Cell sorting using a high throughput fluorescence flow cytometer

Abstract

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 bya 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 asorting 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 characteristicsof 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.

Description

[0001] related application [0002] This application claims priority to Provisional Application No. 62 / 309,806, filed March 17, 2016, and is hereby incorporated by reference in its entirety. [0003] government rights [0004] This invention was supported by the National Science Foundation (Grant No. NSF1447381). The government has certain rights in this invention. Background technique [0005] The present invention relates generally to apparatus and methods for determining the characteristics of particles flowing through a flow cytometer, such as via fluorescence analysis of a sample, and more particularly to sorting particles in a flow cytometer based on, for example, their characteristics (e.g. sorting cells) devices and methods. [0006] Isolation of subpopulations or even single cells from heterogeneous populations has numerous applications in modern biology and biomedicine. Some conventional techniques for isolating cell subpopulations include fluorescence-activated ...

AI Technical Summary

Problems solved by technology

Furthermore, conventional sorting methods based on imaged cells generally cannot be used for high-throughput cell isolation applications due to their high latency in making sorting decisions

Images