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Journal of Biomedical Optics • Open Access

Label-free in vivo imaging of human leukocytes using two-photon excited endogenous fluorescence
Author(s): Yan Zeng; Bo Yan; Qiqi Sun; Seng Khoon Teh; Wei Zhang; Zilong Wen; Jianan Y. Qu

Paper Abstract

We demonstrate that two-photon excited endogenous fluorescence enables label-free morphological and functional imaging of various human blood cells. Specifically, we achieved distinctive morphological contrast to visualize morphology of important leukocytes, such as polymorphonuclear structure of granulocyte and mononuclear feature of agranulocyte, through the employment of the reduced nicotinamide adenine dinucleotide (NADH) fluorescence signals. In addition, NADH fluorescence images clearly reveal the morphological transformation process of neutrophils during disease-causing bacterial infection. Our findings also show that time-resolved NADH fluorescence can be potentially used for functional imaging of the phagocytosis of pathogens by leukocytes (neutrophils) in vivo. In particular, we found that free-to-bound NADH ratios measured in infected neutrophils increased significantly, which is consistent with a previous study that the energy consumed in the phagocytosis of neutrophils is mainly generated through the glycolysis pathway that leads to the accumulation of free NADH. Future work will focus on further developing and applying label-free imaging technology to investigate leukocyte-related diseases and disorders.

Paper Details

Date Published: 2 April 2013
PDF: 4 pages
J. Biomed. Opt. 18(4) 040504 doi: 10.1117/1.JBO.18.4.040504
Published in: Journal of Biomedical Optics Volume 18, Issue 4
Show Author Affiliations
Yan Zeng, Hong Kong Univ. of Science and Technology (Hong Kong, China)
Bo Yan, Hong Kong Univ. of Science and Technology (Hong Kong, China)
Qiqi Sun, Hong Kong Univ. of Science and Technology (Hong Kong, China)
Seng Khoon Teh, Hong Kong Univ. of Science and Technology (Hong Kong, China)
Wei Zhang, Hong Kong Univ. of Science and Technology (Hong Kong, China)
Zilong Wen, Hong Kong Univ. of Science and Technology (Hong Kong, China)
Jianan Y. Qu, Hong Kong Univ. of Science and Technology (Hong Kong, China)


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