Lebo R V, Golbus M S, Cheung M C
Am J Med Genet. 1986 Nov;25(3):519-29. doi: 10.1002/ajmg.1320250314.
Our custom dual laser chromosome sorter has been used to characterize and isolate metaphase human chromosomes rapidly for gene mapping purposes. Herein, we tested how well this system could detect unknown abnormal human chromosome constitutions. These results were compared to those of conventional cytogenetic analyses by banding and photomicrography. The sorter was used to analyze each cell line stained with two different stain pairs: DIPI-chromomycin and Hoechst-chromomycin. In 20 min, two histograms representing 2 X 10(5) chromosomes each were collected for each stain pair. A blind study of 11 samples by flow analysis demonstrated excellent concordance between the abnormal chromosomes detected and the diagnoses of Giemsa-banded karyotypes. Aneuploidy was identified by changes in the number of chromosomes in each histogram peak, while rearrangements such as deletions and translocations caused shifts in the histogram peak positions. The direction and distance of histogram peak shifts are directly related to alterations in chromosome size and banding pattern. We conclude that dual-laser flow analysis may provide a rapid approach to the screening and diagnosis of chromosome abnormalities.
我们定制的双激光染色体分拣仪已被用于快速鉴定和分离中期人类染色体，以进行基因定位。在此，我们测试了该系统检测未知异常人类染色体组成的能力。将这些结果与通过显带和显微摄影进行的传统细胞遗传学分析结果进行了比较。分拣仪用于分析用两种不同染色对染色的每个细胞系：二碘化丙啶-放线菌素和 Hoechst-放线菌素。在20分钟内，为每个染色对收集了两个分别代表2×10⁵条染色体的直方图。通过流式分析对11个样本进行的盲法研究表明，检测到的异常染色体与吉姆萨显带核型诊断之间具有高度一致性。通过每个直方图峰中染色体数量的变化来识别非整倍体，而缺失和易位等重排会导致直方图峰位置的移动。直方图峰移动的方向和距离与染色体大小和带型的改变直接相关。我们得出结论，双激光流式分析可能为染色体异常的筛查和诊断提供一种快速方法。
