Abstract

Publisher Summary This chapter describes the use of Ca2+-sensitive fluorescent dye technology with the extremely precise technology of flow cytometry for the single-cell analysis of changes in intracellular Ca2+ levels. The model cell involves the antireceptor stimulation of mouse splenic B lymphocytes, although the techniques described are readily adaptable to any cell type, which can be analyzed by flow cytometry and is not incompatible with the quin 2 or indo-1. The chapter also presents the relevant principles of flow cytometry. It is important to be familiar with the fundamental principles of flow cytometry to understand the techniques to be described and to permit their adaptation to the various instruments currently in use. The basic principle of the analytic system is to pass a monodisperse cell suspension (loaded with fluorescent dye) through a quartz flow cell, which is intersected by a focused laser beam of appropriate excitation wavelength. The chapter provides an overview of materials and explains the optimization of laser alignment.

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