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Isolation of functional mitochondria by inertial microfluidics – a new method to sort intracellular organelles from a small scale biological sample

机译:惯性微流控技术分离功能性线粒体–一种从小规模生物样品中筛选细胞内细胞器的新方法

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Isolation of subcellular fractions from a limited amount of clinical sample is imperative for the continuous advancement of biological and clinical research. For example, analyses of pure and functional mitochondria from patient samples are expected not only to expand our knowledge towards the basic biological mechanisms of mitochondrial function, but also to elucidate how mitochondria are involved in the development of diseases such as cancers, premature aging syndromes, diabetes and neurodegenerative disorders. While currently available methods are mostly laborious and not suitable for small-scale analyses, we present a novel and facile approach to isolate mitochondria from limited amounts of biological samples by centrifugal-based differential migration. Furthermore, sorting cellular organelles by their intrinsic inertia enjoys the benefits of easy operation, undemanding equipment needs and continuous batch processing. Herein, we have successfully isolated functional mitochondria from crude cell lysate of less than 100 cells, which demonstrates the possibilities of promoting this methodology for detailed analyses of subcellular organelles, particularly when small-scale clinically relevant samples are considered.
机译:从数量有限的临床样品中分离亚细胞部分对于生物学和临床研究的不断发展是必不可少的。例如,从患者样本中分析纯净和功能性线粒体不仅有望将我们的知识扩展到线粒体功能的基本生物学机制,而且还将阐明线粒体如何参与疾病的发展,例如癌症,早衰综合症,糖尿病和神经退行性疾病。虽然当前可用的方法大多数是费力的并且不适用于小规模分析,但我们提出了一种新颖且简便的方法,可通过基于离心的差异迁移从有限量的生物样品中分离线粒体。此外,通过其固有惯性对细胞器进行分选具有操作简便,设备需求不高以及连续批量处理的优点。在这里,我们已经成功地从少于100个细胞的粗细胞裂解物中分离出功能性线粒体,这证明了推广此方法用于亚细胞细胞器详细分析的方法的可能性,尤其是在考虑到小规模临床相关样品时。

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