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.

Na+–H+exchanger and proton channel in heart failure associated with Becker and Duchenne muscular dystrophies

“…It is worth noting that all pharmacological therapies done in hereditary cardiomyopathy and more specifically using the hereditary cardiomyopathic hamster model were to prevent necrosis that takes place during the early development of HF, and no studies were done to verify the effect of all the drugs used on HF leading to early death. Nevertheless, all of these early studies raised the issue that calcium channels do not necessarily contribute to the development of HF in the animal model of DMD and BMD (Bkaily et al 1996(Bkaily et al , 2017. This question was partially answered by the discovery of the early fetal TTX-and Mn ++ -insensitive slow sodium channel in both fetal human and chick embryo heart cells (Bkaily et al 1990(Bkaily et al , 2017.…”

“…Nevertheless, all of these early studies raised the issue that calcium channels do not necessarily contribute to the development of HF in the animal model of DMD and BMD (Bkaily et al 1996(Bkaily et al , 2017. This question was partially answered by the discovery of the early fetal TTX-and Mn ++ -insensitive slow sodium channel in both fetal human and chick embryo heart cells (Bkaily et al 1990(Bkaily et al , 2017. This channel was found to have, to some extent, a pharmacology similar to that of the L-type calcium channel because it was blocked by verapamil (Bkaily et al 1990(Bkaily et al , 2017, which was shown to prevent necrosis and hypertrophy in UM-X7.1 hamsters (Jasmin et al 1991).…”

“…1), intracellular sodium overload (Fig. 2) is one of the earliest pathological signs (Bkaily et al , 2017Chahine et al 2005;Jacques et al 2003). The latter was found to be, in part, a consequence of the development of intracellular acidosis and the resulting sodium influx via the Na + -H + exchanger NHE-1 (Fig.…”

“…The latter was found to be, in part, a consequence of the development of intracellular acidosis and the resulting sodium influx via the Na + -H + exchanger NHE-1 (Fig. 2) (Bkaily et al 2015(Bkaily et al , 2017. Treatment with an NHE-1 blocker, EMD87580, not only prevented intracellular sodium overload, but also prevented the increase of circulating creatine kinase and early death due to HF (Bkaily et al 2015).…”

“…2) awaits elucidation. For further information concerning sodium overload and NHE-1 in DMD and BMD, the reader should refer to key references (Bkaily et al 1990(Bkaily et al , 2015(Bkaily et al , 2017Al-Khoury et al 2006;Chahine et al 2005;Jacques et al 1997Jacques et al , 2003Honoré et al 2008). …”