A nanoscale, multi-parametric flow cytometry-based platform to study mitochondrial heterogeneity and mitochondrial DNA dynamics.
Julie A MacDonaldAlisha M BothunSofia N AnnisHannah SheehanSomak RayYuanwei GaoAlexander R IvanovKonstantin KhrapkoJonathan L TillyDori C WoodsPublished in: Communications biology (2019)
Mitochondria are well-characterized regarding their function in both energy production and regulation of cell death; however, the heterogeneity that exists within mitochondrial populations is poorly understood. Typically analyzed as pooled samples comprised of millions of individual mitochondria, there is little information regarding potentially different functionality across subpopulations of mitochondria. Herein we present a new methodology to analyze mitochondria as individual components of a complex and heterogeneous network, using a nanoscale and multi-parametric flow cytometry-based platform. We validate the platform using multiple downstream assays, including electron microscopy, ATP generation, quantitative mass-spectrometry proteomic profiling, and mtDNA analysis at the level of single organelles. These strategies allow robust analysis and isolation of mitochondrial subpopulations to more broadly elucidate the underlying complexities of mitochondria as these organelles function collectively within a cell.
Keyphrases
- cell death
- flow cytometry
- mitochondrial dna
- single cell
- high throughput
- copy number
- oxidative stress
- reactive oxygen species
- endoplasmic reticulum
- mass spectrometry
- electron microscopy
- cell cycle arrest
- high resolution
- randomized controlled trial
- gene expression
- social media
- gas chromatography
- mesenchymal stem cells
- simultaneous determination
- data analysis