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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editor: Gregory W. Faris
  • Vol. 4, Iss. 8 — Jul. 30, 2009

Light Tolerance of R-Phycoerythrin and a Tandem Conjugate Observed by Single Molecule Recrossing Events

Sean M. Burrows, Payal Patel, and Dimitri Pappas

Applied Spectroscopy, Vol. 63, Issue 6, pp. 709-715 (2009)


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Abstract

The recrossing of single molecules in a probe volume was used to investigate light harvesting and energy transfer between R-phycoerythrin (R-PE) and a tandem conjugate dye. The normalized recrossing ratio, Nr/Nt, was defined as the number of molecules that reenter the probe volume (Nr) to the total number of molecules detected (Nt). The energy transfer process in phycobiliproteins was studied as a function of excitation irradiation and irradiation time. This was achieved by investigating the average baseline-subtracted fluorescence intensity, normalized molecular recrossing ratio (Nr/Nt), and the number of molecules detected per second. The photon saturation irradiance of the R-PE and the tandem conjugate were compared with each other, showing that energy transfer to the tandem dye significantly improves photostability and light tolerance of the phycobiliprotein. The Nr/Nt ratio was used to study the photophysical properties of R-phycoerythrin and the tandem conjugate Streptavidin R-Phycoerythrin-AlexaFluor-647 (PE-647). Normalized molecular recrossings showed that energy transfer to a tandem conjugate could reduce the formation of triplet states in R-phycoerythrin and extend the light tolerance of certain phycobiliproteins.

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Vol. 4, Iss. 8 Virtual Journal for Biomedical Optics

Citation
Sean M. Burrows, Payal Patel, and Dimitri Pappas, "Light Tolerance of R-Phycoerythrin and a Tandem Conjugate Observed by Single Molecule Recrossing Events," Appl. Spectrosc. 63, 709-715 (2009)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=as-63-6-709

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