技术论文 “Sorting of Chromosomes on FACSAriaTM SORP for the Preparation of Painting Probes“于2016年8月25日发表于Cytometry Part A。并且，同期的“Commentary”栏目刊发了澳大利亚Walter and Eliza Hall Institute 的Simon Monard的评论文章“Chromosome Sorting for the Masses?” 对黄粤组技术成果做出了高度评价。

    高纯度的染色体可以通过MoFlo MLS和BD influx分选获得，这些都是配备有高功率的水冷激光器的空气激发型细胞分选仪。FACSAria 配备了低功率的激光器，光胶耦合石英流动池以及光路收集系统。但是，是否高纯度的小鼠和人的染色体通过BD FACSAriaTM Special Order Research Product (FACSAria SORP)分析获得还未知。这里，我们通过FACSAria SORP （设置50 mW 355 nm激光器； 20 mW 444 nm激光器）获得了高分辨率的小鼠淋巴细胞核和正常男性外周血单核细胞的流式核型。而且利用荧光原位杂交（FISH）证明了来源于分选染色体制备的染色体涂染探针的高纯度和信号特异性。值得注意的是，人的12号染色体在流式核型中从9-12染色体簇分离开来，并且通过12号染色体三体的人ES细胞系B2-C7和B2-B8的FISH进行了证实。总的来说，我们的发现证实了利用FACSAria SORP能够获得高分辨率的流式核型。而且，通过FISH分析证实了分选染色体的高纯度。

    黄粤课题组的贾玉艳助理研究员为该研究论文的第一作者，课题组的博士研究生程丽，刘光助理研究员和张美丽助理研究员也参与该研究；黄粤研究员为本文通讯作者。该研究由国家自然科学基金资助，并且申请了一项发明专利。

 文章网址：http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.22934/abstract

【摘要】High purity chromosome sorting can be performed on instruments such as MoFlo MLS and BD influx, which are stream-in-air sorters equipped with water-cooled high power lasers. The FACSAria is a true fixed alignment, low laser powered instrument with a quartz flow cell gel-coupled to the collection optics. However, whether high purity mouse and human chromosomes can be obtained by sorting on the BD FACSAriaTM Special Order Research Product (FACSAria SORP) remains to be determined. Here, we report that the high resolution flow karyotype of mouse lymphocytes and normal male human peripheral blood mononuclear cells (hPBMCs) can be obtained on the FACSAria SORP using laser power settings of 50 mW for 355 nm and 20 mW for 444 nm excitation. Furthermore, the use of Fluorescence in situ hybridization (FISH) confirmed that chromosome paints prepared from the sorted chromosomes demonstrated high purity and signal specificity. Notably, human chromosome 12 was separated from the chromosome 9–12 cluster in the flow karyotype, and its identity was confirmed using FISH in trisomy 12 human ES cell lines B2-C7 and B2-B8. In addition, multicolor FISH (mFISH) with human chromosome painting probes to 13,18, 21, and sex chromosomes X and Y showed high signal specificity in hPBMCs. Taken together, our findings demonstrated that high resolution flow karyotype can be obtained using FACSAria SORP. Moreover, a FISH analysis confirmed high purity of the sorted chromosomes. Additionally, in contrast to centromeric satellite probes, chromosome painting probes with high specificity are more suitable for detection of chromosome aberrations, such as deletions and translocations, in prenatal diagnosis. © 2016 International Society for Advancement of Cytometry

Figure 3. Open in figure viewerDownload Powerpoint slide Verification of chromosome sorting purity. FISH with whole-chromosome paint probes (WCP) in mouse lymphocytes (A) or hPBMCs (B). Mouse chromosome 12 could be distinguished from chromosome 14 by FISH according to the centromere feature. Scale bar: 10 μm. The images are representative of three independent experiments.