Sensitive detection of intracellular environment of normal and cancer cells by autofluorescence lifetime imaging

被引:29
|
作者
Awasthi, Kamlesh [1 ]
Moriya, Daiki [2 ]
Nakabayashi, Takakazu [3 ]
Li, Liming [2 ]
Ohta, Nobuhiro [1 ]
机构
[1] Natl Chiao Tung Univ, Dept Appl Chem, 1001 Ta Hsueh Rd, Hsinchu 30010, Taiwan
[2] Chitose Inst Sci & Technol, Dept Bio & Mat Photon, Chitose 0668655, Japan
[3] Tohoku Univ, Grad Sch Pharmaceut Sci, Aoba Ku, Sendai, Miyagi 9808578, Japan
关键词
FLIM; Autofluorescence; NADH; FAD; Normal cell; Cancer cell; FLAVIN ADENINE-DINUCLEOTIDE; FLUORESCENCE LIFETIME; ENDOGENOUS FLUORESCENCE; CELLULAR-METABOLISM; ELECTRON-TRANSFER; DYNAMICS; PH; FLAVOPROTEIN; SPECTROSCOPY; MICROSCOPY;
D O I
10.1016/j.jphotobiol.2016.10.023
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Intracellular fluorescence lifetime images of the endogenous fluorophores of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD), which are well known as autofluorescence chromophores, were obtained from rat normal fibroblast cells (WFB) and H-ras oncogene-transfected cancer cells among WFB (W31). The average lifetime of the NADH and FAD autofluorescence was shorter in cancer cells than in normal cells, indicating that the difference in metabolism between healthy and cancer cells alters the conditions for coenzymes such as NADH and FAD and that the autofluorescence lifetime measurement of NADH and FAD is applicable for the noninvasive diagnosis of cancer cells. The pico- and nano-second time-resolved fluorescence spectra of NADH obtained with different time windows were similar in normal and cancer cells, indicating that every fluorescence decay component gives the same spectrum in both cell types. These results as well as the fluorescence lifetime images of exogenous fluorophores stained with sodium pheophorbide a in normal and cancer cells suggest that the difference in the fluorescence lifetime between normal and cancer cells cannot be attributed to a difference in the intracellular pH or refractive index but to the difference in the bound condition between proteins and NADH or FAD under the different intracellular environments of normal and cancer cells. (C) 2016 Elsevier B.V. All rights reserved.
引用
收藏
页码:256 / 265
页数:10
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