A novel and effective separation method for single mitochondria analysis |
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| Authors: | René Pflugradt Ulrike Schmidt Benjamin Landenberger Timo Sänger Sabine Lutz-Bonengel |
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| Affiliation: | 1. Department of Neurology, University of Halle-Wittenberg, Ernst-Grube-Str. 40, Halle/Saale 06097, Germany;2. Wellcome Trust Centre for Mitochondrial Research, Institute of Neuroscience, The Medical School, Newcastle University, Framlington Place, Newcastle upon Tyne, UK;1. Biology Department, Portland State University, 1719 SW 10th Ave., Portland, OR 97201, USA;2. Department of Zoology, Oregon State University, Corvallis, OR 97331, USA;3. Center for Genome Research and Biocomputing, Oregon State University, Corvallis, OR 97331, USA;1. Wellcome Trust Centre for Mitochondrial Research, Institute for Ageing and Health, Newcastle University, Newcastle upon Tyne, United Kingdom;2. Salford Royal NHS Foundation Trust, Greater Manchester Neuroscience Centre, Department of Neurology, Salford, Lanchester, M6 8HD, United Kingdom |
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| Abstract: | To investigate the set of mtDNA molecules contained in small biological structures, powerful techniques for separation are required. We tested flow cytometry (FCM1), laser capture microdissection (LCM2) and a method using optical tweezers (OT3) in combination with a 1μ-Ibidi-Slide with regard to their ability to deposit single mitochondrial particles. The success of separation was determined by real-time quantitative PCR (qPCR4) and sequencing analysis.OT revealed the highest potential for the separation and deposition of single mitochondrial particles. The study presents a novel setup for effective separation of single mitochondrial particles, which is crucial for the analysis of single mitochondria. |
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