Journal of the American Chemical Society
COMMUNICATION
Next, we tested the abilities of SF1 and SF2 to visualize
changes in H2S levels in live-cell imaging mode using confocal
microscopy (Figure 3). HEK293T cells were incubated with
5 μM SF1 or SF2 for 30 min and then treated with a blank control
or 250 μM NaSH for an additional 30 min; this concentration is
well within the range that has been used to elicit physiological
responses (10À600 μM).10,11,15 A patent increase in intracellular
fluorescence intensity was observed in the H2S-treated cells
(Figure 3b,e) relative the to control samples (Figure 3a,d).
Moreover, bright-field images with Hoescht nuclear staining
confirmed the viability of the cells over the course of the
experiments (Figure 3c,f). Interestingly, SF1 gave a higher
turn-on response than SF2 for the detection of H2S in cells
(Figure S4), which may be due to increased lipophilicity and
cellular retention of SF1 (log P = 1.98 ( 0.1) relative to SF2 (log
P = 1.21 ( 0.15).
To close, we have presented a new approach to biological H2S
detection through the synthesis and evaluation of the azide-based
fluorescent probes SF1 and SF2. These reagents are highly
selective for detection of H2S in aqueous media and can be used
for imaging H2S in living cells. We are actively pursuing more
sensitive and responsive analogues for fluorescence imaging of
H2S in living cells, tissues, and animals as well as the utilization of
these probes to study the endogenous production of H2S in living
cells and its contributions to physiological and pathological
processes.
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’ ASSOCIATED CONTENT
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Supporting Information. Synthesis and experimental
b
details, including procedures for the synthesis and characteriza-
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’ AUTHOR INFORMATION
Corresponding Author
(31) Lippert, A. R.; Keshari, K. R.; Kurhanewicz, J.; Chang, C. J. J.
Am. Chem. Soc. 2011, 133, 3776–3779.
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’ ACKNOWLEDGMENT
This work was supported by Amgen, Astra Zeneca, Novartis,
and the Packard Foundation. E.J.N. acknowledges the Royal
Commission for the Exhibition of 1851 for additional support. C.
J.C. is an Investigator of the Howard Hughes Medical Institute.
We thank Ann Fischer and Michelle Yasukawa (UCB Tissue
Culture Facility) for expert technical assistance.
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