Sensitivity evaluation of NBD-SCN towards cysteine/homocysteine and its bioimaging applications.
A push-pull fluorogenic reagent, NBD-SCN, was applied for specific detection of cysteine (Cys) and homocysteine (Hcy). Replacing thiocyanato group with Cys/Hcy increased the push-pull characteristic of the probe and resulted in emission of fluorescence. The fluorescent response of the probe toward Cys/Hcy was significantly higher than toward glutathione and other amino acids. The probe showed a 470- and 745-fold fluorescence enhancement at 550 nm and detection limit of 2.99 and 1.43 nM for Cys and Hcy, respectively. Time-dependent fluorescence assays showed that the fluorescence intensity reached a plateau within 20s after addition of Cys and within 10 min after addition of Hcy. Furthermore, the fluorescence images of Cys/Hcy in Raw 264.7 cells were obtained after adding this probe to the cells. These results indicate that NBD-SCN not only possesses good selectivity and sensitivity for Cys/Hcy but also can penetrate cells for Cys/Hcy bioimaging.
Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, 75 Bo-Ai Street, Hsinchu 300, Taiwan.
Yen-Hao
YH
Chen
Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, 75 Bo-Ai Street, Hsinchu 300, Taiwan.
Jia-Chun
JC
Tsai
Department of Biological Science and Technology, Chung Hwa University of Medical Technology, 89, Wunhua 1st Street, Rende, Tainan 717, Taiwan.
Tsan-Hwang
TH
Cheng
Translational Research Center, Cancer Center, and Department of Obstetrics and Gynecology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
Shyng-Shiou
SS
Yuan
Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, 75 Bo-Ai Street, Hsinchu 300, Taiwan; Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 807, Taiwan. Electronic address: ymwang@mail.nctu.edu.tw.
Yun-Ming
YM
Wang
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods; D002460:Cell Line; D003545:Cysteine / Q000032:analysis*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006710:Homocysteine / Q000032:analysis*; D057230:Limit of Detection; D008264:Macrophages / Q000737:chemistry / Q000166:cytology; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D061848:Optical Imaging / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods
D005456:Fluorescent Dyes; D006710:Homocysteine; D003545:Cysteine
Cysteine; Fluorescent probe; Homocysteine; ICT; NBD