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Fluorescence Lifetime Imaging Comes of Age How to Do It and How to Interpret It

  • Yi-Chun Chen
  • Bryan Q. Spring
  • Robert M. CleggEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 875)

Abstract

Fluorescence lifetime imaging (FLI) has been used widely for measuring biomedical samples. Practical guidelines on taking successful FLI data are provided to avoid common errors that arise during the measurement. Several methods for analyzing and interpreting FLI results are also introduced; e.g., a model-free data analysis method called the polar plot allows visualization and analysis of FLI data without iterative fitting, and an image denoising algorithm called variance-stabilizing-transform TI Haar helps to elucidate the information of a complex biomedical sample. The instrument considerations and data analysis of Spectral-FLI are also discussed.

Key words

FLIM FLI Fluorescence lifetime imaging microscopy Polar plot Chebyshev transform TI Haar denoising Spectral FLIM 

Notes

Acknowledgement

Yi-Chun Chen thanks the Taiwan Merit Scholarships (TMS-094-1-A-036) for financial support.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Yi-Chun Chen
    • 1
  • Bryan Q. Spring
    • 2
  • Robert M. Clegg
    • 3
    • 1
    • 2
    Email author
  1. 1.Bioengineering DepartmentUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  2. 2.Center for Biophysics and Computational BiologyUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  3. 3.Loomis Laboratory of Physics, Department of PhysicsUniversity of Illinois at Urbana-ChampaignUrbanaUSA

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