Detection of H2O2 by fluorescence correlation spectroscopy

Etsuro Ito, Satoshi Watabe, Mika Morikawa, Hiromi Kodama, Ryuichi Okada, Toshiaki Miura

Research output: Chapter in Book/Report/Conference proceedingChapter

9 Citations (Scopus)

Abstract

Fluorescence correlation spectroscopy (FCS) is a technique in which measurement of fluorescence intensity fluctuations is used to clarify dynamic molecular interactions within a very small space in a solution containing a small number of fluorescent molecules. The FCS-based analysis gives the average number and average diffusion time of the fluorescent molecules during their passage through a very small space. One advantage of FCS is that physical separation between free and bound fluorescent probes is not required because the properties of fluorescence fluctuations are accounted for. Therefore, when fluorescent probes are bound with proteins by peroxidase and hydrogen peroxide (H2O2), FCS enables us to detect H2O 2 with high sensitivity. In addition, because H2O 2 is generated by oxidase-catalyzed reactions, a highly sensitive method for detecting H2O2 is applicable to the measurement of low levels of various oxidases and their substrates, such as glucose. We here describe the protocol of a de novo, highly sensitive method for the measurement of H2O2 and glucose using FCS.

Original languageEnglish
Title of host publicationHydrogen Peroxide and Cell Signaling, Part A
PublisherAcademic Press Inc.
Pages135-143
Number of pages9
ISBN (Print)9780124058835
DOIs
Publication statusPublished - 2013
Externally publishedYes

Publication series

NameMethods in Enzymology
Volume526
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Blood
  • Fluorescence correlation spectroscopy
  • Glucose
  • Hydrogen peroxide
  • Tetramethyl rhodamine
  • Tyramide

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Ito, E., Watabe, S., Morikawa, M., Kodama, H., Okada, R., & Miura, T. (2013). Detection of H2O2 by fluorescence correlation spectroscopy. In Hydrogen Peroxide and Cell Signaling, Part A (pp. 135-143). (Methods in Enzymology; Vol. 526). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-405883-5.00008-9