Linear and Nonlinear Fluorescence Lifetime Imaging for Biosensing Applications
Abstract
Fluorescence techniques, based on both linear and non-linear excitation, have been widely used in biosensing application. Among fluorescence techniques, Fluorescence Lifetime Imaging (FLIM) has gained increased importance because of their robust and versatile feature. FLIM is a technique to visualize the spatial distribution of picosecond to nanosecond excited state lifetimes within microscopic images. In this work, FLIM technique combined with Time Correlated Single Photon Counting (TCPSC) Technology was used to develop several novel biosensing approaches. Using linear or nonlinear excitation source, TCSPC-FLIM enabled real time monitoring of cellular metabolite profiles, as well as longitudinal observation of post-translational enzyme activity such as kinase phosphorylation and acetyltransferase acetylation. This technique will not only provide high spatiotemporal resolution of sensing output but also for the first time, realize real-time biosensing in live intact single cell within 2 D culture, 3D culture and in-vivo animal model, zebrafish.
Degree
Ph.D.
Advisors
Irudayaraj, Purdue University.
Subject Area
Cellular biology|Biomedical engineering|Biophysics
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