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In fluorescence imaging, both fluorescence yield and lifetime are of great importance. Traditionally, with the frequency-domain data, two parameters can be directly recovered through a nonlinear formulation. However, the reconstruction accuracy highly depends on initial guesses. To overcome this hurdle, we propose the linear scheme via an inverse complex-source formulation. Using the real and imaginary parts of the frequency-domain data, the proposed method is fully linear; it is not sensitive to initial guesses and is stable with high-level noise. Meanwhile, the algorithm is efficient, and the reconstruction takes one or a few iterations. In addition, the colocalization constraint due to the unique feature of fluorescence imaging is imposed to enhance algorithm performance. The algorithms are tested with simulated data.

Fully linear reconstruction method for fluorescence yield and lifetime through inverse complex-source formulation: simulation studies