In Situ Sensing of Reactive Oxygen Species on Dye-Stained Single DNA Molecules under Illumination

Langmuir : the ACS Journal of Surfaces and Colloids
Joseph R PyleJixin Chen

Abstract

Reactive oxygen species (ROS) are a necessary evil in many biological systems and have been measured with fluorescent probes at the ensemble levels both in vitro and in vivo. Measuring ROS generated from a single molecule is important for mechanistic studies, yet measuring ROS near a dye-labeled single-molecule under illumination has been challenging. Here, we use CellROX, a group of ROS probes, to sense ROS near dye-stained DNA that has been flow-stretched and immobilized on a surface. ROS is responsible for the photodamage of DNA molecules under this circumstance. In this report, we confirmed the ROS sensing reaction in bulk solutions and optimized the conditions for single-molecule experiments including the selection of substrates, dye concentrations, probes in the CellROX series, excitation lasers, and emission filter-sets. We observed a correlation between ROS and the dye-labeled DNA and localized the ROS-activated CellROX probe molecules at both the ensemble level and the single-molecule level.

References

Aug 23, 1983·Biochimica Et Biophysica Acta·J M Gutteridge, S Wilkins
May 23, 1995·Proceedings of the National Academy of Sciences of the United States of America·W CaiD C Schwartz
Mar 15, 1996·Nucleic Acids Research·B Akerman, E Tuite
Jan 1, 1997·Free Radical Biology & Medicine·W R Markesbery
Jul 8, 1998·Proceedings of the National Academy of Sciences of the United States of America·J JingD C Schwartz
Nov 15, 2000·American Journal of Physiology. Lung Cellular and Molecular Physiology·V J Thannickal, B L Fanburg
Aug 17, 2001·Journal of the American Chemical Society·C KanonyE Tuite
Mar 21, 2003·The American Journal of Cardiology·David HarrisonHelmut Drexler
May 26, 2004·Drug Resistance Updates : Reviews and Commentaries in Antimicrobial and Anticancer Chemotherapy·Helene PelicanoPeng Huang
Aug 19, 2004·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·Markus F Renschler
Feb 5, 2005·Analytical Biochemistry·Matthew M FerrisRichard A Keller
Jan 25, 2006·Chemico-biological Interactions·M ValkoM Mazur
Jun 8, 2006·Journal of the American Chemical Society·Alexandre FürstenbergEric Vauthey
Dec 5, 2006·Proceedings of the National Academy of Sciences of the United States of America·Alexey Sharonov, Robin M Hochstrasser
Apr 25, 2007·Apoptosis : an International Journal on Programmed Cell Death·Martin OttBoris Zhivotovsky
Jun 27, 2007·Journal of Pharmaceutical Sciences·Tomris Ozben
Jul 21, 2007·Chemistry : a European Journal·Alexandre FürstenbergEric Vauthey
May 31, 2008·Diabetes·Josephine M ForbesMark E Cooper
Nov 5, 2008·Cancer Biology & Therapy·Jie Wang, Jing Yi
Jan 31, 2012·Cellular Signalling·Paul D RayYoshiaki Tsuji
Apr 10, 2012·Analytical Biochemistry·Michael A TyconChristopher J Fecko
Nov 23, 2013·Physical Chemistry Chemical Physics : PCCP·Bo ShuangChristy F Landes
May 23, 2014·Current Biology : CB·Michael Schieber, Navdeep S Chandel
Jan 16, 2016·Biochimica Et Biophysica Acta·Panagiotis Lepetsos, Athanasios G Papavassiliou
Nov 29, 2017·Beilstein Journal of Nanotechnology·Joseph R Pyle, Jixin Chen
Oct 24, 2018·Nano Letters·Xiao-Qiang WangXian-Zheng Zhang
Apr 23, 2019·ACS Applied Materials & Interfaces·Juvinch R VicenteJixin Chen

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