Toward the measurement of multiple fluorescence lifetimes in flow cytometry: maximizing multi-harmonic content from cells and microspheres.
Flow cytometry is a powerful means for in vitro cellular analyses where multi-fluorescence and multi-angle light scattering can indicate unique biochemical or morphological features of single cells. Yet, to date, flow cytometry systems have lacked the ability to capture complex fluorescence dynamics due to the transient nature of flowing cells. In this contribution we introduce a simple approach for measuring multiple fluorescence lifetimes from a single cytometric event. We leverage square wave modulation, Fourier analysis, and high frequency digitization and show the ability to resolve more than one fluorescence lifetime from fluorescently-labelled cells and microspheres. Illustration of a flow cytometer capable of capturing multiple fluorescence lifetime measurements; creating potential for multi-parametric, time-resolved signals to be captured for every color channel.
Department of Chemical & Materials Engineering, New Mexico State University, MSC 3805 P.O. Box 30001, Las Cruces, NM 88003-8001, USA.
Patrick
P
Jenkins
DarklingX, 1199 45th Street, Los Alamos, NM 87544, USA.
Mark A
MA
Naivar
Department of Chemical & Materials Engineering, New Mexico State University, MSC 3805 P.O. Box 30001, Las Cruces, NM 88003-8001, USA.
Jessica P
JP
Houston
D000818:Animals; D016466:CHO Cells / Q000502:physiology; D003198:Computer Simulation; D003412:Cricetulus; D004867:Equipment Design; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005453:Fluorescence; D005583:Fourier Analysis; D007834:Lasers; D008863:Microspheres; D008962:Models, Theoretical; D017711:Nonlinear Dynamics; D012542:Scattering, Radiation
flow cytometry; fluorescence lifetime; frequency harmonics; frequency-domain; phase shift; square wave modulation