Abstract
The understanding of how different cell types adapt their metabolism in the face of challenges has been attracting the attention of researchers for many years. Recently, immunologists also started to focus on how the metabolism of immune cells can impact the way that immunity drives its responses. The presence of a pathogen or damage in a tissue changes severely the way that the immune cells need to respond. When activated, immune cells usually shift their metabolism from a high energy demanding status using mitochondria respiration to a glycolytic based rapid ATP production. The diminished amount of respiration leads to changes in the mitochondrial membrane potential and, consequently, generation of reactive oxygen species. Here, we show how flow cytometry can be used to track changes in mitochondrial mass, membrane potential and superoxide (ROS) production in live immune cells.● This protocol suggests a quick way of evaluating mitochondrial fitness using flow cytometry. We propose using the probes MitoTraker Green and MitoTracker Red/ MitoSOX at the same time. This way, it is possible to evaluate different parameters of mitochondrial biology in living cells.● Flow cytometry is a highly used tool by immunologists. With the advances of studies focusing on the metabolism of immune cells, a simplified application of flow cytometry for mitochondrial studies and screenings is a helpful clarifying method for immunology.
Highlights
The metabolism of a cell is an intriguing dynamic workflow of metabolites and enzymes that adapt to the needs of each cell in a certain environment
Immunology and Microbiology Immunometabolism Flow cytometry – Mitochondrial staining Cell staining with mitochondrial probes (MitoTracker Green, Red and MitoSOX) and acquisition in flow cytometer has been published by Hagai Rottenberg and ShaolongWu. (1998)
We describe how mitochondrial specific dyes can be used to track mitochondrial function in a simple flow cytometry assay
Summary
Lauar de Brito Monteiro a, Gustavo Gastão Davanzo a, Cristhiane Favero de Aguiar a, Pedro M.M. We show how flow cytometry can be used to track changes in mitochondrial mass, membrane potential and superoxide (ROS) production in live immune cells. We propose using the probes MitoTraker Green and MitoTracker Red/ MitoSOX at the same time This way, it is possible to evaluate different parameters of mitochondrial biology in living cells. Method name: Flow cytometry – Mitochondrial staining Keywords: Immunometabolism, Mitochondrial function, Mitochondrial membrane potential, Metabolic reprogramming Article history: Received 18 March 2020; Accepted 20 May 2020; Available online 28 May 2020. Immunology and Microbiology Immunometabolism Flow cytometry – Mitochondrial staining Cell staining with mitochondrial probes (MitoTracker Green, Red and MitoSOX) and acquisition in flow cytometer has been published by Hagai Rottenberg and ShaolongWu.
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