Department of Electrical and Computer Engineering, University of California, San Diego, California.
Department of Neurosurgery, University of Minnesota, Minneapolis, Minnesota.
Cytometry A. 2019 May;95(5):499-509. doi: 10.1002/cyto.a.23764. Epub 2019 Apr 8.
Cell classification based on phenotypical, spatial, and genetic information greatly advances our understanding of the physiology and pathology of biological systems. Technologies derived from next generation sequencing and fluorescent activated cell sorting are cornerstones for cell- and genomic-based assays supporting cell classification and mapping. However, there exists a deficiency in technology space to rapidly isolate cells based on high content image information. Fluorescence-activated cell sorting can only resolve cell-to-cell variation in fluorescence and optical scattering. Utilizing microfluidics, photonics, computation microscopy, real-time image processing and machine learning, we demonstrate an image-guided cell sorting and classification system possessing the high throughput of flow cytometer and high information content of microscopy. We demonstrate the utility of this technology in cell sorting based on (1) nuclear localization of glucocorticoid receptors, (2) particle binding to the cell membrane, and (3) DNA damage induced γ-H2AX foci. © 2019 International Society for Advancement of Cytometry.
基于表型、空间和遗传信息的细胞分类极大地促进了我们对生物系统生理学和病理学的理解。源自下一代测序和荧光激活细胞分选的技术是支持细胞分类和绘图的基于细胞和基因组的检测的基石。然而,在基于高内涵图像信息快速分离细胞的技术方面存在不足。荧光激活细胞分选只能解析荧光和光散射的细胞间变化。我们利用微流控、光子学、计算显微镜、实时图像处理和机器学习,展示了一种具有流式细胞仪高通量和显微镜高信息量的图像引导细胞分选和分类系统。我们展示了该技术在基于以下方面的细胞分选方面的应用:(1)糖皮质激素受体的核定位,(2)颗粒与细胞膜的结合,以及(3)DNA 损伤诱导的 γ-H2AX 焦点。 © 2019 国际细胞分析协会。