Protocol for investigating mitochondrial structure, function, and metabolism in human cervical cancer cells
Year: 2025
Type: article
Abstract: Mitochondria regulate a variety of biological activities, including metabolism, oxidative stress, and cell death. Here, we present a protocol for the investigation of mitochondrial structure, function, and metabolism in human cervical cancer cells. We describe steps for staining and visualizing mitochondria using confocal microscopy to assess morphology, mass, membrane potential, calcium, reactive oxygen species (ROS), and lipid droplet accumulation. We then detail procedures for isolating mitochondria and performing metabolomic analysis of mitochondrial metabolites via mass spectrometry. For complete details on the use and execution of this protocol, please refer to Adiga et al. 1 • Protocol to study mitochondrial structure, function, and metabolism comprehensively • Stepwise protocol for mitochondrial staining using confocal microscopy • Modular staining for multi-parameter assays • Mass spectrometry-based metabolomic analysis of mitochondrial metabolites Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Mitochondria regulate a variety of biological activities, including metabolism, oxidative stress, and cell death. Here, we present a protocol for the investigation of mitochondrial structure, function, and metabolism in human cervical cancer cells. We describe steps for staining and visualizing mitochondria using confocal microscopy to assess morphology, mass, membrane potential, calcium, reactive oxygen species (ROS), and lipid droplet accumulation. We then detail procedures for isolating mitochondria and performing metabolomic analysis of mitochondrial metabolites via mass spectrometry.
Source: STAR Protocols
Authors: Sachin Shetty, Shama Prasada Kabekkodu
Institution: Manipal Academy of Higher Education
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W4416437679