Stepanov S I, Konyshev V N, Kotlovanova L V, Roganov A P
Molecular and Radiation Biophysics Division, Petersburg Nuclear Physics Institute, Leningrad, Russia.
Cytometry. 1996 Apr 1;23(4):279-83. doi: 10.1002/(SICI)1097-0320(19960401)23:4<279::AID-CYTO3>3.0.CO;2-C.
As the study of metaphase chromosomes with flow cytometry presupposes mixing all chromosomes from many cells before analysis, important information is lost. To overcome this limitation we have developed a novel cell-oriented flow cytometric method for chromosome analysis. A flow cytometer supplied with a special device for disruption of metaphase chromosomes is the heart of the method. Cells with stained chromosomes are pressed into the disruption device and then converted in batches of chromosomes. Chromosome fluorescence and time intervals between fluorescence pulses are registered. There are large time intervals between neighboring batches because a sparse cell suspension is used and, in turn, there are small time intervals inside batches. Taking account of time intervals, it is possible to identify individual cell flow karyotypes. This highly productive method for individual cell analysis (100-200 cells/min) ensures detecting the changes in cell flow karyotypes, i.e., under- and overrepresentation of chromosomes, aberrations, and amplification of DNA.
由于利用流式细胞术对中期染色体进行研究需要在分析前将来自许多细胞的所有染色体混合在一起，重要信息因此丢失。为克服这一局限，我们开发了一种用于染色体分析的全新的面向细胞的流式细胞术方法。配备有用于破坏中期染色体的特殊装置的流式细胞仪是该方法的核心。带有染色染色体的细胞被压入破坏装置，然后被转化为成批的染色体。记录染色体荧光以及荧光脉冲之间的时间间隔。相邻批次之间存在较大的时间间隔，这是因为使用的是稀疏的细胞悬液，相应地，批次内部的时间间隔较小。考虑到时间间隔，就有可能识别单个细胞的流式核型。这种用于单个细胞分析的高效方法（每分钟100 - 200个细胞）能够确保检测到细胞流式核型的变化，即染色体的数量不足和过多、畸变以及DNA扩增。
