Two-photon excited fluorescence emission from hemoglobin

Qiqi Sun; Yan Zeng; Wei Zhang; Wei Zheng; Yi Luo; Jianan Y. Qu

doi:10.1117/12.2076685

5 March 2015 Two-photon excited fluorescence emission from hemoglobin

Qiqi Sun, Yan Zeng, Wei Zhang, Wei Zheng, Yi Luo, Jianan Y. Qu

Author Affiliations +

Proceedings Volume 9329, Multiphoton Microscopy in the Biomedical Sciences XV; 93290O (2015) https://doi.org/10.1117/12.2076685
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

Hemoglobin, one of the most important proteins in blood, is responsible for oxygen transportation in almost all vertebrates. Recently, we discovered two-photon excited hemoglobin fluorescence and achieved label-free microvascular imaging based on the hemoglobin fluorescence. However, the mechanism of its fluorescence emission still remains unknown. In this work, we studied the two-photon excited fluorescence properties of the hemoglobin subunits, heme/hemin (iron (II)/(III) protoporphyrin IX) and globin. We first studied the properties of heme and the similar spectral and temporal characteristics of heme and hemoglobin fluorescence provide strong evidence that heme is the fluorophore in hemoglobin. Then we studied the fluorescence properties of hemin, globin and methemoglobin, and found that the hemin may have the main effect on the methemoglobin fluorescence and that globin has tryptophan fluorescence like other proteins. Finally, since heme is a centrosymmetric molecule, that the Soret band fluorescence of heme and hemoglobin was not observed in the single photon process in the previous study may be due to the parity selection rule. The discovery of heme two-photon excited fluorescence may open a new window for heme biology research, since heme as a cofactor of hemoprotein has many functions, including chemical catalysis, electron transfer and diatomic gases transportation.

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Qiqi Sun, Yan Zeng, Wei Zhang, Wei Zheng, Yi Luo, and Jianan Y. Qu "Two-photon excited fluorescence emission from hemoglobin", Proc. SPIE 9329, Multiphoton Microscopy in the Biomedical Sciences XV, 93290O (5 March 2015); https://doi.org/10.1117/12.2076685

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KEYWORDS

Luminescence

Proteins

Single photon

Biology

Signal processing

Molecules

Blood

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