{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T02:56:56Z","timestamp":1781837816910,"version":"3.54.5"},"reference-count":18,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,3,6]],"date-time":"2023-03-06T00:00:00Z","timestamp":1678060800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Biom\u00e9decine AOR Greffe 2017","doi-asserted-by":"publisher","award":["ANR-18-CE18-0015"],"award-info":[{"award-number":["ANR-18-CE18-0015"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001665","name":"Agence Biom\u00e9decine AOR Greffe 2017","doi-asserted-by":"publisher","award":["PME20170637799"],"award-info":[{"award-number":["PME20170637799"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"name":"FRM","award":["ANR-18-CE18-0015"],"award-info":[{"award-number":["ANR-18-CE18-0015"]}]},{"name":"FRM","award":["PME20170637799"],"award-info":[{"award-number":["PME20170637799"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Rationale: Mitochondria are key organelles involved in cell survival and death during the acute phenomena of myocardial ischemia-reperfusion (i.e., myocardial infarction). To investigate the functions of isolated mitochondria such as calcium retention capacity, oxidative phosphorylation, and reactive oxygen species (ROS) production, already established methods are based on extramitochondrial measurements of the whole mitochondria population. Objective: The aim of this study was to develop a reliable and well-characterized method for multiparametric analysis of isolated single mitochondrion by flow cytometry (FC) in the context of myocardial infarction. The advantage of FC is the possibility to give a simultaneous analysis of morphological parameters (side and forward scatters: SSC and FSC) for each mitochondrion, combined with intramitochondrial measurements of several biological markers, such as ROS production or membrane potential (\u0394\u03c6m), using specific fluorescent probes. Methods and Results: For this study, a rat model of ischemia-reperfusion and a protective approach of post-conditioning using low reperfusion pressure was used. Thanks to the use of specific probes (NAO, MTR, TMRM, DilC1, and DHR123) combined with flow cytometry, we propose a method: (i) to identify mitochondrial populations of interest based on quality criteria (NAO\/TMRM double staining); (ii) to monitor their morphological criteria, especially during swelling due to calcium overload; and (iii) to compare mitochondrial functions (membrane potential and ROS production) in different experimental groups. Applied to mitochondria from ischemic hearts, these measurements revealed that individual mitochondria are altered and that cardioprotection by low-pressure reperfusion reduces damage, as expected. Conclusions: Our results highlight FC as a reliable and sensitive method to investigate changes in mitochondrial functions and morphology in pathological conditions that disrupts their activity such as the case in ischemia-reperfusion. This methodological approach can be extended to other pathologies involving mitochondrial dysfunctions. Moreover, FC offers the possibility to work with very small amounts of isolated mitochondria, a factor that may limit the use of classical methods.<\/jats:p>","DOI":"10.3390\/life13030707","type":"journal-article","created":{"date-parts":[[2023,3,6]],"date-time":"2023-03-06T05:18:43Z","timestamp":1678079923000},"page":"707","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Isolated Mitochondria State after Myocardial Ischemia-Reperfusion Injury and Cardioprotection: Analysis by Flow Cytometry"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2200-9511","authenticated-orcid":false,"given":"Claire","family":"Crola Da Silva","sequence":"first","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7678-062X","authenticated-orcid":false,"given":"Delphine","family":"Baetz","sequence":"additional","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"V\u00e9d\u00e8re","sequence":"additional","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e9gane","family":"Lo-Grasso","sequence":"additional","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6764-5351","authenticated-orcid":false,"given":"Mariam","family":"Wehbi","sequence":"additional","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christophe","family":"Chouabe","sequence":"additional","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel","family":"Bidaux","sequence":"additional","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7643-7784","authenticated-orcid":false,"given":"Ren\u00e9","family":"Ferrera","sequence":"additional","affiliation":[{"name":"University of Lyon, CARMEN Laboratory, INSERM, INRAE, Universit\u00e9 Claude Bernard Lyon 1, 69500 Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1097\/MOT.0000000000001015","article-title":"Mitochondria and ischemia reperfusion injury","volume":"27","author":"Panconesi","year":"2022","journal-title":"Curr. Opin. Organ. Transplant."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Ferrera, R., V\u00e9d\u00e8re, M., Lo-Grasso, M., Augeul, L., Chouabe, C., Bidaux, G., and Baetz, D. (2022). Postconditioning by Delayed Administration of Ciclosporin A: Implication for Donation after Circulatory Death (DCD). Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232112858"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Stevic, N., Maalouf, J., Argaud, L., Gallo-Bona, N., Lo Grasso, M., Gouriou, Y., Gomez, L., Crola Da Silva, C., Ferrera, R., and Ovize, M. (2022). Cooling Uncouples Differentially ROS Production from Respiration and Ca2+Homeostasis Dynamic in Brain and Heart Mitochondria. Cells, 11.","DOI":"10.3390\/cells11060989"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1146\/annurev-anchem-061020-111722","article-title":"Analysis of Mitochondria by Single-Organelle Resolution","volume":"15","author":"Harbauer","year":"2022","journal-title":"Annu. 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