Imaging Endogenous Bilirubins with Two-Photon Fluorescence of Bilirubin Dimers

Yu Fang Shen, Ming Rung Tsai, Shao Chieh Chen, Yun Shiuan Leung, Chien Tai Hsieh, Yu Shing Chen, Fu Lien Huang, Rofeamor P. Obena, Medel Manuel L. Zwulueta, Hsin Yi Huang, Wen Jeng Lee, Kuo Chun Tang, Chun Ta Kung, Min Huey Chen, Dar Bin Shieh, Yu Ju Chen, Tzu Ming Liu, Pi Tai Chou, Chi Kuang Sun

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Abstract

(Graph Presented) On the basis of an infrared femtosecond Cr:forsterite laser, we developed a semiquantitative method to analyze the microscopic distribution of bilirubins. Using 1230 nm femtosecond pulses, we selectively excited the two-photon red fluorescence of bilirubin dimers around 660 nm. Autofluorescences from other endogenous fluorophores were greatly suppressed. Using this distinct fluorescence measure, we found that poorly differentiated hepatocellular carcinoma (HCC) tissues on average showed 3.7 times lower concentration of bilirubins than the corresponding nontumor parts. The corresponding fluorescence lifetime measurements indicated that HCC tissues exhibited a longer lifetime (500 ps) than that of nontumor parts (300 ps). Similarly, oral cancer cell lines had longer lifetimes (>330 ps) than those of nontumor ones (250 ps). We anticipate the developed methods of bilirubin molecular imaging to be useful in diagnosing cancers or studying the dynamics of bilirubin metabolisms in live cells.

Original languageEnglish
Pages (from-to)7575-7582
Number of pages8
JournalAnalytical chemistry
Volume87
Issue number15
DOIs
Publication statusPublished - 2015 Aug 4

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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    Shen, Y. F., Tsai, M. R., Chen, S. C., Leung, Y. S., Hsieh, C. T., Chen, Y. S., Huang, F. L., Obena, R. P., Zwulueta, M. M. L., Huang, H. Y., Lee, W. J., Tang, K. C., Kung, C. T., Chen, M. H., Shieh, D. B., Chen, Y. J., Liu, T. M., Chou, P. T., & Sun, C. K. (2015). Imaging Endogenous Bilirubins with Two-Photon Fluorescence of Bilirubin Dimers. Analytical chemistry, 87(15), 7575-7582. https://doi.org/10.1021/acs.analchem.5b01903