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.
['Jenkins|Patrick|P|', 'Naivar|Mark A|MA|', 'Houston|Jessica P|JP|']
D000818:Animals D016466:CHO Cells D003198:Computer Simulation D003412:Cricetulus D004867:Equipment Design D005434:Flow Cytometry D005453:Fluorescence D005583:Fourier Analysis D007834:Lasers D008863:Microspheres D008962:Models, Theoretical D017711:Nonlinear Dynamics D012542:Scattering, Radiation
fluorescence lifetime flow cytometry square wave modulation frequency harmonics frequency-domain phase shift