Cao Ruofan, Pankayatselvan Varayini, Houston Jessica P
Department of Chemical Engineering, New Mexico State University, Las Cruces, NM 88003-001, USA.
Opt Express. 2013 Jun 17;21(12):14816-31. doi: 10.1364/OE.21.014816.
Flow cytometry is a well-established and powerful high- throughput fluorescence measurement tool that also allows for the sorting and enrichment of subpopulations of cells expressing unique fluorescence signatures. Owing to the reliance on intensity-only signals, flow cytometry sorters cannot easily discriminate between fluorophores that spectrally overlap. In this paper we demonstrate a new method of cell sorting using a fluorescence lifetime-dependent methodology. This approach, referred to herein as phase-filtered cell sorting (PFCS), permits sorting based on the average fluorescence lifetime of a fluorophore by separating fluorescence signals from species that emit differing average fluorescence lifetimes. Using lifetime-dependent hardware, cells and microspheres labeled with fluorophores were sorted with purities up to 90%. PFCS is a practical approach for separating populations of cells that are stained with spectrally overlapping fluorophores or that have interfering autofluorescence signals.
流式细胞术是一种成熟且强大的高通量荧光测量工具，它还能够对表达独特荧光特征的细胞亚群进行分选和富集。由于仅依赖强度信号，流式细胞分选仪难以轻易区分光谱重叠的荧光团。在本文中，我们展示了一种使用基于荧光寿命的方法进行细胞分选的新方法。这种方法在此称为相位过滤细胞分选（PFCS），它通过分离来自发射不同平均荧光寿命的物质的荧光信号，允许基于荧光团的平均荧光寿命进行分选。使用基于寿命的硬件，对标记有荧光团的细胞和微球进行分选，纯度高达90%。PFCS是一种实用的方法，可用于分离用光谱重叠荧光团染色或具有干扰性自发荧光信号的细胞群体。
