两种少量小鼠外周血间期核FISH技术的比较

Comparison of Two Kinds of Peripheral Blood Interphase Nuclei FISH Technology

目的:探讨低渗法与红细胞裂解法应用于小鼠少量外周血间期核荧光原位杂交(fluorescence in situ hybridization,FISH)检测的优缺点,以便于根据实验条件选取最合适的方法。方法:小鼠剪尾巴法获取外周血,采用低渗法或红细胞裂解法破坏红细胞,涂片前先用免疫组化笔在玻片上划好预计涂片区域,重悬沉淀后稍等片刻再吸中间部分用于涂片,涂完后在相差显微镜下观察密度,行常规FISH,比较两种方法处理后的FISH结果。结果:低渗法和红细胞裂解法处理后的片子背景都较干净,密度适中,信号较强。结论:在外周血间期核荧光原位杂交技术中,低渗法和红细胞裂解法都可以获得较满意的FISH结果。低渗法的特点在于试剂便宜且易于获取,但耗时稍长;而红细胞裂解法的特点在于可节省实验时间,但需专用试剂。

Objective： To compare hypotonic solution and red cell lysis buffer used in fluorescence in situ hybridization technology（FISH）. Methods： Peripheral blood is obtained by cutting the tail of mouse by using hypotonic solution or red blood cell lysis buffer to destroy the red blood cells, immunohistochemical pen was used to stroke the expected zone before smear, waiting for a while after resuspending the precipitation and then the middle portion is sucked to smear, the coating density was observed under the phase contrast microscope, FISH was according to the general methods, then the results of FISH were compared. Results： The background of slide is relatively clean processed by both hypotonic solution and red cell lysis buffer method, the density is appropriate, and the signal is powerful.Conclusions： Both hypotonic solution and red cell lysis buffer method can get satisfactory results of FISH of interphase nuclei in peripheral blood, The characteristics of hypotonic solution method is that the reagent is cheap and easy to obtain, but it will take more time; and the red cell lysis buffer method＇ characteristics is that it can save time, but need special reagents.