Protocol to assess calcium signaling in response to metabolic alterations by flow cytometry and live-cell imaging.

Calcium (Ca) signaling is essential to cellular processes, such as signal transduction and migration, and dysregulation of Ca homeostasis can impact cellular function. Here, we present a protocol to analyze Ca signaling by its release into the cytosol in response to and after metabolic stimulation. We describe the steps for cell starvation from excessive nutrients and stimulation, followed by flow cytometry-based and microscopic analysis of calcium signaling. This protocol enables analysis of a large number of cells in a physiologically relevant microenvironment. For complete details on the use and execution of this protocol, please refer to Schlautmann et al..

Copyright © 2026 The Author(s). Published by Elsevier Inc. All rights reserved.

PMID: 41838718