Share Email Print
cover

Proceedings Paper

Biophotonic tool for sensing the dynamics of H2O2 extracellular release in stressed cells
Author(s): G. Suarez; Ch. Santschi; S. Dutta-Gupta; L. Juillerat-Jeanneret; O. J. F. Martin
Format Member Price Non-Member Price
PDF $14.40 $18.00
cover GOOD NEWS! Your organization subscribes to the SPIE Digital Library. You may be able to download this paper for free. Check Access

Paper Abstract

Hydrogen peroxide (H2O2) is known to play a multifaceted role in cell physiology mechanisms involving oxidative stress and intracellular signal transduction. Therefore, the development of analytical tools providing information on the dynamics of H2O2 generation remains of utmost importance to achieve further insight in the complex physiological processes of living cells and their response to environmental stress. In the present work we developed a novel optic biosensor that provides continuous real-time quantification of the dynamics of the hydrogen peroxide release from cells under oxidative stress conditions. The biosensor is based on the ultra-sensitive dark field optical detection of cytochrome c (cyt c) that exhibits a narrow absorption peaks in its reduced state (Fe(II)) at λ = 550 nm. In the presence of H2O2 the ferrous heme group Fe(II) is oxidised into Fe(III) providing the spectroscopic information exploited in this approach. Extremely low limit-of-detection for H2O2 down to the subnanomolar range is achieved by combining scattering substrates (eg. polystyrene beads) able to shelter cyt c and an inverted microscope in dark field configuration. The developed biosensor was able to perform real-time detection of H2O2 extracellular release from human promyelocytic leukemia cells (HL-60) exposed to lipopolysaccaride (LPS) that elicits strong immune-response. This biosensing tool is currently being implemented to the real-time detection of superoxide anion (O2.-) and offers the possibility to extend to further oxidative stress biomarkers such as glutathione. More generally, multianalyte and dynamic informations might bring new insights to understand complex cellular metabolisms involved in oxidative-stress-related diseases and cytotoxic responses.

Paper Details

Date Published: 2 February 2012
PDF: 7 pages
Proc. SPIE 8229, Optical Diagnostics and Sensing XII: Toward Point-of-Care Diagnostics; and Design and Performance Validation of Phantoms Used in Conjunction with Optical Measurement of Tissue IV, 822908 (2 February 2012); doi: 10.1117/12.910318
Show Author Affiliations
G. Suarez, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
Ch. Santschi, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
S. Dutta-Gupta, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
L. Juillerat-Jeanneret, Univ. of Lausanne (Switzerland)
O. J. F. Martin, Ecole Polytechnique Fédérale de Lausanne (Switzerland)


Published in SPIE Proceedings Vol. 8229:
Optical Diagnostics and Sensing XII: Toward Point-of-Care Diagnostics; and Design and Performance Validation of Phantoms Used in Conjunction with Optical Measurement of Tissue IV
Gerard L. Coté; Robert J. Nordstrom, Editor(s)

© SPIE. Terms of Use
Back to Top