Kahn E, Philippe C, Frouin F, Di Paola R, Bernheim A
Institut National de la Santé et de la Recherche Médicale U494, Centre Hospitalier Universitaire Pitié-Salpêtrière, Paris.
Anal Quant Cytol Histol. 1998 Dec;20(6):477-82.
To analyze, with fluorescent probes, by three-dimensional (3D) emission patterns, fluorescence in situ hybridization (FISH) preparations (cosmids, telomeres) and to perform factor analysis of medical image sequences (FAMIS); to use FISH to track relevant DNA sequences in cell nuclei during interphase and in mitotic chromosomes; and to use cytogenetic techniques, resulting in flat preparations of whole cells that are assumed to preserve probe access to their targets.
The study design entailed labeling targets by probes (sequences labeled by fluorescein isothiocyanate) in nuclei and/or chromosomes stained by propidium iodide. Visualization of targets was improved when 3D sequences of images obtained on a single photomultiplier detector of the confocal microscope by z stepping were investigated by FAMIS.
Factors and factor images showed that targets could be detected and differentiated in focal planes inside nuclei or chromosomes.
It is possible to localize cosmids in cell nuclei at interphase and telomeres in mitotic chromosomes by means of 3D sequences of images.
使用荧光探针通过三维（3D）发射模式分析荧光原位杂交（FISH）制剂（黏粒、端粒），并对医学图像序列进行因子分析（FAMIS）；使用FISH追踪间期细胞核和有丝分裂染色体中的相关DNA序列；使用细胞遗传学技术，制作假定能使探针接近其靶标的全细胞平面制剂。
该研究设计包括用探针（异硫氰酸荧光素标记的序列）标记细胞核和/或用碘化丙啶染色的染色体中的靶标。当通过FAMIS研究在共聚焦显微镜的单个光电倍增管探测器上通过z轴步进获得的图像的3D序列时，靶标的可视化得到了改善。
因子和因子图像显示，在细胞核或染色体内部的焦平面中可以检测到并区分靶标。
通过图像的3D序列能够在间期将黏粒定位在细胞核中，并在有丝分裂染色体中定位端粒。
