Omel'ianchuk L V, Semeshin V F, Alekseeva A L, Pal'chikova I G, Zhimulev I F
Tsitologiia. 2010;52(4):349-53.
Quantitative nuclei DNA content measurement based on Feulgen reaction and the analysis of CCD images was tested. The measurements were performed in monochorome CCD option (650 X 514 pixels) with the wavelength 551 nm. The linear dependence of photomatrix elements signals on the falling light was shown with the use of multigraded light absorption filter. The optimal microscope and camera setting and an approach for elimination of the optic blur were found. We have shown that the proper Feulgen fluorescence does not affect our measurements. Densitometric DNA content measurements of blood cells of four vertebrate species (Gallus domesticus, Danio rerio, Homo sapiens and Rana arvalis) showed good consistence to the literature data (http://www.genomesize.com). The precision of our approach is comparable to the other known methods. Current improvement of CCD technical parameters and wide usage of CCD cameras in biological applications gives perspectives for the suggested approach.
对基于福尔根反应的细胞核DNA含量定量测量及电荷耦合器件(CCD)图像分析进行了测试。测量在单色CCD模式(650×514像素)下、波长为551 nm时进行。通过使用多级光吸收滤光片,显示了光电矩阵元件信号与入射光的线性关系。找到了最佳的显微镜和相机设置以及消除光学模糊的方法。我们已经表明,适当的福尔根荧光不影响我们的测量。对四种脊椎动物(家鸡、斑马鱼、人类和林蛙)血细胞的光密度DNA含量测量结果与文献数据(http://www.genomesize.com)显示出良好的一致性。我们方法的精度与其他已知方法相当。当前CCD技术参数的改进以及CCD相机在生物应用中的广泛使用为所建议的方法提供了前景。