JBO Letters

Label-free in vivo imaging of human leukocytes using two-photon excited endogenous fluorescence

[+] Author Affiliations
Yan Zeng, Qiqi Sun, Seng Khoon Teh, Wei Zhang

Hong Kong University of Science and Technology, Department of Electronic and Computer Engineering, Biophotonics Research Laboratory, Clear Water Bay, Kowloon, Hong Kong SAR, China

Bo Yan

Hong Kong University of Science and Technology, Division of Life Science, State Key Laboratory of Molecular Neuroscience, Clear Water Bay, Kowloon, Hong Kong SAR, China

Zilong Wen

Hong Kong University of Science and Technology, Division of Life Science, State Key Laboratory of Molecular Neuroscience, Clear Water Bay, Kowloon, Hong Kong SAR, China

Hong Kong University of Science and Technology, School of Science and Institute for Advanced Study, Center of Systems Biology and Human Health, Clear Water Bay, Kowloon, Hong Kong SAR, China

Jianan Y. Qu

Hong Kong University of Science and Technology, Department of Electronic and Computer Engineering, Biophotonics Research Laboratory, Clear Water Bay, Kowloon, Hong Kong SAR, China

Hong Kong University of Science and Technology, School of Science and Institute for Advanced Study, Center of Systems Biology and Human Health, Clear Water Bay, Kowloon, Hong Kong SAR, China

J. Biomed. Opt. 18(4), 040504 (Apr 02, 2013). doi:10.1117/1.JBO.18.4.040504
History: Received October 25, 2012; Revised February 27, 2013; Accepted March 8, 2013
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Abstract

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.

Figures in this Article
© 2013 Society of Photo-Optical Instrumentation Engineers

Citation

Yan Zeng ; Bo Yan ; Qiqi Sun ; Seng Khoon Teh ; Wei Zhang, et al.
"Label-free in vivo imaging of human leukocytes using two-photon excited endogenous fluorescence", J. Biomed. Opt. 18(4), 040504 (Apr 02, 2013). ; http://dx.doi.org/10.1117/1.JBO.18.4.040504


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