Use of Combined Scanning Electrochemical and Fluorescence Microscopy for Detection of Reactive Oxygen Species in Prostate Cancer Cells
- 26 September 2013
- journal article
- letter
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 85 (20), 9417-9421
- https://doi.org/10.1021/ac402367f
Abstract
Release of ROS from prostate cancer (PC3) cells was studied using scanning electrochemical microscopy (SECM) and fluorescence microscopy. One-directional lateral scan SECM was used as a rapid and reproducible tool for simultaneous mapping of cell topography and reactive oxygen species (ROS) release. Fluorescence microscopy was used in tandem to monitor the tip position, in addition to providing information on intracellular ROS content via the use of ROS-reactive fluorescent dyes. A unique tip current (iT) vs lateral distance profile was observed when the tip potential (ET) was set at -0.65 V. This profile reflects the combined effects of topographical change and ROS release at the PC3 cell surfaces. Differentiation between topographical-related and ROS-induced current change was achieved by comparing the scans collected at -0.65 and -0.85 V. The effects of other parameters such as tip to cell distance, solvent oxygen content, and scan direction on the profile of the scan were systematically evaluated. Cells treated with tert-butyl hydroperoxide, a known ROS stimulus, were also evaluated using the lateral scanning approach. Overall, the SECM results correlate well with the fluorescence results. The extracellular ROS level detected at the SECM tip was found to be similar to the intracellular ROS level monitored using fluorescence microscopy. While the concentration of each contributing ROS species has not been determined and is thus part of the future study, here we have successfully demonstrated the use of a simple two-potential lateral scan approach for analysis of ROS released by living cells under real physiological conditions.Keywords
This publication has 32 references indexed in Scilit:
- Assessment of multidrug resistance on cell coculture patterns using scanning electrochemical microscopyProceedings of the National Academy of Sciences of the United States of America, 2013
- Cisplatin effects on evolution of reactive oxygen species from single human bladder cancer cells investigated by scanning electrochemical microscopyJournal of Inorganic Biochemistry, 2012
- In Vivo Imaging of Ca2+, pH, and Reactive Oxygen Species Using Fluorescent Probes in PlantsAnnual Review of Plant Biology, 2011
- Real-time mapping of a hydrogen peroxide concentration profile across a polymicrobial bacterial biofilm using scanning electrochemical microscopyProceedings of the National Academy of Sciences of the United States of America, 2011
- Triton X-100 concentration effects on membrane permeability of a single HeLa cell by scanning electrochemical microscopy (SECM)Proceedings of the National Academy of Sciences of the United States of America, 2010
- Elevated Cytosolic Na + Increases Mitochondrial Formation of Reactive Oxygen Species in Failing Cardiac MyocytesCirculation, 2010
- Fluorescence Lifetime Measurements and Biological ImagingChemical Reviews, 2010
- Association of reactive oxygen species levels and radioresistance in cancer stem cellsNature, 2009
- Oxidative Stress Is Inherent in Prostate Cancer Cells and Is Required for Aggressive PhenotypeCancer Research, 2008
- Real-time monitoring of reactive oxygen species production during differentiation of human monocytic cell lines (THP-1)Analytica Chimica Acta, 2005