当前位置: X-MOL 学术ACS Nano › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Nanoscale Properties of Human Telomeres Measured with a Dual Purpose X-ray Fluorescence and Super Resolution Microscopy Gold Nanoparticle Probe
ACS Nano ( IF 17.1 ) Pub Date : 2017-11-07 00:00:00 , DOI: 10.1021/acsnano.7b07064
J. Charles G. Jeynes 1 , Kalotina Geraki 2 , Christopher Jeynes 3 , Mi Zhaohong 4 , Andrew A. Bettiol 4 , Eva Latorre 5 , Lorna Wendy Harries 5 , Christian Soeller 6
Affiliation  

Techniques to analyze human telomeres are imperative in studying the molecular mechanism of aging and related diseases. Two important aspects of telomeres are their length in DNA base pairs (bps) and their biophysical nanometer dimensions. However, there are currently no techniques that can simultaneously measure these quantities in individual cell nuclei. Here, we develop and evaluate a telomere “dual” gold nanoparticle-fluorescent probe simultaneously compatible with both X-ray fluorescence (XRF) and super resolution microscopy. We used silver enhancement to independently visualize the spatial locations of gold nanoparticles inside the nuclei, comparing to a standard QFISH (quantitative fluorescence in situ hybridization) probe, and showed good specificity at ∼90%. For sensitivity, we calculated telomere length based on a DNA/gold binding ratio using XRF and compared to quantitative polymerase chain reaction (qPCR) measurements. The sensitivity was low (∼10%), probably because of steric interference prohibiting the relatively large 10 nm gold nanoparticles access to DNA space. We then measured the biophysical characteristics of individual telomeres using super resolution microscopy. Telomeres that have an average length of ∼10 kbps, have diameters ranging between ∼60–300 nm. Further, we treated cells with a telomere-shortening drug and showed there was a small but significant difference in telomere diameter in drug-treated vs control cells. We discuss our results in relation to the current debate surrounding telomere compaction.

中文翻译:

双重目的X射线荧光和超分辨率显微镜金纳米粒子探针测量的人类端粒的纳米级性能。

在研究衰老和相关疾病的分子机制时,分析人类端粒的技术势在必行。端粒的两个重要方面是它们在DNA碱基对中的长度(bps)及其生物物理纳米尺寸。但是,目前没有可以同时测量单个细胞核中这些数量的技术。在这里,我们开发并评估了一种与X射线荧光(XRF)和超分辨率显微镜同时兼容的端粒“双”金纳米粒子荧光探针。与标准QFISH(原位定量荧光)相比,我们使用银增强来独立可视化核内金纳米颗粒的空间位置杂交)探针,在〜90%处显示出良好的特异性。为了提高灵敏度,我们使用XRF基于DNA /金结合比计算了端粒长度,并与定量聚合酶链反应(qPCR)测量结果进行了比较。灵敏度低(约10%),可能是由于空间干扰阻止了相对较大的10 nm金纳米颗粒进入DNA空间。然后,我们使用超分辨率显微镜测量了单个端粒的生物物理特性。端粒的平均长度约为10 kbps,直径范围约为60-300 nm。此外,我们用端粒缩短的药物处理细胞,结果显示在药物处理的细胞与对照细胞中,端粒直径存在微小但显着的差异。我们讨论与当前有关端粒紧缩的辩论有关的结果。
更新日期:2017-11-08
down
wechat
bug