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原子力显微镜在双微体研究中的方法学探讨

Method discussion of double minute chromosomes studied by atomic forc e microscopy
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摘要 目的探讨影响原子力显微镜(atomic force microscopy,AFM)观察双微体(double minute chromosomes,DMs)的各种因素,包括原子力显微镜成像模式的选择,以及样品制备过程中各种条件的优化。方法培养耐氨甲喋呤(MTX)300μmol/L的小鼠成纤维细胞系3T3R500,收获中期染色体后制片,胃蛋白酶消化标本,梯度酒精脱水,最后用原子力显微镜观察染色体中期分裂相。结果优化条件后,在原子力显微镜下很容易找到分裂相。图像扫描显示,染色体在分散程度和清晰度上都有很大提高,双微体的成像质量也有极大的改善。结论优化染色体制片条件及选择恰当的原子力显微镜成像模式可获得高质量的DMs图片。 Objective To discuss the influencing factors during observing doubl e minute chromosomes(DMs) by atomic force microscopy(AFM),such as choosing prope r image mode and optimizing conditions in the procedure of preparing sample for AFM. Methods After mouse fibroblast cell line 3T3R500MTX300 was cultured,meta phasic chromosomes were obtained into specimen,followed by digestion by pepsin, dehydration by 75% ethanol, 8% ethanol and 100% ethanol and observation under AF M. Results It was very easy to find chromosomes under AFM after conditions ha d been optimized with improved scattering and clear chromosome images. Conclusio ns High-quality images of double minutes can be obtained under optimized cond itions and in proper image modes of AFM.
出处 《中国地方病学杂志》 CAS CSCD 北大核心 2005年第3期323-325,共3页 Chinese Jouranl of Endemiology
基金 国家自然科学基金资助项目(30370783)
关键词 原子力显微镜 染色体 双微体 AFM DMS 分辨率 Atomic force microscopy Chromosome Double minute chromosomes
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参考文献13

  • 1曾洁铭,曾耀英,蔡继业,肇静娴.原子力显微镜在生命科学中的应用[J].中国病理生理杂志,2001,17(2):180-185. 被引量:10
  • 2张汝芝,朱文元,汪晨.三氧化二砷诱导Cloundman S91恶性黑素瘤细胞凋亡的研究[J].临床皮肤科杂志,2003,32(3):121-123. 被引量:6
  • 3陈勇,蔡继业,朱大明,洪岸.PC12细胞凋亡及其细胞膜表面超微结构的原子力显微镜形态学观察[J].电子显微学报,2003,22(4):311-314. 被引量:4
  • 4邓新宇,张钰,曾力,国立秋,闫永达,董申,张贵寅,李璞,傅松滨.原子力显微镜在双微体形态学研究中的应用[J].Acta Genetica Sinica,2003,30(9):881-885. 被引量:2
  • 5Pauletti G, Lai E, Attardi G. Early appearance and long-term persistence of the submicroscopic extrachromosomal elements (amplisomes) containing the amplified DHFR genes in human cell lines [J ]. PNAS, 1990,87: 2955-2959.
  • 6Hamlin JL, Leu TH, Vaughn JP, et al. Amplification of DNA sequences in mammalian cells[J]. Prog Nucleic Acid Res Mol Biol,1991,41:203-239.
  • 7Santos NC, Castanho MA. An overview of the biophysical applications of atomic force microscopy [J]. Biophys Chem,2004,107(2): 133-149.
  • 8Obataya I, Nakamura C, Han S,et al. Mechanical sensing of the penetration of various nanoneedles into a living cell using atomic force microscopy [ J ]. Biosens Bioelectron, 2005,20 (8): 1652-1655.
  • 9Berdyyeva T, Woodworth CD, Sokolov I. Visualization of cytoskeletal elements by the atomic force microscope[J]. Ultramicroscopy,2005,102(3): 189-198.
  • 10Avci R, Schweitzer M, Boyd RD,et al. Comparison of antibodyantigen interactions on collagen measured by conventional immunological techniques and atomic force microscopy[J]. Langmuir,2004,20(25 ): 11053-11063.

二级参考文献21

  • 1Hamkalo B A, Peggy J Farnham, Randal Johnston, Robert Schimke.Ultrastructure features of minute chromosomes in a methotrexate-resistant mouse 3T3 cell line. Cell Biology, 1985,82(4), 1026 - 1030.
  • 2Teru Kanda, Kevin F Sullivan, Geoffrey M Wahl. Histone-GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells. Current Biology, 1998,8 : 377 - 385.
  • 3T J McMaster, M 0 Winfield, A A Baker, A Karp, M J Miles. Chromosome classification by atomic force microscopy volume measurement. J Vac Sci Technol, 1996,14(2) : 1438 - 1442.
  • 4Mario B Viani, Lia I Pietrsanta, James B Thompson, Ami Chand, Ilse C Gebeshuber, Johannes H Kindt, Michael Richter, Helen G Hansma,Paul K Hansma. Probing protein-protein interactions in real time. Nature Structural Biology ,2000,7(8) :644 - 647.
  • 5Alastair Smith. Atomic force microscopy. Microbiology Today, 26(99) :54 - 55.
  • 6Thalhammer S, Stark R W, Muller S, Wienberg J, Heckl W M. The atomic force microscope as a new mierodissecting tool for the generation of genetic probes. J Struct Biol, 1997,119(2) :232 - 237.
  • 7Iwabuchii S, Moil T, Ogawa K, Sato K, Saito M, Morita Y, Ushiki T,Tamiya E. Atomic force microscope-based dissection of human metaphase chromosomes and high resolutional imaging by carbon nanotube tip. Arch Histol Cytol,2002 ,65(5) :473 - 479.
  • 8Peter J Hahn, Barbara Nevaldine, John A Longo. Molecular structure and evolution of double-Minute chromosomes in methotrexate-resistant cultured mouse cells. Molecular and Cellilar Biology, 1992, 12(7) :2911 - 2918.
  • 9E M Jack, J J Waters, C J Harrison. A scanning microscopy study of double minutes from a human tumour cell line. Cytogenet Cell Genet,1987,44:49 - 52.
  • 10J B Rattner, C C Lin. Ultrastructureal organization of double minute chromosomes and HSR regions in human colon carcinoma cells. Cytogenet Cell Genet, 1984,38 : 176 - 181.

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