High Resolution Fluorescence Lifetime Maps from Minimal Photon Counts

Mohamadreza Fazel, Sina Jazani, Lorenzo Scipioni, Alexander Vallmitjana, Enrico Gratton, Michelle A. Digman, Steve Pressé

Research output: Contribution to journalArticlepeer-review

Abstract

Fluorescence lifetime imaging microscopy (FLIM) may reveal subcellular spatial lifetime maps of key molecular species. Yet, such a quantitative picture of life necessarily demands high photon budgets at every pixel under the current analysis paradigm, thereby increasing acquisition time and photodamage to the sample. Motivated by recent developments in computational statistics, we provide a direct means to update our knowledge of the lifetime maps of species of different lifetimes from direct photon arrivals, while accounting for experimental features such as arbitrary forms of the instrument response function (IRF) and exploiting information from empty laser pulses not resulting in photon detection. Our ability to construct lifetime maps holds for arbitrary lifetimes, from short lifetimes (comparable to the IRF) to lifetimes exceeding interpulse times. As our method is highly data efficient, for the same amount of data normally used to determine lifetimes and photon ratios, working within the Bayesian paradigm, we report direct blind unmixing of lifetimes with subnanosecond resolution and subpixel spatial resolution using standard raster scan FLIM images. We demonstrate our method using a wide range of simulated and experimental data.

Original languageEnglish (US)
Pages (from-to)1015-1025
Number of pages11
JournalACS Photonics
Volume9
Issue number3
DOIs
StatePublished - Mar 16 2022

Keywords

  • Bayesian
  • FLIM
  • Gaussian processes
  • confocal
  • lifetime
  • resolution

ASJC Scopus subject areas

  • Biotechnology
  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Electrical and Electronic Engineering

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