Research Papers

Study of cadmium-induced cytotoxicity using two-photon excitation endogenous fluorescence microscopy

[+] Author Affiliations
Dong Li

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

Mildred S. Yang, Tao Lin

Hong Kong Baptist University, Department of Biology, Kowloon, Hong Kong, China

Wei Zheng, Jianan Y. Qu

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

J. Biomed. Opt. 14(5), 054028 (October 23, 2009). doi:10.1117/1.3250293
History: Received March 05, 2009; Revised August 06, 2009; Accepted August 11, 2009; Published October 23, 2009
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We demonstrate that using time-resolved two-photon excitation endogenous fluorescence microscopy, the cadmium (Cd)-induced cellular toxic level can be assessed by the free-to protein-bound reduced nicotinamide adenine dinucleotide (free/bound NADH) ratio in a living cell. NADH fluorescence excited at 730nm is captured at different times following exposure to cadmium at a variety of concentrations. The temporal characteristics of NADH fluorescence from mitochondrial and nuclear compartments are analyzed, respectively. The results show that cadmium induces a significant increase of the free/bound NADH ratio in mitochondria and nucleus, caused by the inhibition effect on the electron transport chain (ETC) and the stimulating effect on the glycolysis pathway, respectively. It is found that induction of metallothionein (MT) in cells occurs after 4h of exposure to a sublethal concentration of Cd and reaches a peak at 6h. More importantly, the increase in MT level can effectively suppress the elevation of the free/bound NADH ratio caused by a subsequent exposure to a higher concentration of Cd, indicating that MT plays a key role in protecting cells from Cd-induced toxicity. Our findings show that the free/bound NADH ratio can potentially be used as a sensitive indicator of toxic and carcinogenic actions induced by Cd.

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© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Dong Li ; Mildred S. Yang ; Tao Lin ; Wei Zheng and Jianan Y. Qu
"Study of cadmium-induced cytotoxicity using two-photon excitation endogenous fluorescence microscopy", J. Biomed. Opt. 14(5), 054028 (October 23, 2009). ; http://dx.doi.org/10.1117/1.3250293


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