Cells constantly generate reactive oxygen species (ROS) during aerobic metabolism. The ROS generation plays an important protective and functional role in the immune system. The cell is armed with a powerful antioxidant defense system to combat excessive production of ROS.

We present a detailed methodology to detect ROS generated as a result of specific FcγR stimulation beginning with macrophage generation, priming, staining, FcγR cross-linking, and ending with flow cytometric analysis.

MitoSOX-based flow cytometry is a quick, simple, and inexpensive method for detecting cellular ROS. If the probe is used at an optimal concentration, MitoSOX-based flow cytometry can reliably detect the relative differences in mitochondrial ROS formation in cells.

2019年3月7日 · We present a detailed methodology to detect ROS generated as a result of specific FcγR stimulation beginning with macrophage generation, priming, staining, FcγR cross-linking, and ending with flow cytometric analysis.

We describe a method for using multiparameter flow cytometry to detect mitochondrial reactive oxygen species (ROS) in murine healthy hematopoietic stem and progenitor cells (HSPCs) and leukemia cells from a mouse model of acute myeloid leukemia (AML) driven by MLL-AF9.

2010年1月1日 · Cells constantly generate reactive oxygen species (ROS) during aerobic metabolism. The ROS generation plays an important protective and functional role in the immune system. The cell is armed...

In this ROS Protocols article, we described the flow cytometric protocol involving the use of various concentrations of MitoSOX (1, 2.5, 5 μM) for detecting mitochondrial ROS in control and mitochondrial DNA-deficient (MD) melanoma B16-F10 cells.

2024年11月25日 · Flow cytometry is a powerful tool for measuring intracellular or mitochondrial ROS levels in real time. The following is a step-by-step guide to detecting ROS using flow cytometry: 1. Sample...

The detection of reactive oxygen species (ROS) and the analysis of oxidative stress are frequent applications of functional flow cytometry. Identifying and quantifying the ROS species generated during oxidative stress are crucial steps for the investigation of molecular mechanisms underlying stress responses.

2009年1月1日 · Flow cytometry or fluorimetry can be used for the detection of ROS with fluorescent probes. 2′-7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) is one of the most widely used techniques for directly measuring the redox state of a cell.