Deciphering fluorescence signals by quantifying separately the excitation intensity from the number of emitters

[摘要]：Conventional fluorescence detection is sensitive to an intricate product of the number of fluorescent emitters times the local excitation intensity. Here, we describe a method to locally quantify the excitation intensity and the number of emitters separately, enabling a clear distinction between the phenomena responsible for a given fluorescence signal. Our technique is based on harmonic excitation modulation and higher-order fluorescence demodulation. It is compatible with a wide range of observations, opening new characterization routes for fluorescence detection or imaging on complex systems, such as plasmonic substrates or scattering media. (C) 2011 Optical Society of America

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