biostudies-literatureUnknown1(1)Bertan FInnovative Medicines InitiativeDFGHorizon 2020 Marie Skłodowska-Curie ActionsMitochondria sustain the energy demand of the cell. The composition and functional state of the mitochondrial oxidative phosphorylation system are informative indicators of organelle bioenergetic capacity. Here, we describe a highly sensitive and reproducible method for a single-cell quantification of mitochondrial CI- and CIV-containing respiratory supercomplexes (CI∗CIV-SCs) as an alternative means of assessing mitochondrial respiratory chain integrity. We apply a proximity ligation assay (PLA) and stain CI∗CIV-SCs in fixed human and mouse brains, tumorigenic cells, induced pluripotent stem cells (iPSCs) and iPSC-derived neural precursor cells (NPCs), and neurons. Spatial visualization of CI∗CIV-SCs enables the detection of mitochondrial lesions in various experimental models, including Cell reports methods100002https://www.ebi.ac.uk/biostudies/studies/S-EPMC9017192biostudies-literatureComparative analysis of CI- and CIV-containing respiratory supercomplexes at single-cell resolution.PMC9017192Bertan FMarsal-Cots ANicotera PScifo EPrehn JHMPeitz MBano DEhninger DGuranda MPiazzesi AWischhof LStork MJackson JfalseComparative analysis of CI- and CIV-containing respiratory supercomplexes at single-cell resolution.Mitochondria sustain the energy demand of the cell. The composition and functional state of the mitochondrial oxidative phosphorylation system are informative indicators of organelle bioenergetic capacity. Here, we describe a highly sensitive and reproducible method for a single-cell quantification of mitochondrial CI- and CIV-containing respiratory supercomplexes (CI∗CIV-SCs) as an alternative means of assessing mitochondrial respiratory chain integrity. We apply a proximity ligation assay (PLA) and stain CI∗CIV-SCs in fixed human and mouse brains, tumorigenic cells, induced pluripotent stem cells (iPSCs) and iPSC-derived neural precursor cells (NPCs), and neurons. Spatial visualization of CI∗CIV-SCs enables the detection of mitochondrial lesions in various experimental models, including 2021-01-01T00:00:00Z2021 May2026-05-09T21:52:01.982Z2024-11-20T22:37:38.162ZS-EPMC90171923547469410.1016/j.crmeth.2021.100002