Abstract

Mounting evidence supports the role of hydrogen peroxide (H2O2) in physiological signaling as well as pathological conditions. However, the subtleties of peroxide-mediated signaling are not well understood, in part because the generation, degradation, and diffusion of H2O2 are highly volatile within different cellular compartments. Therefore, the direct measurement of H2O2 in living specimens is critically important. Fluorescent probes that can detect small changes in H2O2 levels within relevant cellular compartments are important tools to study the spatial dynamics of H2O2. To achieve temporal resolution, the probes must also be photostable enough to allow multiple readings over time without loss of signal. Traditional fluorescent redox sensitive probes that have been commonly used for the detection of H2O2 tend to react with a wide variety of reactive oxygen species (ROS) and often suffer from photostablilty issues. Recently, new classes of H2O2 probes have been designed to detect H2O2 with high selectivity. Advances in H2O2 measurement have enabled biomedical scientists to study H2O2 biology at a level of precision previously unachievable. In addition, new imaging techniques such as two-photon microscopy (TPM) have been employed for H2O2 detection, which permit real-time measurements of H2O2in vivo. This review focuses on recent advances in H2O2 probe development and optical imaging technologies that have been developed for biomedical applications.Electronic supplementary materialThe online version of this article (doi:10.1186/2045-3701-4-64) contains supplementary material, which is available to authorized users.

Highlights

  • The role of Hydrogen peroxide (H2O2) as a second messenger, in regulating fundamental biological processes, has been identified not long ago and is increasingly supported by new data [1,2,3,4,5,6,7]

  • Much of the data implicating H2O2 both pathological and physiological roles in cultured cells and in vivo has been acquired using treatment with exogenous H2O2 [19,20], over expression of hydroperoxide inducing enzymes or H2O2 lysing/scavenging agents [1,21], or tampering with reactive oxygen species (ROS) production cellular machinery such as NADPH oxidase (Nox) expressing cells [22]. These studies were critical in establishing crucial biological roles of H2O2, a key to fully understanding the mechanistic bases of redox biology is measuring the amount of H2O2 generated in specific intracellular compartments

  • We describe and compare various methodologies for detection and imaging of H2O2 production in cells and whole organisms

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Summary

Introduction

The role of H2O2 as a second messenger, in regulating fundamental biological processes, has been identified not long ago and is increasingly supported by new data [1,2,3,4,5,6,7]. Confocal microscopy coupled with chemoselective fluorescent reporters permits the imaging of localized intracellular H2O2 levels. Several novel fluorescent probes capable of detecting H2O2 with high selectivity have been reported, and some of them have been used to monitor intracellular H2O2.

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