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

作　　者：张良玉[1] 邓新宇[1] 傅松滨[1] 

机构地区：[1]哈尔滨医科大学遗传教研室,150086

出　　处：《中国地方病学杂志》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)

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.

关 键 词：原子力显微镜 染色体 双微体 AFM DMS 分辨率 

分 类 号：Q-336[生物学]