Ramunas John, Illman Meredith, Kam Angela, Farn Kristen, Kelly Liam, Morshead Cindi M, Jervis Eric J
Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
Cytometry A. 2006 Dec 1;69(12):1202-11. doi: 10.1002/cyto.a.20341.
There is a need for methods to (1) track cells continuously to generate lineage trees; (2) culture cells in in vivo-like microenvironments; and (3) measure many biological parameters simultaneously and noninvasively. Herein, we present a novel imaging culture chamber that facilitates "lineage informatics," a lineage-centric approach to cytomics.
We cultured cells in a confined monolayer using a novel "gap chamber" that produces images with confocal-like qualities using standard DIC microscopy. Lineage and other cytometric data were semiautomatically extracted from image sets of neural stem and progenitor cells and analyzed using lineage informatics.
Cells imaged in the chamber every 3 min could be tracked for at least 6 generations allowing for the construction of extensive lineage trees with multiparameter data sets at hundreds of time points for each cell. The lineage informatics approach reveals relationships between lineage, phenotype, and microenvironment. Mass transfer characteristics and 3D geometry make the chamber more in vivo-like than traditional culture systems.
The gap chamber allows cells to be cultured, imaged, and tracked in true monolayers permitting detailed informatics analysis of cell lineage, phenotype, and fate determinants. The chamber is biomimetic and straightforward to build and use, and should find many applications in long-term cell imaging.
需要有方法来（1）持续追踪细胞以生成谱系树；（2）在类似体内的微环境中培养细胞；以及（3）同时且无创地测量多种生物学参数。在此，我们展示了一种新型成像培养室，它有助于“谱系信息学”，这是一种以谱系为中心的细胞组学方法。
我们使用一种新型“间隙室”在受限单层中培养细胞，该间隙室使用标准微分干涉差显微镜产生具有共聚焦样质量的图像。从神经干细胞和祖细胞的图像集中半自动提取谱系和其他细胞测量数据，并使用谱系信息学进行分析。
每3分钟在培养室中成像的细胞可被追踪至少6代，从而能够构建广泛的谱系树，每个细胞在数百个时间点具有多参数数据集。谱系信息学方法揭示了谱系、表型和微环境之间的关系。传质特性和三维几何结构使该培养室比传统培养系统更类似体内环境。
间隙室允许细胞在真正的单层中进行培养、成像和追踪，从而对细胞谱系、表型和命运决定因素进行详细的信息学分析。该培养室具有仿生特性，构建和使用都很简单，并且在长期细胞成像中应有许多应用。
