Photosynthesis Research

, 102:143 | Cite as

Fluorescence lifetime-resolved imaging

  • Yi-Chun Chen
  • Robert M. Clegg


This is a short account of fluorescence lifetime-resolved imaging, in order to acquaint readers who are not experts with the basic methods for measuring lifetime-resolved signals throughout an image. We present the early FLI (fluorescence lifetime imaging) history, review shortly the instrumentation and experimental design, discuss briefly the fundamentals of the measured fluorescence response, and introduce the basic measurement methodologies. We also emphasize the complex nature of the fluorescence response in FLI signals, and introduce certain analysis methods that are appropriate and informative for complex fluorescence decays. The advantages of model independent analyses are discussed and examples given.


Fluorescence lifetime-resolved imaging Microscopy Model independent analysis Fluorescence lifetime-resolved imaging microscopy Fluorescence lifetime-resolved imaging Spectral FLIM Time-domain Frequency domain 



Fluorescence lifetime-resolved imaging


Fluorescence lifetime-resolved imaging microscopy



We thank Bryan Q. Spring for helping with the Spectral-FLI setup and Shizue Matsubara for introducing us to avocado leaves, and for a wonderful ongoing collaboration and many discussions. We thank Govindjee for many lively and educational discussions about photosynthesis, and for suggestions on the manuscript. 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 B.V. 2009

Authors and Affiliations

  1. 1.Bioengineering DepartmentUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  2. 2.Center for Biophysics and Computational BiologyUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  3. 3.Department of PhysicsUniversity of Illinois at Urbana-Champaign, Loomis Laboratory of PhysicsUrbanaUSA

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