Kinetics model for the wavelength-dependence of excited-state dynamics of hetero-FRET sensors

Schwarz, J., Leighton, R., Leopold, H. J., Currie, M., Boersma, A. J., Sheets, E. D. & Heikal, A. A., 2017, Ultrafast Nonlinear Imaging and Spectroscopy V. Liu, Z., Shi, K., Khoo, I. C. & Psaltis, D. (eds.). SPIE, 103800S. (Proceedings of SPIE - The International Society for Optical Engineering; vol. 10380).

Research output: Chapter in Book/Report/Conference proceedingConference contributionAcademicpeer-review

  • Jacob Schwarz
  • Ryan Leighton
  • Hannah J. Leopold
  • Megan Currie
  • Arnold J. Boersma
  • Erin D. Sheets
  • Ahmed A. Heikal

Foerster (or fluorescence) resonance energy transfer (FRET) is a powerful tool for investigating protein-protein interactions, in both living cells and in controlled environments. A typical hetero-FRET pair consists of a donor and acceptor tethered together with a linker. The corresponding energy transfer efficiency of a hetero-FRET pair probe depends upon the donor-acceptor distance, relative dipole orientation, and spectral overlap. Because of the sensitivity of the energy transfer efficiency on the donor-acceptor distance, FRET is often referred to as a "molecular ruler". Time-resolved fluorescence approach for measuring the excited-state lifetime of the donor and acceptor emissions is one of the most reliable approaches for quantitative assessment of the energy transfer efficiency in hetero-FRET pairs. In this contribution, we provide an analytical kinetics model that describes the excited-state depopulation of a FRET probe as a means to predicts the time-resolved fluorescence profile as a function of excitation and detection wavelengths. In addition, we used this developed kinetics model to simulate the time-dependence of the excited-state population of both the donor and acceptor. These results should serve as a guide for our ongoing studies of newly developed hetero-FRET sensors (mCerulean3-linker-mCitrine) that are designed specifically for in vivo studies of macromolecular crowding. The same model is applicable to other FRET pairs with the careful consideration of their steady-state spectroscopy and the experimental design for wavelength- dependence of the fluorescence lifetime measurements.

Original languageEnglish
Title of host publicationUltrafast Nonlinear Imaging and Spectroscopy V
EditorsZhiwen Liu, Kebin Shi, Iam Choon Khoo, Demetri Psaltis
ISBN (Electronic)9781510612174
Publication statusPublished - 2017
EventUltrafast Nonlinear Imaging and Spectroscopy V 2017 - San Diego, United States
Duration: 6-Aug-20177-Aug-2017

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
ISSN (Print)0277-786X
ISSN (Electronic)1996-756X


ConferenceUltrafast Nonlinear Imaging and Spectroscopy V 2017
CountryUnited States
CitySan Diego


Ultrafast Nonlinear Imaging and Spectroscopy V 2017


San Diego, United States

Event: Conference


  • excited-state dynamics, fluorescence lifetime, FRET, kinetics model

ID: 109565220