高通量成像阵列的荧光标记酵母突变株。
High-Throughput Imaging of Arrays of Fluorescently Tagged Yeast Mutant Strains.
机构信息
The Donnelly Centre, University of Toronto, Toronto, ON, Canada.
Department of Chemistry and Biology, Ryerson University, Toronto, ON, Canada.
出版信息
Methods Mol Biol. 2021;2304:221-242. doi: 10.1007/978-1-0716-1402-0_12.
Abstract
We describe a protocol for live-cell high-throughput (HTP) screening of yeast mutant strains carrying fluorescent protein markers for subcellular compartments of choice using automated confocal microscopy. This procedure, which combines HTP genetics and microscopy, results in the acquisition of thousands of images that can be analyzed in a systematic and quantitative way to identify morphology defects in the tagged subcellular compartments. This HTP protocol is readily adapted for screening any combination of markers and can be expanded to different growth conditions or higher order mutant genetic backgrounds.
摘要
我们描述了一种使用自动化共聚焦显微镜对携带荧光蛋白标记物的酵母突变株进行活细胞高通量（HTP）筛选的方案，这些标记物用于选择的细胞内区室。该方法结合了 HTP 遗传学和显微镜技术，可以获取数千张图像，然后以系统和定量的方式对这些图像进行分析，以鉴定标记的细胞内区室的形态缺陷。这种 HTP 方案易于适应任何标记物组合的筛选，并可扩展到不同的生长条件或更高阶的突变遗传背景。
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