Two-photon fluorescence imaging of intracellular hydrogen peroxide with chemoselective fluorescent probes

Hengchang Guo; Hossein Aleyasin; Scott S. Howard; Bryan C. Dickinson; Vivian S. Lin; Renee E. Haskew-Layton; Chris Xu; Yu Chen; Rajiv R. Ratan

doi:10.1117/1.JBO.18.10.106002

1 October 2013 Two-photon fluorescence imaging of intracellular hydrogen peroxide with chemoselective fluorescent probes

Hengchang Guo, Hossein Aleyasin, Scott S. Howard, Bryan C. Dickinson, Vivian S. Lin, Renee E. Haskew-Layton, Chris Xu, Yu Chen, Rajiv R. Ratan

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Journal of Biomedical Optics, Vol. 18, Issue 10, 106002 (October 2013). https://doi.org/10.1117/1.JBO.18.10.106002

Abstract

We present the application of two-photon fluorescence (TPF) imaging to monitor intracellular hydrogen peroxide H₂O₂ production in brain cells. For selective imaging of H₂O₂ over other reactive oxygen species, we employed small-molecule fluorescent probes that utilize a chemoselective boronate deprotection mechanism. Peroxyfluor-6 acetoxymethyl ester detects global cellular H₂O₂ and mitochondria peroxy yellow 1 detects mitochondrial H₂O₂ . Two-photon absorption cross sections for these H₂O₂ probes are measured with a mode-locked Ti:sapphire laser in the wavelength range of 720 to 1040 nm. TPF imaging is demonstrated in the HT22 cell line to monitor both cytoplasmic H₂O₂ and localized H₂O₂ production in mitochondria. Endogenous cytoplasmic H₂O₂ production is detected with TPF imaging in rat astrocytes modified with d-amino acid oxidase. The TPF H₂O₂ imaging demonstrated that these chemoselective probes are powerful tools for the detection of intracellular H₂O₂.

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Hengchang Guo, Hossein Aleyasin, Scott S. Howard, Bryan C. Dickinson, Vivian S. Lin, Renee E. Haskew-Layton, Chris Xu, Yu Chen, and Rajiv R. Ratan "Two-photon fluorescence imaging of intracellular hydrogen peroxide with chemoselective fluorescent probes," Journal of Biomedical Optics 18(10), 106002 (1 October 2013). https://doi.org/10.1117/1.JBO.18.10.106002

Published: 1 October 2013

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