Super-resolution imaging of proteins inside live mammalian cells with mLIVE-PAINT

Haresh Bhaskar, Zoe Gidden, Gurvir Virdi, Dirk-Jan Kleinjan, Susan J. Rosser, Sonia Gandhi, Lynne Regan, Mathew H. Horrocks
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Abstract

Super-resolution microscopy has revolutionized biological imaging, enabling the visualization of structures at the nanometer length scale. Its application in live cells, however, has remained challenging. To address this, we adapted LIVE-PAINT, an approach we established in yeast, for application in live mammalian cells. Using the 101A/101B coiled-coil peptide pair as a peptide-based targeting system, we successfully demonstrate the super-resolution imaging of two distinct proteins in mammalian cells, one localized in the nucleus, and the second in the cytoplasm. This study highlights the versatility of LIVE-PAINT, suggesting its potential for live-cell super-resolution imaging across a range of protein targets in mammalian cells. We name the mammalian cell version of our original method mLIVE-PAINT.
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利用 mLIVE-PAINT 对哺乳动物活细胞内的蛋白质进行超分辨率成像
超分辨显微镜彻底改变了生物成像技术,使纳米级的结构可视化成为可能。然而,超分辨显微镜在活细胞中的应用仍具有挑战性。为了解决这个问题,我们将在酵母中建立的 LIVE-PAINT 方法应用于活哺乳动物细胞。我们使用 101A/101B 盘卷肽对作为基于肽的靶向系统,成功地展示了哺乳动物细胞中两种不同蛋白质的超分辨率成像,其中一种定位在细胞核中,另一种定位在细胞质中。这项研究凸显了 LIVE-PAINT 的多功能性,表明它有潜力对哺乳动物细胞中的一系列蛋白质靶标进行活细胞超分辨率成像。我们将这种哺乳动物细胞版本的原始方法命名为 mLIVE-PAINT。
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