Quantitative estimation of the total sulfide concentration in live tissues by two-photon microscopy

Chang Su Lim, Sajal Kumar Das, Sun Young Yang, Eun-Sun Kim, Hoon-Jai Chun, Bong-Rae Cho

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20 Citations (Scopus)


Hydrogen sulfide (H2S) is a newly recognized transmitter, which protects various organs from oxidative stress. In this article, we report a ratiometric two-photon probe, TFCA, which can be excited by 750 nm femtosecond pulses, shows a 110-fold increase in the intensity ratio upon reaction with HS- and high selectivity for HS- and can visualize the total sulfide ([H2S] + [HS-]) distribution in live tissue by two-photon microscopy (TPM). We also developed a kinetic method to quantitatively estimate the total sulfide concentration ([H2S] + [HS-]) in live tissues. The kinetic method allowed us to measure the observed rate constants (kobs) for the sulfide-induced deazidation reaction of TFCA in live cells and tissues using TPM. The total sulfide concentration was calculated by using kobs = k2[HS -], with the k2 value determined in HEPES/EtOH (1/1, pH = 7.2), and [H2S]/[HS-] = [H+]/Ka. The total sulfide concentration was found to be nearly zero in HeLa cells and 4-7 μM in rat colon tissues.

Original languageEnglish
Pages (from-to)9288-9295
Number of pages8
JournalAnalytical Chemistry
Issue number19
Publication statusPublished - 2013 Oct 1


ASJC Scopus subject areas

  • Analytical Chemistry

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