Optimizing Laguerre expansion based deconvolution methods for analysing bi-exponential fluorescence lifetime images

Yongliang Zhang, Yu Chen, David Day-Uei Li

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Abstract

Fast deconvolution is an essential step to calibrate instrument responses in big fluorescence lifetime imaging microscopy (FLIM) image analysis. This paper examined a computationally effective least squares deconvolution method based on Laguerre expansion (LSD-LE), recently developed for clinical diagnosis applications, and proposed new criteria for selecting Laguerre basis functions (LBFs) without considering the mutual orthonormalities between LBFs. Compared with the previously reported LSD-LE, the improved LSD-LE allows to use a higher laser repetition rate, reducing the acquisition time per measurement. Moreover, we extended it, for the first time, to analyze bi-exponential fluorescence decays for more general FLIM-FRET applications. The proposed method was tested on both synthesized bi-exponential and realistic FLIM data for studying the endocytosis of gold nanorods in Hek293 cells. Compared with the previously reported constrained LSD-LE, it shows promising results.
Original languageEnglish
Pages (from-to)13894-13905
Number of pages12
JournalOptics Express
Volume24
Issue number13
DOIs
Publication statusPublished - 14 Jun 2016

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Keywords

  • photon counting
  • image analysis
  • microscopy
  • lifetime-based sensing
  • time-resolved imaging

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