Thunnissen F B, Perdaen H, Forrest J
Department of Pathology, University of Limburg, Maastricht, The Netherlands.
Cytometry. 1992;13(5):485-9. doi: 10.1002/cyto.990130506.
The purpose of this study was to investigate the influence of different cell extraction procedures on relative nuclear DNA content (IOD), nuclear area, and texture features of Feulgen-stained nuclei. In imprints and smears of fine-needle aspirates and suspensions from one human liver specimen, 50 diploid, 50 tetraploid, and 25 octaploid nuclei were measured from each slide. In addition, for DNA measurements, the progressive mean of IOD and tetraploid/diploid and octaploid/diploid ratios was calculated. The results show that the progressive mean of the IOD is constant after measuring 25-30 nuclei. For the three types of specimens, the IOD of diploid nuclei varied slightly. The average coefficient of variation was about 5% for the fine-needle aspirates, imprints, and suspensions. For all tissue sampling methods, the 99% confidence limits of the tetraploid/diploid ratio and octaploid/diploid ratio were within the range of 1.9-2.1 and 3.7-4.3, respectively. The nuclear area and most of the texture features showed a significant difference (p less than 0.01) between the three sampling methods in all nuclear populations. In conclusion, different tissue sampling methods have little or no effect on DNA-related IOD measurements, whereas the outcome of nuclear area and texture features is very dependent of the cell extraction procedure.
本研究的目的是调查不同细胞提取程序对福尔根染色细胞核的相对核DNA含量（IOD）、核面积和纹理特征的影响。在取自一份人类肝脏标本的细针穿刺抽吸物和悬浮液的印片及涂片上，从每张载玻片上测量50个二倍体、50个四倍体和25个八倍体细胞核。此外，为了进行DNA测量，计算了IOD的渐进平均值以及四倍体/二倍体和八倍体/二倍体比率。结果表明，在测量25 - 30个细胞核后，IOD的渐进平均值是恒定的。对于这三种类型的标本，二倍体细胞核的IOD略有变化。细针穿刺抽吸物、印片和悬浮液的平均变异系数约为5%。对于所有组织采样方法，四倍体/二倍体比率和八倍体/二倍体比率的99%置信限分别在1.9 - 2.1和3.7 - 4.3范围内。在所有核群体中，三种采样方法之间的核面积和大多数纹理特征显示出显著差异（p小于0.01）。总之，不同的组织采样方法对与DNA相关的IOD测量几乎没有影响，而核面积和纹理特征的结果非常依赖于细胞提取程序。
