标准IgG抗体助力小鼠全身细胞定位

德国慕尼黑亥姆霍兹研究中心Ali Ertürk研究小组宣布他们使用标准IgG抗体在小鼠中完成了全身细胞定位。该研究于2023年7月10日发表于国际学术期刊《自然—生物技术》杂志。

研究人员研发了wildDISCO，这是一种用于小鼠全身免疫标记、光学清除和成像的新方法，其消除了对转基因报告动物或纳米抗体标记的需求，从而克服了现有的技术壁垒。研究人员发现庚基（2，6-二-O-甲基）-β-环糊精可作为胆固醇提取和膜透化的强效增强剂，能够深度、均匀地渗透标准抗体而不会产生聚集。WildDISCO通过标记不同的内源性蛋白质，实现在细胞分辨率水平对小鼠周围神经系统、淋巴管和免疫细胞的成像。

此外，研究还探索了罕见增殖细胞和生物扰动的影响，与通过无菌小鼠所证明的结果一致。研究人员应用wildDISCO对乳腺癌小鼠的三级淋巴结构进行了绘制，同时囊括了原发性肿瘤和整个小鼠体内的转移情况。可以通过访问http://discotechnologies.org/wildDISCO/atlas/index.php获得小鼠神经、淋巴和血管系统的高分辨率图像图集。

据了解，全身成像技术在研究生理系统对健康维持和疾病产生的相互作用中发挥着至关重要的作用。

附：英文原文

Title: Whole-body cellular mapping in mouse using standard IgG antibodies

Author: Mai, Hongcheng, Luo, Jie, Hoeher, Luciano, Al-Maskari, Rami, Horvath, Izabela, Chen, Ying, Kofler, Florian, Piraud, Marie, Paetzold, Johannes C., Modamio, Jennifer, Todorov, Mihail, Elsner, Markus, Hellal, Farida, Ertrk, Ali

Issue&Volume: 2023-07-10

Abstract: Whole-body imaging techniques play a vital role in exploring the interplay of physiological systems in maintaining health and driving disease. We introduce wildDISCO, a new approach for whole-body immunolabeling, optical clearing and imaging in mice, circumventing the need for transgenic reporter animals or nanobody labeling and so overcoming existing technical limitations. We identified heptakis(2,6-di-O-methyl)-β-cyclodextrin as a potent enhancer of cholesterol extraction and membrane permeabilization, enabling deep, homogeneous penetration of standard antibodies without aggregation. WildDISCO facilitates imaging of peripheral nervous systems, lymphatic vessels and immune cells in whole mice at cellular resolution by labeling diverse endogenous proteins. Additionally, we examined rare proliferating cells and the effects of biological perturbations, as demonstrated in germ-free mice. We applied wildDISCO to map tertiary lymphoid structures in the context of breast cancer, considering both primary tumor and metastases throughout the mouse body. An atlas of high-resolution images showcasing mouse nervous, lymphatic and vascular systems is accessible at http://discotechnologies.org/wildDISCO/atlas/index.php.

DOI: 10.1038/s41587-023-01846-0

Source: https://www.nature.com/articles/s41587-023-01846-0