2023-07-11 ミュンヘン大学(LMU)
◆wildDISCOのワークフローは、血管内注入による全身への抗体の分布、動物の光学的クリアリング、ライトシート蛍光顕微鏡法を含みます。wildDISCOを使用することで、研究者たちは、神経系、リンパ管、血管、免疫細胞に焦点を当てたマウス全身アトラスを作成することができました。
<関連情報>
- https://www.lmu.de/en/newsroom/news-overview/news/wilddisco-visualizing-whole-bodies-in-unprecedented-cetail.html
- https://www.nature.com/articles/s41587-023-01846-0
標準IgG抗体を用いたマウスの全身細胞マッピング Whole-body cellular mapping in mouse using standard IgG antibodies
Hongcheng Mai,Jie Luo,Luciano Hoeher,Rami Al-Maskari,Izabela Horvath,Ying Chen,Florian Kofler,Marie Piraud,Johannes C. Paetzold,Jennifer Modamio,Mihail Todorov,Markus Elsner,Farida Hellal & Ali Ertürk
Nature Biotechnology Published:10 July 2023
DOI:https://doi.org/10.1038/s41587-023-01846-0
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.
