Abstract

The deconvolution process is a key step for quantitative evaluation of fluorescence lifetime imaging microscopy (FLIM) samples. By this process, the fluorescence impulse responses (FluoIRs) of the sample are decoupled from the instrument response (InstR). In blind deconvolution estimation (BDE), the FluoIRs and InstR are jointly extracted from a dataset with minimal a priori information. In this work, two BDE algorithms are introduced based on linear combinations of multi-exponential functions to model each FluoIR in the sample. For both schemes, the InstR is assumed with a free-form and a sparse structure. The local perspective of the BDE methodology assumes that the characteristic parameters of the exponential functions (time constants and scaling coefficients) are estimated based on a single spatial point of the dataset. On the other hand, the same exponential functions are used in the whole dataset in the global perspective, and just the scaling coefficients are updated for each spatial point. A least squares formulation is considered for both BDE algorithms. To overcome the nonlinear interaction in the decision variables, an alternating least squares (ALS) methodology iteratively solves both estimation problems based on non-negative and constrained optimizations. The validation stage considered first synthetic datasets at different noise types and levels, and a comparison with the standard deconvolution techniques with a multi-exponential model for FLIM measurements, as well as, with two BDE methodologies in the state of the art: Laguerre basis, and exponentials library. For the experimental evaluation, fluorescent dyes and oral tissue samples were considered. Our results show that local and global perspectives are consistent with the standard deconvolution techniques, and they reached the fastest convergence responses among the BDE algorithms with the best compromise in FluoIRs and InstR estimation errors.

Highlights

  • Fluorescence microscopy has become a powerful tool to characterize the chemical properties of tissue samples [1,2,3,4]

  • We introduced two new blind deconvolution estimation (BDE) algorithms based on a linear combination of multiexponential functions for the fluorescence impulse responses (FluoIRs) modeling

  • For the global perspective, the exponential functions are assumed common to all the points in the dataset, and just their scaling coefficients are updated for each spatial point

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Summary

RESEARCH ARTICLE

Blind deconvolution estimation by multiexponential models and alternated least squares approximations: Free-form and sparse approach.

OPEN ACCESS
Introduction
Blind deconvolution estimation
HLðτkÞ eÀ
Estimation of FluoIR parameters
Estimation of InstR parameters
Initial conditions
Synthetic and experimental validation
Synthetic evaluation
Synthetic Dye
Experimental evaluation with oral tissue samples
Squamous cell carcinoma Squamous cell carcinoma Dysplasia Benign Benign
Conclusions
Findings
Author Contributions

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