Esa A, Trakhtenbrot L, Hausmann M, Rauch J, Brok-Simoni F, Rechavi G, Ben-Bassat I, Cremer C
Institute of Applied Physics, Heidelberg, F.R. Germany.
Anal Cell Pathol. 1998;16(4):211-22. doi: 10.1155/1998/764986.
A new fluorescence in situ hybridization (FISH) technique called Fast-FISH in combination with semi-automated image analysis was applied to detect numerical aberrations of chromosomes 8 and 12 in interphase nuclei of peripheral blood lymphocytes and bone marrow cells from patients with acute myelogenous leukemia (AML) and chronic lymphocytic leukemia (CLL). Commercially available alpha-satellite DNA probes specific for the centromere regions of chromosome 8 and chromosome 12, respectively, were used. After application of the Fast-FISH protocol, and microscopic images of the fluorescence-labelled cell nuclei were recorded by the true color CCD camera Kappa CF 15 MC and evaluated quantitatively by computer analysis on a PC. These results were compared to results obtained from the same type of specimens using the same analysis system but with a standard FISH protocol. In addition, automated spot counting after both FISH techniques was compared to visual spot counting after standard FISH. A total number of about 3,000 cell nuclei was evaluated. For quantitative brightness parameters, a good correlation between standard FISH labelling and Fast-FISH was found. Automated spot counting after Fast-FISH coincided within a few percent to automated and visual spot counting after standard FISH. The examples shown indicate the reliability and reproducibility of Fast-FISH and its potential for automatized interphase cell diagnostics of numerical chromosome aberrations. Since the Fast-FISH technique requires a hybridization time as low as 1/20 of established standard FISH techniques, omitting most of the time consuming working steps in the protocol, it may contribute considerably to clinical diagnostics. This may especially be interesting in cases where an accurate result is required within a few hours.
一种名为快速荧光原位杂交（Fast-FISH）的新荧光原位杂交技术与半自动图像分析相结合，用于检测急性髓性白血病（AML）和慢性淋巴细胞白血病（CLL）患者外周血淋巴细胞和骨髓细胞间期核中8号和12号染色体的数目畸变。分别使用了针对8号染色体和12号染色体着丝粒区域的市售α-卫星DNA探针。应用Fast-FISH方案后，用真彩色CCD相机Kappa CF 15 MC记录荧光标记细胞核的显微图像，并在个人电脑上通过计算机分析进行定量评估。将这些结果与使用相同分析系统但采用标准FISH方案从相同类型标本中获得的结果进行比较。此外，还将两种FISH技术后的自动斑点计数与标准FISH后的目视斑点计数进行了比较。共评估了约3000个细胞核。对于定量亮度参数，发现标准FISH标记与Fast-FISH之间具有良好的相关性。Fast-FISH后的自动斑点计数与标准FISH后的自动和目视斑点计数相差仅几个百分点。所示实例表明了Fast-FISH的可靠性和可重复性及其在染色体数目畸变的自动化间期细胞诊断中的潜力。由于Fast-FISH技术所需的杂交时间仅为既定标准FISH技术的1/20，省略了方案中大部分耗时的工作步骤，因此它可能对临床诊断有很大贡献。在需要在几小时内获得准确结果的情况下，这可能特别有意义。
