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Increased intracellular Ca2+ content is an essential and irreversible step towards the activation of T lymphocytes and indicated in regulating their cell survival, proliferation and differentiation. Upon antigen recognition, activation of the TCR initiates numerous signaling cascades resulting in cellular activation and proliferation. During TCR signaling, IL-2-inducible kinase (ITK) activates phospholipase C (PLC) and the subsequent release of intracellular Ca2+ stores from the endoplasmic reticulum (ER) resulting in a rapid elevation of intracellular Ca2+ levels up to 500nM. This process subsequently triggers the opening of ion channels in the plasma membrane allowing an influx of extracellular Ca2+, which in turn activates key molecules such as Calcineurin. Inhibition of calcium flux has been shown to affect T cell function, therefore, measuring the Ca2+ content in T cells presents important information about the status and kinetics of T cell activation. The capability of the NovoCyte flow cytometer to quantify dynamic changes in intracellular Ca2+ was demonstrated by measuring calcium flux in Jurkat T cells. This data demonstrates that dynamic alterations in intracellular Ca2+ concentration can be measured on the NovoCyte flow cytometer.
Related technologies: Conventional flow cytometry