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Cell Sorting Using A High Throughput Fluorescence Flow Cytometer

机译:使用高通量荧光流式细胞仪进行细胞分选

摘要

In one aspect, a method of sorting cells in a flow cytometry system is disclosed, which includes illuminating a cell with radiation having at least two optical frequencies shifted from one another by a radiofrequency to elicit fluorescent radiation from the cell, detecting the fluorescent radiation to generate temporal fluorescence data, and processing the temporal fluorescence data to arrive at a sorting decision regarding the cell without generating an image (i.e., a pixel-by-pixel image) of the cell based on the fluorescence data. In other words, while the fluorescence data can contain image data that would allow generating a pixel-by-pixel fluorescence intensity map, the method arrives at the sorting decision without generating such a map. In some cases, the sorting decision can be made with a latency less than about 100 microseconds. In some embodiments, the above method of sorting cells can have a sub-cellular resolution, e.g., the sorting decision can be based on characteristics of a component of the cell. In some embodiments in which more than two frequency-shifted optical frequencies are employed, a single radiofrequency shift is employed to separate the optical frequencies while in other such embodiments a plurality of different radiofrequency shifts are employed.
机译:在一个方面,公开了一种在流式细胞仪系统中分选细胞的方法,该方法包括用具有至少两个彼此偏移的光学频率的辐射照射辐射的细胞以引起来自细胞的荧光辐射,检测到产生时间荧光数据,并且处理时间荧光数据以得出关于细胞的分类决定,而无需基于荧光数据产生细胞的图像(即,逐像素图像)。换句话说,尽管荧光数据可以包含允许生成逐像素的荧光强度图的图像数据,但是该方法在不生成这种图的情况下达到了分类决定。在某些情况下,可以以小于约100微秒的等待时间做出排序决定。在一些实施例中,上述分类细胞的方法可以具有亚细胞分辨率,例如,分类决策可以基于细胞成分的特征。在采用多于两个频移的光学频率的一些实施例中,采用单个射频移位来分离光学频率,而在其他这样的实施例中,采用多个不同的射频移位。

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