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

Abstract

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.

Fig. 1

(Color online) Reconstruction results for case 1 (no background fluorescence).

Fig. 2

(Color online) Reconstruction results for case 2 (contrast = 2) with correct background as the initial guess.

Fig. 3

(Color online) Reconstruction results for case 3 (contrast = 2) with zero background as the initial guess.

Fig. 4

(Color online) Reconstruction results for case 4 (contrast = 2) with 5% noise, correct background, and zero background as the initial guess in methods 1 and 2, respectively.

Fig. 5

(Color online) Reconstruction results for case 5 (contrast = 2) with 10% noise, correct background, and zero background as the initial guess in methods 1 and 2, respectively.