A NBD-based simple but effective fluorescent pH probe for imaging of lysosomes in living cells.
NBDlyso with lysosome-locating morpholine moiety has been developed as a high selective and sensitive fluorescent pH probe. This probe can respond to acidic pH (2.0-7.0) in a short time (less than 1 min) and not almost change after continuously illuminated for an extended period by ultraviolet light. The fluorescence intensity of NBDlyso enhanced 100-fold in acidic solution, with very good linear relationship (R(2) = 0.996). The pKa of probe NBDlyso is 4.10. Therefore, NBDlyso was used to detect lysosomal pH changes successfully. Besides, X-ray crystallography was used to verify the structure of NBDlyso, and the recognition mechanism involving photo-induced electron transfer was interpreted theoretically by means of DFT and TDDFT calculations skillfully when NBDlyso comes into play under the acidic condition. This probe showed good ability to sense pH change in living cell image.
Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, PR China; Taishan College, Shandong University, Jinan 250100, PR China.
Xiang-Jian
XJ
Cao
Institute of Developmental Biology, School of Life Science, Shandong University, Jinan 250100, PR China.
Li-Na
LN
Chen
Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, PR China; Taishan College, Shandong University, Jinan 250100, PR China.
Xuan
X
Zhang
Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, PR China.
Jin-Ting
JT
Liu
Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, PR China; Taishan College, Shandong University, Jinan 250100, PR China.
Ming-Yu
MY
Chen
Institute of Developmental Biology, School of Life Science, Shandong University, Jinan 250100, PR China; Taishan College, Shandong University, Jinan 250100, PR China.
Qiu-Rong
QR
Wu
Institute of Developmental Biology, School of Life Science, Shandong University, Jinan 250100, PR China. Electronic address: miaojy@sdu.edu.cn.
Jun-Ying
JY
Miao
Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, PR China. Electronic address: bxzhao@sdu.edu.cn.
Bao-Xiang
BX
Zhao
D018360:Crystallography, X-Ray; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000737:chemistry* / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008958:Models, Molecular; D061848:Optical Imaging / Q000379:methods; D010069:Oxadiazoles / Q000737:chemistry*
C542965:7-nitrobenzoxadiazole; D005456:Fluorescent Dyes; D010069:Oxadiazoles
Cell imaging; Fluorescent probe; Intracellular pH; Lysosomes; NBD