小柯机器人

瑞典皇家理工学院的kIlaria Testa团队使用三维平行RESOLFT显微镜实现活细胞体积成像。相关论文于2021年1月11日在线发表于国际学术期刊《自然—生物技术》。

研究人员报道了一种3D、平行、可逆、可饱和/可切换的光学荧光跃迁（3D pRESOLFT）显微镜，其能够在整个活细胞中提供低于80 nm的3D分辨率。通过具有干扰图案的高度平行图像采集，研究人员实现了大视野（约40×40 µm²）的快速（1-2 Hz）采集，从而创建了3D限制且等距间隔的强度最小值阵列。这使得研究人员能够将可切换的荧光蛋白可逆地转换为暗态，从而实现了有针对性的3D荧光限制。

研究人员在培养的海马神经元可塑性期间，可视化了活细胞中细胞器的3D组织和动态以及突触的体积结构变化。

据了解，阐明细胞内细胞器和分子的体积结构需要在所有三个维度上具有足够高的空间分辨率。当前的方法受到沿光轴的分辨能力不足、记录时间长以及应用于活细胞成像时光漂白的限制。

附：英文原文

Title: Volumetric live cell imaging with three-dimensional parallelized RESOLFT microscopy

Author: Andreas Bodn, Francesca Pennacchietti, Giovanna Coceano, Martina Damenti, Michael Ratz, Ilaria Testa

Issue&Volume: 2021-01-11

Abstract: Elucidating the volumetric architecture of organelles and molecules inside cells requires microscopy methods with a sufficiently high spatial resolution in all three dimensions. Current methods are limited by insufficient resolving power along the optical axis, long recording times and photobleaching when applied to live cell imaging. Here, we present a 3D, parallelized, reversible, saturable/switchable optical fluorescence transition (3D pRESOLFT) microscope capable of delivering sub-80-nm 3D resolution in whole living cells. We achieved rapid (1–2Hz) acquisition of large fields of view (~40×40μm2) by highly parallelized image acquisition with an interference pattern that creates an array of 3D-confined and equally spaced intensity minima. This allowed us to reversibly turn switchable fluorescent proteins to dark states, leading to a targeted 3D confinement of fluorescence. We visualized the 3D organization and dynamics of organelles in living cells and volumetric structural alterations of synapses during plasticity in cultured hippocampal neurons.

DOI: 10.1038/s41587-020-00779-2

Source: https://www.nature.com/articles/s41587-020-00779-2

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex