Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland.
Nat Methods. 2018 Jan;15(1):39-46. doi: 10.1038/nmeth.4503. Epub 2017 Nov 13.
Multicolor 3D quantitative imaging of large tissue volumes is necessary to understand tissue development and organization as well as interactions between distinct cell types in situ. However, tissue imaging remains technically challenging, particularly imaging of bone and marrow. Here, we describe a pipeline to reproducibly generate high-dimensional quantitative data from bone and bone marrow that may be extended to any tissue. We generate thick bone sections from adult mouse femurs with preserved tissue microarchitecture and demonstrate eight-color imaging using confocal microscopy without linear unmixing. We introduce XiT, an open-access software for fast and easy data curation, exploration and quantification of large imaging data sets with single-cell resolution. We describe how XiT can be used to correct for potential artifacts in quantitative 3D imaging, and we use the pipeline to measure the spatial relationship between hematopoietic cells, bone matrix and marrow Schwann cells.
多色 3D 定量成像大面积组织体积对于理解组织发育和组织以及不同细胞类型在体内的相互作用是必要的。然而，组织成像仍然具有技术挑战性，特别是骨和骨髓的成像。在这里，我们描述了一个可重复性地从骨和骨髓中生成高维定量数据的管道，该管道可以扩展到任何组织。我们从成年小鼠股骨中生成具有保留组织微观结构的厚骨切片，并展示了无需线性解混即可使用共聚焦显微镜进行的八色成像。我们引入了 XiT，这是一种用于快速轻松地管理、探索和量化具有单细胞分辨率的大型成像数据集的开源软件。我们描述了如何使用 XiT 来纠正定量 3D 成像中的潜在伪影，并且我们使用该管道来测量造血细胞、骨基质和骨髓施万细胞之间的空间关系。
