Bazett-Jones D P, Hendzel M J, Kruhlak M J
Department of Cell Biology and Anatomy, University of Calgary, Alberta, Canada.
Micron. 1999 Apr;30(2):151-7. doi: 10.1016/s0968-4328(99)00019-0.
We describe a method to image selectively the protein-based architecture in the eukaryotic cell nucleus using nitrogen and phosphorus mapping. In addition, we describe a method to determine total mass as well as stoichiometric relationships between protein and RNA. This method is illustrated using particulate structures in the nucleus called interchromatin granules. In so doing, we demonstrate that these granules contain heterogeneous nuclear RNA, and have an average protein and RNA content of 3.094 and 1.672 MDa, respectively. We also tested the sensitivity of phosphorus detection by exogenously applying purified duplex DNA to the surfaces of thin sections, and have shown that structures as small as single molecules of duplex DNA can be detected in situ using these electron spectroscopic imaging techniques.
我们描述了一种利用氮和磷映射选择性成像真核细胞核中基于蛋白质的结构的方法。此外，我们还描述了一种确定总质量以及蛋白质与RNA之间化学计量关系的方法。使用细胞核中称为染色质间颗粒的颗粒结构对该方法进行了说明。通过这样做，我们证明这些颗粒含有异质核RNA，并且平均蛋白质和RNA含量分别为3.094 MDa和1.672 MDa。我们还通过将纯化的双链DNA外源应用于薄片表面来测试磷检测的灵敏度，并且已经表明使用这些电子光谱成像技术可以原位检测到小至双链DNA单分子的结构。
