Improved spatial resolution by induced live cell and organelle swelling in hypotonic solutions
Improved spatial resolution by induced live cell and organelle swelling in hypotonic solutions
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DOI：
10.1038/s41598-019-49408-2
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发表时间：
2019-09-09
影响因子：
4.6
通讯作者：
Lorenz, Holger
中科院分区：
文献类型：
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作者：
Jaiswal, Astha;Hoerth, Christian H.;Lorenz, Holger
诱导细胞和细胞器的形态变化是迄今为止在光学显微镜中确定精确蛋白质亚位置和细胞器数量的最简单方法。通过使用低渗溶液来溶胀哺乳动物细胞器，我们证明了可以可靠地建立内质网、高尔基体和线粒体内的精确膜、腔或基质蛋白位置。我们还显示了这种方法对细胞器定量的好处，特别是对于聚集或缠绕的细胞器，如过氧化物酶体和线粒体。由于细胞和细胞器的肿胀是可逆的，它可以应用于活细胞连续的高分辨率分析。我们的方法优于许多现有的成像模式的分辨率，易用性和成本效益，而不排除任何与现有的光学（超）分辨率技术的共同利用。
Induced morphology changes of cells and organelles are by far the easiest way to determine precise protein sub-locations and organelle quantities in light microscopy. By using hypotonic solutions to swell mammalian cell organelles we demonstrate that precise membrane, lumen or matrix protein locations within the endoplasmic reticulum, Golgi and mitochondria can reliably be established. We also show the benefit of this approach for organelle quantifications, especially for clumped or intertwined organelles like peroxisomes and mitochondria. Since cell and organelle swelling is reversible, it can be applied to live cells for successive high-resolution analyses. Our approach outperforms many existing imaging modalities with respect to resolution, ease-of-use and cost-effectiveness without excluding any co-utilization with existing optical (super) resolution techniques.