Image-guided cell sorting using fast scanning lasers
Image-guided cell sorting using fast scanning lasers
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DOI：
10.1063/1.5144117
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发表时间：
2020-04-01
期刊：
影响因子：
5.6
通讯作者：
Lo, Yu-Hwa
中科院分区：
文献类型：
--
作者：
Chen, Xinyu;Gu, Yi;Lo, Yu-Hwa
除了传统的荧光标记生物标志物之外，根据细胞的成像和空间特征对细胞类型进行分类和分离靶向细胞，可以开发新的生物学见解，并建立表型和形态之间的联系，以及正常和病变状态下的基因组细胞信息。在这里，我们展示了一种微流控流式细胞仪细胞分选器，它使用快速扫描激光激发源和光电倍增管（pmt），结合实时图像处理，根据用户定义的空间特征对细胞进行成像和分选。荧光标记生物标志物的大小、形状、散射模式、形态等无标记特征以及空间分布等数十种特征极大地扩展了细胞分析中的可用信息空间。使用与成像系统兼容的一次性微流控墨盒有助于缩小尺寸并降低交叉污染的风险。所谓的无摄像头设计，使用pmt代替CCD，使系统更兼容传统的荧光激活细胞分选器和用户友好，同时享受pmt的灵敏度和好处。细胞图像特征可以直接从时间波形中提取，也可以通过时空变换从重建的细胞图像中提取。
Classification of cell types and isolation of targeted cells according to their imaging and spatial characteristics, beyond traditional fluorescently labeled biomarkers, enable the development of new biological insight and establishment of connections between phenotypical and morphological, and genomic cell information in normal and diseased states. Here, we demonstrate a microfluidic flow cytometer cell sorter that uses fast scanning laser excitation sources and photomultiplier tubes (PMTs), coupled with real-time image processing, to image and sort cells based on user-defined spatial features. Dozens of features, including label-free characteristics such as size, shape, scattering pattern, morphology, as well as the spatial distribution of fluorescently labeled biomarkers, greatly expand the available information space in cell analysis. The use of a disposable microfluidic cartridge that is compatible with the imaging system contributes to size reduction and lowered risk of cross contamination. The so-called cameraless design that uses PMTs to replace CCD makes the system more compatible with conventional fluorescence-activated cell sorters and user friendly, while enjoying the sensitivity and benefits of PMTs. The cell image features can be directly extracted from the temporal waveforms or from the reconstructed cell images via temporal-spatial transformation.