Third Institute of Physics - Biophysics, Georg August University, Friedrich-Hund-Platz 1, 37077 Göttingen, Germany.
Max Planck Institute for Dynamics and Self-Organization, Am Faßberg 17, 37077 Göttingen, Germany.
ACS Nano. 2023 May 9;17(9):8242-8251. doi: 10.1021/acsnano.2c12372. Epub 2023 Mar 30.
Metal-induced energy transfer (MIET) imaging is an easy-to-implement super-resolution modality that achieves nanometer resolution along the optical axis of a microscope. Although its capability in numerous biological and biophysical studies has been demonstrated, its implementation for live-cell imaging with fluorescent proteins is still lacking. Here, we present its applicability and capabilities for live-cell imaging with fluorescent proteins in diverse cell types (adult human stem cells, human osteo-sarcoma cells, and cells), and with various fluorescent proteins (GFP, mScarlet, RFP, YPet). We show that MIET imaging achieves nanometer axial mapping of living cellular and subcellular components across multiple time scales, from a few milliseconds to hours, with negligible phototoxic effects.
金属诱导能量转移(MIET)成像是一种易于实现的超分辨率模式,可沿着显微镜的光轴实现纳米级分辨率。尽管已经证明了它在许多生物学和生物物理学研究中的能力,但它在带有荧光蛋白的活细胞成像中的应用仍然缺乏。在这里,我们展示了它在不同细胞类型(成人干细胞、人骨肉瘤细胞和 细胞)以及各种荧光蛋白(GFP、mScarlet、RFP、YPet)中的应用和能力,用于活细胞成像。我们表明,MIET 成像可以在从几毫秒到几小时的多个时间尺度上实现纳米级的活细胞和亚细胞成分的轴向映射,而且几乎没有光毒性。