A ratiometric two-photon fluorescent probe for imaging hydrogen sulfide in lysosomes.
Hydrogen sulfide (H2S) is a kind of gaseous signalling molecule that plays pivotal role in various biological processes. So far, it is still a challenge to develop convenient and reliable methods for H2S detection in lysosomes. Herein, we developed a novel ratiometric two-photon fluorescent probe LR-H2S for imaging H2S in lysosomes. Upon the addition of H2S (using Na2S as a donor) to LR-H2S in buffer solution, the azide group is reduced to amino group and subsequently the carbamate ester is cleaved by 1,6-elimination, resulting in a fluorescence emission increase at 541nm and a concomitant emission decrease at 475nm. Under two-photon excitation of 840nm, an 80-fold fluorescence ratio (F541/F475) enhancement was observed with a wide linear range of 25-2500μM. The detection limit was calculated to be 0.70μM based on 3σ/k method, indicating that the probe can detect H2S with a high sensitivity. The probe also shows excellent selectivity toward H2S among other biological interference species and features with low cytotoxicity and favorable two-photon properties. Furthermore, LR-H2S can easily localize in lysosomes and vividly illuminate endogenous/exogenous H2S level and distribution in lysosomes of living SGC-7901 cells.
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China.
Wenqi
W
Feng
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China.
Zhiqiang
Z
Mao
College of Science, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: liulingzhi_1981@mail.hzau.edu.cn.
Lingzhi
L
Liu
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China. Electronic address: zhhliu@whu.edu.cn.
Zhihong
Z
Liu
D045744:Cell Line, Tumor; D002470:Cell Survival; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D057054:Molecular Imaging / Q000379:methods*; D017785:Photons
D005456:Fluorescent Dyes; D006862:Hydrogen Sulfide
Bio-imaging; H(2)S; Lysosome-targeted; Ratiometric; Two-photon