Fluorescence lifetime imaging for the two-photon microscope: time-domain and frequency-domain methods
نویسندگان
چکیده
منابع مشابه
Fluorescence lifetime standards for time and frequency domain fluorescence spectroscopy.
A series of fluorophores with single-exponential fluorescence decays in liquid solution at 20 degrees C were measured independently by nine laboratories using single-photon timing and multifrequency phase and modulation fluorometry instruments with lasers as excitation source. The dyes that can serve as fluorescence lifetime standards for time-domain and frequency-domain measurements are all co...
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We compare frequency-and time-domain formulations of deep-tissue fluorescence imaging of turbid media. Simulations are used to show that time-domain fluorescence tomography, implemented via the asymptotic lifetime-based approach, offers a significantly better separability of multiple lifetime targets than a frequency-domain approach. We also demonstrate experimentally, using complex-shaped phan...
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Two-photon microscopy is a powerful tool for imaging of cells or tissues. However, it presents the drawback of being a laser-scanning technique that involves a long acquisition time for fluorescence-lifetime imaging. Thus it is commonly limited to intensity images that give only indications of the location of fluorophores but do not identify the physicochemical properties and interactions betwe...
متن کاملImproved sensitivity for two-photon frequency-domain lifetime measurement.
We demonstrate a method to improve the measurement sensitivity of two-photon frequency-domain lifetime measurements in poor signal to background conditions. This technique uses sinusoidal modulation of the two-photon excitation source and detection of the second harmonic of the modulation frequency that appears in the emission. Additionally, we present the mathematical model which describes how...
متن کاملTheoretical investigation of the photon efficiency in frequency-domain fluorescence lifetime imaging microscopy.
We investigate the photon efficiency of frequency-domain fluorescence lifetime imaging microscopy, using both theoretical and Monte Carlo methods. Our analysis differs from previous work in that it incorporates the data fitting process used in real experiments, allows for the arbitrary choice of excitation and gain waveforms, and calculates lifetimes as well as associated F-values from higher h...
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ژورنال
عنوان ژورنال: Journal of Biomedical Optics
سال: 2003
ISSN: 1083-3668
DOI: 10.1117/1.1586704