德国慕尼黑工业大学Gil Gregor Westmeyer团队近期取得重要工作进展，他们研究开发出了用于相关体积电子显微镜的基因编码条形码。相关研究成果2023年4月17日在线发表于《自然—生物技术》杂志上。
据介绍，虽然报告基因在荧光显微镜中很常见，但用于电子显微镜（EM）的等效多路复用报告基因仍然很少。
通过在不同尺寸的囊泡纳米室的管腔中安装可变数量的固定稳定的金属相互作用部分，研究人员开发了一套可通过标准EM技术读取的球对称和同心条形码（EMcapsulins）。六类EMcapsulins可以自动分割和区分。通过一组可变长度的刚性间隔物将几个EMcapsulins排列成不同的模式，能进一步提高信息编码能力。荧光EMcapsulins被表达以监测光和EM中的亚细胞结构。果蝇和小鼠大脑中的神经元表达使EM中的基因定义细胞能够自动识别。EMcapsulins与透射EM、扫描EM和聚焦离子束扫描EM兼容。
总之，基因控制的EM可读条形码的可扩展调色板可以用多路复用的基因表达图来增强解剖EM图像。
附：英文原文
Title: Genetically encoded barcodes for correlative volume electron microscopy
Author: Sigmund, Felix, Berezin, Oleksandr, Beliakova, Sofia, Magerl, Bernhard, Drawitsch, Martin, Piovesan, Alberto, Gonalves, Filipa, Bodea, Silviu-Vasile, Winkler, Stefanie, Bousraou, Zoe, Grosshauser, Martin, Samara, Eleni, Pujol-Mart, Jess, Schdler, Sebastian, So, Chun, Irsen, Stephan, Walch, Axel, Kofler, Florian, Piraud, Marie, Kornfeld, Joergen, Briggman, Kevin, Westmeyer, Gil Gregor
Issue&Volume: 2023-04-17
Abstract: While genetically encoded reporters are common for fluorescence microscopy, equivalent multiplexable gene reporters for electron microscopy (EM) are still scarce. Here, by installing a variable number of fixation-stable metal-interacting moieties in the lumen of encapsulin nanocompartments of different sizes, we developed a suite of spherically symmetric and concentric barcodes (EMcapsulins) that are readable by standard EM techniques. Six classes of EMcapsulins could be automatically segmented and differentiated. The coding capacity was further increased by arranging several EMcapsulins into distinct patterns via a set of rigid spacers of variable length. Fluorescent EMcapsulins were expressed to monitor subcellular structures in light and EM. Neuronal expression in Drosophila and mouse brains enabled the automatic identification of genetically defined cells in EM. EMcapsulins are compatible with transmission EM, scanning EM and focused ion beam scanning EM. The expandable palette of genetically controlled EM-readable barcodes can augment anatomical EM images with multiplexed gene expression maps.
DOI: 10.1038/s41587-023-01713-y
Source: https://www.nature.com/articles/s41587-023-01713-y
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
本期文章：《自然—生物技术》：Online/在线发表
