Immunogold labeling of chromosomes for scanning electron microscopy: A closer look at phosphorylated histone H3 in mitotic metaphase chromosomes of Hordeum vulgare
Immunogold labeling of chromosomes for scanning electron microscopy: A closer look at phosphorylated histone H3 in mitotic metaphase chromosomes of Hordeum vulgare
复制标题
用于扫描电子显微镜的染色体免疫金标记：仔细观察大麦有丝分裂中期染色体中的磷酸化组蛋白 H3
作者：
E. Schroeder;Andreas Houben;Gerhard Wanner
采用纳米金间接免疫金标记法，对大麦有丝分裂染色体中丝氨酸10位的组蛋白H3进行了高分辨率的扫描电子显微镜检测。该方法允许在三维环境中对信号进行定位和量化。由于染色体结构保存完好，目前在纳米几十年的范围内的结合部位(染色单体、平行基质纤维、螺线管)的表征是可能的。利用立体分析可以对标签进行量化和三维定位。该方法的局限性涉及到保存染色体超微结构、免疫反应物进入固定染色质的可及性和非特异性标记方面的挑战。讨论了银和金增强之间的区别和标记效率的现状。
High-resolution detection of phosphorylated histone H3 at serine 10 in mitotic barley chromosomes for scanning electron microscopy was shown using a novel application of indirect immunogold labeling with Nanogold. This method permits localization and quantification of signals in a three-dimensional context. Because the chromosome structure is well preserved, characterization of binding sites (chromomeres, parallel matrix fibers, solenoids), currently in the realm of nanometer decades, is possible. Quantification and three-dimensional localization of labels is possible with stereoscopic analysis. Limitations of the method pertain to the challenges in preservation of chromosome ultrastructure, accessibility of immunoreactants into the fixed chromatin and unspecific labeling. The differences between silver and gold enhancement and the current status of labeling efficiency are addressed.
影响因子：
4
作者：
E. Kaszás;W. Cande
通讯作者：
E. Kaszás;W. Cande
组蛋白 H3 变体 CENP-A 的着丝粒形成染色质规范。
影响因子：
4
作者：
VanHooser,AA;Ouspenski,II;Gregson,HC;Starr,DA;Yen,TJ;Goldberg,ML;Yokomori,K;Earnshaw,WC;Sullivan,KF;Brinkley,BR
通讯作者：
Brinkley,BR