Novel fluorescent probes for highly selective two-photon imaging of mitochondria in living cells.
The synthesis and characterization of a pair of novel pyridine cation derivatives possessing two-photon excitation fluorescence (TPEF) properties and selectively staining mitochondria in living SiHa cells within 30 min, CAI and CAEI, were reported. And the green-emitting CAI displayed a much larger two-photon excitation fluorescence action absorption cross-section (δ × Φ) of 328 g at 860 nm in comparison with commercial MitoTracker Green (MTG) with maximum δ × Φ value of 2.18 g at 850 nm. As is known to all, δ × Φ is a crucial parameter to obtain a high-quality microscopic photo in living cells in two-photon microscopy (TPM). Moreover, the fact that the co-localization coefficient between CAI and conventional MitoTracker Red (MTR) was 0.95 in SiHa cells demonstrated specific staining performance of CAI to mitochondria. As biosensors, both CAI and CAEI possessed a number of beneficial properties such as large δ × Φ and Stokes shifts, good membrane permeability, long retention time, high photostability and excellent counterstain compatibility with different biosensors for instance Hoechst 33342 and D 307, which ranked them as one of the best TPEF mitochondrial probes. Furthermore, related mechanism research suggested that their localization properties were dependent on the mitochondrial membrane potential in living cells. And their remarkable properties can extend the investigation on mitochondria in a biological context.
Center of Bio & Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, PR China.
Fang
F
Miao
Center of Bio & Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, PR China.
Weijia
W
Zhang
School of Information Science and Engineering, Shandong University, Jinan 250100, PR China.
Yuming
Y
Sun
Center of Bio & Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, PR China.
Ruoyao
R
Zhang
Center of Bio & Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, PR China.
Yong
Y
Liu
Center of Bio & Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, PR China.
Fuqiang
F
Guo
School of Information Science and Engineering, Shandong University, Jinan 250100, PR China.
Guofen
G
Song
Center of Bio & Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, PR China.
Minggang
M
Tian
Center of Bio & Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, PR China. Electronic address: yuxq@sdu.edu.cn.
Xiaoqiang
X
Yu
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D003287:Contrast Media / Q000138:chemical synthesis; D005456:Fluorescent Dyes / Q000138:chemical synthesis*; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D008928:Mitochondria / Q000648:ultrastructure*; D011725:Pyridines / Q000737:chemistry*; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D003287:Contrast Media; D005456:Fluorescent Dyes; D011725:Pyridines; C023666:pyridine
Bioimaging; Fluorescent probe; High selectivity; Mitochondria; Two-photon