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Nanoscale Properties of Human Telomeres Measured with a Dual Purpose X-ray Fluorescence and Super Resolution Microscopy Gold Nanoparticle Probe

Jeynes, J. Charles G., Geraki, Kalotina, Jeynes, Christopher, Zhaohong, Mi, Bettiol, Andrew A., Latorre, Eva, Harries, Lorna Wendy and Soeller, Christian (2017) Nanoscale Properties of Human Telomeres Measured with a Dual Purpose X-ray Fluorescence and Super Resolution Microscopy Gold Nanoparticle Probe ACS Nano, 11 (12). pp. 12632-12640.

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Abstract

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.

Item Type: Article
Divisions : Faculty of Engineering and Physical Sciences > Electronic Engineering
Authors :
NameEmailORCID
Jeynes, J. Charles G.
Geraki, Kalotina
Jeynes, Christopherc.jeynes@surrey.ac.uk
Zhaohong, Mi
Bettiol, Andrew A.
Latorre, Eva
Harries, Lorna Wendy
Soeller, Christian
Date : 1 November 2017
Identification Number : 10.1021/acsnano.7b07064
Copyright Disclaimer : Copyright © 2017 American Chemical Society
Uncontrolled Keywords : dSTORM; Nanoparticles; Super resolution microscopy; Telomeres; X-ray fluorescence
Depositing User : Clive Harris
Date Deposited : 14 Mar 2018 14:20
Last Modified : 27 Mar 2018 07:46
URI: http://epubs.surrey.ac.uk/id/eprint/846025

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