Sakamoto M, Pinkel D, Mascio L, Sudar D, Peters D, Kuo W L, Yamakawa K, Nakamura Y, Drabkin H, Jericevic Z
Division of Molecular Cytometry, University of California, San Francisco 94143-0808, USA.
Cytometry. 1995 Jan 1;19(1):60-9. doi: 10.1002/cyto.990190108.
This paper describes an evaluation of a semiautomated, multicolor image-analysis system to map cloned probes along metaphase chromosomes. Mapping with this system consists of fluorescence in situ hybridization (FISH) for probe localization, automatic acquisition of multicolor images showing total chromosomal DNA and probe location(s), and automatic determination of the fractional locations of the probes along the chromosomes relative to the short arm telomere (FLpter). The system was evaluated by mapping ten phage and ten cosmid probes previously mapped to chromosome 3 with other procedures. The standard deviations of FLpter measurements averaged 3.4 Mb and 2.6 Mb for phage and cosmid probes, respectively. With this variation, the order of two probes mapped in separate hybridizations could be determined with 95% confidence when their separation was greater than 2.5 Mb. In all cases, the probe locations and order were consistent with previous mapping data. FLpter values were converted to band locations using measurements of the band locations made using digital imaging microscopy. This proved superior to conversions made using ISCN ideograms.
本文描述了一种半自动多色图像分析系统的评估，该系统用于沿中期染色体定位克隆探针。使用该系统进行定位包括用于探针定位的荧光原位杂交（FISH）、自动采集显示总染色体DNA和探针位置的多色图像，以及自动确定探针沿染色体相对于短臂端粒（FLpter）的相对位置。通过定位十个先前用其他方法定位到3号染色体的噬菌体探针和十个黏粒探针来评估该系统。噬菌体探针和黏粒探针的FLpter测量值的标准差分别平均为3.4 Mb和2.6 Mb。有了这种变化，当两个在单独杂交中定位的探针间距大于2.5 Mb时，就可以95%的置信度确定它们的顺序。在所有情况下，探针的位置和顺序与先前的定位数据一致。使用数字成像显微镜测量带位置，将FLpter值转换为带位置。这被证明优于使用国际人类细胞遗传学命名法（ISCN）图表进行的转换。
