Bhaskar Haresh, Gidden Zoe, Virdi Gurvir, Kleinjan Dirk-Jan, Rosser Susan J, Gandhi Sonia, Regan Lynne, Horrocks Mathew H
School of Biological Sciences, The University of Edinburgh, Edinburgh, UK.
IRR Chemistry Hub, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh, UK.
Protein Sci. 2025 Feb;34(2):e70008. doi: 10.1002/pro.70008.
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.
超分辨率显微镜彻底改变了生物成像技术，能够在纳米长度尺度上可视化结构。然而，其在活细胞中的应用仍然具有挑战性。为了解决这个问题，我们将在酵母中建立的LIVE-PAINT方法进行了改进，以应用于活的哺乳动物细胞。使用101A/101B卷曲螺旋肽对作为基于肽的靶向系统，我们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，一种定位于细胞核，另一种定位于细胞质。这项研究突出了LIVE-PAINT的多功能性，表明其在哺乳动物细胞中对一系列蛋白质靶点进行活细胞超分辨率成像的潜力。我们将原始方法的哺乳动物细胞版本命名为mLIVE-PAINT。
