; D002118:Calcium
keywords: Calcium probe; Confocal microscopy; Fluo-3; Fluo-4; Indo-1; Intracellular calcium; Nuclear calcium