Abstract:Quantum dots have many excellent optical properties such as high quantum yield, long fluorescence lifetime, wide excitation spectrum and narrow emission spectrum, tunable emission wavelength and so on, thus have become a newpopular type of fluorescence probes in these years.Quantum-dot-based DNA nanosensor comprising streptavidin-conjugated quantum dots, capture probes with biotin and reporter probes with Cy5 was designed to detect DNA or RNA segments.Capture probes and reporter probes were connected by the target DNA or RNA segments so that quantum dots and Cy5s could be together and FRET (fluorescence resonance energy transfer) could be detected.In the present work, quantum-dot-based DNA nanosensor was combined with ICCD fluorescence microscopy imaging system through the authors’ experiments.Using the total internal reflection fluorescence (TIRF), FRET between quantum dots and Cy5s was recorded by ICCD showing that segments of single-stranded target DNA with 30-base length were detected in solution using DNA nanosensor.When Cy5-ssDNA-Biotins were added into streptavidin-conjugated quantum dots in solution, by real time recording, the FRET efficiency was found to increase with time, which indicated the process of streptavidin-conjugated quantum dots capturing Cy5-ssDNA-Biotins.It was also observed that streptavidin-conjugated quantum dots and Cy5-ssDNA-Biotins could both enter living Chinese hamster ovary cells and have FRET.The process of streptavidin-conjugated quantum dots capturing Cy5-ssDNA-Biotins was detected in the cells as well and Cy5s were photobleached after a long time of irradiation.It has been proved that detecting DNA or RNA segments in living cells with DNA nanosensor is possible.
邓楚芸,李佳敏,马万云*. DNA纳米传感器荧光成像技术[J]. 光谱学与光谱分析, 2010, 30(01): 220-224.
DENG Chu-yun, LI Jia-min, MA Wan-yun*. DNA Nanosensor Fluorescence Imaging Microscopy. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2010, 30(01): 220-224.
[1] Stryer L, Haugland R.Proc.Natl.Acad.Sci.USA, 1967, 58: 719. [2] Jares-Erijman Elizabeth A, Jovin Thomas M.Nature Biotechnology, 2003, 21(11): 1387. [3] Giepmans Ben N G.Adams Stephen R, Ellisman Mark H et al.Science, 2006, 312: 217. [4] Nienhaus Gerd Ulrich.Macromolecular Bioscience, 2006, 6(11): 907. [5] Clapp Aaron R, Medintz Igor L, Mattoussi Hedi.Chem.Phys.Chem., 2006, 7(1): 47. [6] Piston David W, Kremers Gert-Jan.Trends in Biochemical Sciences, 2007, 32(9): 407. [7] Deniz Aarok A, Mukhopadhyay Samrat, Lemke Edward A.Journal of the Royal Society Interface, 2008, 5(18): 15. [8] Zhang Chunyang, Yeh Hsin-Chin, Kuroki Marcos T, et al.Nature Materials, 2005, 4(11), 826. [9] Allen Michael D, Zhang Tin..Angew.Chem.Int.Ed., 2008, 47(3): 500. [10] Zhang Chunying, Johnson Lawrencew.Angew.Chem.Int.Ed., 2007, 46(19): 3482. [11] Nakane Jonathan, Wiggin Matthew, Marziali Andre.Biophysical Journal, 2004, 87(1): 615. [12] Mattheyses Alexa L, Axelrod Daniel, Axelyod Daniel.Journal of Biomedical Optics, 2006, 11(1): 014006. [13] Zhang Chun-yang, Johnson Lawrence W.J.Am.Chem.Soc., 2006, 128(16): 5324. [14] Suzuki Miho, Husimi Yuzuru, Komatsu Hirokazu, et al.J.Am.Chem.Soc., 2008, 130(17): 5720. [15] Diltemiz S E, Say Ridvan, Buyuktiyaki Sibel, el al.Talanta, 2008, 75(4): 890. [16] Vo-Dinh T, Cullum Brian M, Stokes Darid L.Sensors and Actuators B, Chemical, 2001, 74(1-3): 2. [17] WANG Yan-jun, LI Yong-sheng, YANG Quan-yu, et al(王艳君,李永生,杨全玉,等).Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2009, 29(1): 165. [18] LU Jin-ling, CHU Jun, YANG Jie, et al(陆锦玲,储 军,杨 杰,等).Progress in Biochemistry and Biophysics(生物化学与生物物理进展), 2008, 35(3): 268. [19] LIU Chun-chun, HANG Hai-ying(刘春春,杭海英).Progress in Biochemistry and Biophysics(生物化学与生物物理进展), 2006, 33(3): 292.