小柯机器人

美国加州大学圣迭戈分校Lingyan Shi小组开发出具有A-PoD的超分辨率SRS显微镜。2023年2月16日，《自然—方法学》杂志在线发表了这项成果。

为了实现超分辨受激拉曼散射（SRS）成像，研究人员人员开发了一种反卷积算法，其为基于适应性矩估计（Adam）优化的点化反卷积（A-PoD），并证明了其空间分辨率在单个脂滴（LD）的膜上低于59nm。研究人员将A-PoD应用于空间相关的多光子荧光成像和来自不同样品的氧化氘（D₂O）探测SRS（DO-SRS）成像，以比较细胞和亚细胞细胞器中蛋白质和脂质的纳米级分布。研究人员人员成功地用A-PoD耦合DO-SRS在LD中辨别出新合成的脂质。A-PoD增强DO-SRS成像方法还用于揭示果蝇大脑样本在不同饮食下的代谢变化。这种新方法使研究人员能够定量测量生物分子的纳米共定位和细胞器中的代谢动力学。

据介绍，SRS提供了具有高信噪比的代谢动力学成像能力。然而，它的空间分辨率受到成像物镜的数值孔径和分子散射截面的限制。

附：英文原文

Title: Super-resolution SRS microscopy with A-PoD

Author: Jang, Hongje, Li, Yajuan, Fung, Anthony A., Bagheri, Pegah, Hoang, Khang, Skowronska-Krawczyk, Dorota, Chen, Xiaoping, Wu, Jane Y., Bintu, Bogdan, Shi, Lingyan

Issue&Volume: 2023-02-16

Abstract: Stimulated Raman scattering (SRS) offers the ability to image metabolic dynamics with high signal-to-noise ratio. However, its spatial resolution is limited by the numerical aperture of the imaging objective and the scattering cross-section of molecules. To achieve super-resolved SRS imaging, we developed a deconvolution algorithm, adaptive moment estimation (Adam) optimization-based pointillism deconvolution (A-PoD) and demonstrated a spatial resolution of lower than 59nm on the membrane of a single lipid droplet (LD). We applied A-PoD to spatially correlated multiphoton fluorescence imaging and deuterium oxide (D2O)-probed SRS (DO-SRS) imaging from diverse samples to compare nanoscopic distributions of proteins and lipids in cells and subcellular organelles. We successfully differentiated newly synthesized lipids in LDs using A-PoD-coupled DO-SRS. The A-PoD-enhanced DO-SRS imaging method was also applied to reveal metabolic changes in brain samples from Drosophila on different diets. This new approach allows us to quantitatively measure the nanoscopic colocalization of biomolecules and metabolic dynamics in organelles.

DOI: 10.1038/s41592-023-01779-1

Source: https://www.nature.com/articles/s41592-023-01779-1

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex