Landry Zachary C, Giovanonni Stephen J, Quake Stephen R, Blainey Paul C
Department of Microbiology, Oregon State University, Corvallis, Oregon, USA.
Methods Enzymol. 2013;531:61-90. doi: 10.1016/B978-0-12-407863-5.00004-6.
Genetic analysis of single cells is emerging as a powerful approach for studies of heterogeneous cell populations. Indeed, the notion of homogeneous cell populations is receding as approaches to resolve genetic and phenotypic variation between single cells are applied throughout the life sciences. A key step in single-cell genomic analysis today is the physical isolation of individual cells from heterogeneous populations, particularly microbial populations, which often exhibit high diversity. Here, we detail the construction and use of instrumentation for optical trapping inside microfluidic devices to select individual cells for analysis by methods including nucleic acid sequencing. This approach has unique advantages for analyses of rare community members, cells with irregular morphologies, small quantity samples, and studies that employ advanced optical microscopy.
单细胞的遗传分析正在成为研究异质细胞群体的一种强大方法。事实上，随着在整个生命科学领域应用解决单细胞之间遗传和表型变异的方法，同质细胞群体的概念正在逐渐淡化。当今单细胞基因组分析的一个关键步骤是从异质群体，特别是微生物群体中物理分离单个细胞，这些群体通常表现出高度的多样性。在这里，我们详细介绍了微流控装置内用于光镊的仪器的构建和使用，以通过包括核酸测序在内的方法选择单个细胞进行分析。这种方法对于分析稀有群落成员、形态不规则的细胞、少量样本以及采用先进光学显微镜的研究具有独特优势。
