Nonlinear Spectral Imaging of DNA Specimens Derived from Tumor Cells Based on Second Harmonic Generation
LUO Dong-mei1, DENG Xiao-yuan1, ZHUO Shuang-mu2, TAN Shu-wen1, ZHUANG Zheng-fei1, JIN Ying1*
1. College of Biophotonics of South China Normal University, Guangzhou 510631, China 2. Key Laboratory of Optoelectronic Science and Technology for Medicine of Fujian Normal University, Fuzhou 350007, China
Abstract：Second harmonic generation (SHG) is a second-order nonlinear optical process that has symmetry constraints confining signal to regions lacking a center of symmetry. Using SHG microscopy, a variety of tissue structures have noninvasively been imaged by virtue of intrinsic signal generated by structured proteins such as collagen fibrils in connective tissues or the actomyosin lattice of muscle cells. In biochemistry and structure biology, the high-level structures of DNA and protein macro-molecules are similar in constructing mechanism, although DNAs consist of deoxynucleotides and proteins of amino acid residues. The principal purpose of present work is to detect the SHG signal from different DNA samples by spectral imaging technology based on two-photon excited fluorescence (TPEF) and SHG. These DNA samples include the solution of genomic DNA and extracted nuclei, and cultured living cells. Results show that we can obtain the SHG signal from solution of genomic DNA and extracted nuclei in routine condition, but nothing from cultured cell nuclei. After adding a little of absolute ethanol (less than 5% by volume) in culture medium, the SHG signal is detectable in the interested region of nuclei. The findings suggest that the interaction between ethanol and DNA in living cell gives rise to the shift of molecular conformation, and this shift changes some nonlinear optical properties of DNA molecules.
罗冬梅1，邓小元1，卓双木2，谭淑雯1，庄正飞1，金 鹰1* . 肿瘤细胞DNA样品基于二次谐波发生(SHG)的非线性光谱学成像[J]. 光谱学与光谱分析, 2012, 32(03): 800-804.
LUO Dong-mei1, DENG Xiao-yuan1, ZHUO Shuang-mu2, TAN Shu-wen1, ZHUANG Zheng-fei1, JIN Ying1* . Nonlinear Spectral Imaging of DNA Specimens Derived from Tumor Cells Based on Second Harmonic Generation . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2012, 32(03): 800-804.
[1] Denk W, Stickler H, Webb W W. J. Science, 1990, 248(4951): 73. [2] Gannaway J N, Sheppard C J R. J. Opt. Quantum Electron, 1978, 10: 435. [3] Campagnola P J, W ei M, L ewis A, et al. Biophysical Journal, 1999, 77: 3341. [4] Campagnola P J, Clark H A, Mohler W A, et al. J. Biomed. Opt., 2001, 6: 227. [5] Campagnola P J, Millard A C, Terasaki M, et al. Biophysical Journal, 2002, 81(1): 493. [6] Freund I, Deutsch M, Sprecher A J. Biophysical Journal, 1986, 50(4): 693. [7] Meng Han, Lander Zickler, Guenter Giese, et al. Journal of Biomedical Optics, 2004, 9(4): 760. [8] Yi J, Ivan V Tomov, Yim in W, et al. J. Applied Physics Letters, 2005, 86: 133901. [9] Take shi Yasui, Yoshiyuki Tohno, Tsutomu Araki. Journal of Biomedical Optics, 2004, 9(2): 259. [10] Gauderon R, Lukins P B, Sheppard C J R. J. Micron, 2001, 32(7): 685. [11] Zhuang Z F, Liu H P, Guo Z Y, et al. J. Chin. Phys. B, 2010, 19(4): 4262. [12] YAN Dan, WANG Zhu-yuan, CUI Yi-ping(严 丹，王著元，崔一平). J. Optoelectronic Technology(光电子技术学报), 2008，28(2): 129. [13] QU Jun-le, CHEN Dan-ni, YANG Jian-jun, et al(屈军乐，陈丹妮，杨建军, 等). Polytechnic J. of Shenzhen University(深圳大学学报·理工版), 2006, 23(1): 1. [14] Luo T S, Chen J X, Zhuo S M, et al. J. Frontiers of Optoelectronics in China, 2008, (1): 33. [15] Ramanujam N. Neoplasia, 2000, 2: 89. [16] Martine P F, Catherine V B, Eric T, et al. Tetrahedron, 1996, 52: 13569. [17] PENG Xiao-bin, LIANG Shi-qiang(彭小彬，梁世强). J. Chemical Progress (化学进展), 2001, 13(5): 360. [18] Stoller P, Kim B M, Rubenchik A M, et al. J. of Biomedical Optics, 2002 7: 205. [19] Brahms J, Mommaerts W F H. J. Mol.Biol. 1964, 10:73. [20] Ivanov V I, Minchenkova L E, Minyat E E, et al. J. Mol. Biol., 1974, 87: 817. [21] Minchenkova L E, Schyolkina A K, Chernov B K, et al. J. Biomol. Struct. Dyn., 1986, 4: 463. [22] Saenger W. Principles of Nucleic Acid Structure(Springer, New York), 1984. 385. [23] Seeman N C, Rosenberg J M, Rich, A. Proc. Natl. Acad. Sci. USA, 1976, 73: 804. [24] Berg O G, Von Hippel P H. J. Mol. Biol., 1987，193：723.
