title: Cytometric sorting based on the fluorescence lifetime of spectrally overlapping signals.
abstract: 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.
authors.affiliation: Department of Chemical Engineering, New Mexico State University, Las Cruces, NM 88003-001, USA.
authors.forename: Ruofan
authors.initials: R
authors.lastname: Cao
authors.forename: Varayini
authors.initials: V
authors.lastname: Pankayatselvan
authors.forename: Jessica P
authors.initials: JP
authors.lastname: Houston
mesh_terms: D002469:Cell Separation / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005434:Flow Cytometry / Q000295:instrumentation*; D007089:Image Enhancement / Q000295:instrumentation* / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*