Supporting Information Copyright Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, 2008
Highly Sensitive and Fast Responsive Fluorescence Turn-on Chemodosimeter for Cu 2+ and its Application in Live Cell Imaging Mengxiao Yu, [a] Mei Shi, [a] Zhigang Chen, [a] Fuyou Li,* [a] Xinxin Li, [b] Yanhong Gao, [c] Jia Xu, [a] Hong Yang, [a] Zhiguo Zhou, [a] Tao Yi, [a] and Chunhui Huang* [a] [a] Department of Chemistry & Laboratory of Advanced Materials Fudan University 220 Handan Road, Shanghai 200433 (PR China) [b] School of Materials Science and Engineering East China University of Science and Technology 130 Meilong Road, Shanghai 200237 (PR China) [c] Hospital of Xinhua Medicine School of Shanghai Jiaotong University 1665 Kongjiang Road, Shanghai 200092 (PR China) [
Part 1. The supplementary materials, including UV-vis and fluorescence spectra, charge numbers of atoms in 1 and 4, ESI mass spectra, XPS spectrum, fluorescence images, and the result of trypan blue viability test.. Part 2. Colour versions of Figures 2, 5, 7 and 8, Scheme 3, and Table 1 in Text
Part 1. The supplementary materials. Figure S1. Absorption spectra of 20 μm 4 upon addition of 10 100 μm Hg 2+ in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution. Inset: Absorbance at 566 nm as a function of Hg 2+ concentration. Figure S2. Fluorescence response of 20 μm 4 upon addition of 10 100 μm Hg 2+ in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution (λ ex = 510 nm). Inset: Emission intensity at 591 nm as a function of Hg 2+ concentration.
Figure S3. Fluorescence response of 20 μm 4 upon addition of 10 100 μm Cu 2+ in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution (λ ex = 510 nm). Inset: Emission intensity at 580 nm as a function of Cu 2+ concentration.
Figure S4. Fluorescence spectra of 20 µm 4 and 100 µm Cu 2+ upon the addition of EDTA in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution (λ ex = 510 nm). The final concentrations of EDTA were 100 µm and 1mM, respectively. Figure S5. Fluorescence spectra of 20 µm 4 and 40 µm Hg 2+ upon the addition of EDTA in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution (λ ex = 510 nm). The final concentrations of EDTA were 100 µm and 1mM, respectively.
Figure S6. The charge numbers of atoms in 4. Figure S7. The charge numbers of atoms in 1.
Figure S8. ESI mass spectrum (positive) of 20 µm 4 in the presence of 2 equiv of Hg 2+ in CH 3 CN/ammonium acetate (0.1 M, 3:7, v/v) solution, indicating the formation of 5. Figure S9. ESI mass spectrum (positive) of 20 µm 4 in the presence of 2 equiv of Cu 2+ in CH 3 CN/ammonium acetate (0.1 M, 3:7, v/v) solution, indicating the formation of 6. Figure S10. ESI mass spectrum (positive) of 20 µm 4 in the presence of 0.5 equiv of Hg 2+ in ethanol, indicating the formation of intermediate carbodiimide 7. The solution was nearly colorless and weakly fluorescent, suggesting the molecules of 7 adopt a spirolactam form.
Figure S11. Fluorescence spectra of 20 µm 4 upon addition of 100 µm Cu + and Cu 2+ in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution (λ ex = 510 nm). Figure S12. XPS spectrum of 4 with 0.5 equiv. of Cu 2+. The data analysis was carried out by using the RBD AugerScan 3.21 software provided by RBD Enterprises. Binding energies were calibrated by using the containment carbon (C1s = 284.6eV).
Figure S13. ESI mass spectrum (positive) of 20 µm 4 in the presence of 0.5 equiv of Cu 2+ in ethanol, indicating the formation of an intermediate 8 with a Cu + and a series of intermediates such as CuL 2, Cu 2 L 2, Cu 3 L 2 (L stands for the ligand 4).
Figure S14. Emission intensity at 580 nm of 20 μm 4 in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution vs ph before and after addition of 100 μm Cu 2+ (λ ex = 510 nm). The ph of the solutions was adjusted by addition of 0.2 mol L -1 HCl (or 0.1 mol L 1 NaOH). Figure S15. Absorption spectra of 20 µm 4 upon the addition of 100µM CuCl 2, Cu(NO 3 ) 2, CuSO 4, respectively, suggesting that the anions have no influence on the response of 4 to Cu 2+.
Figure S16. Fluorescence spectra of 20 µm 4 upon addition of 0 20 µm Cu 2+ in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution (λ ex = 510 nm). Inset: Emission intensity at 580 nm as a function of Cu 2+ concentration.
Figure S17. Absorption spectra of 20 µm 4 in the presence of different metal ions (10 mm for Na +, K +, Mg 2+, Ca 2+, 25 μm for Fe 2+ and Cu +, and 100 µm for other metal ions) in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution. Figure S18. Fluorescence spectra of 20 µm 4 in the presence of different metal ions (10 mm for Na +, K +, Mg 2+, Ca 2+, 25 μm for Fe 2+ and Cu +, and 100 µm for other metal ions) in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution.
Figure S19. Confocal fluorescence, brightfield, and overlay images of MCF-7 cells. The cells were supplemented with 0 (a1-a3), 50 (b1-b3), 100 (c1-c3) or 200 (d1-d3) μm CuCl 2 in the growth media for 20 hours at 37 o C, and then were stained with 50 μm 4 for 30 min at 37 o C (λ ex = 543 nm). Figure S20. Cell viability values (%) estimated by trypan blue viability test. MCF-7 cells were cultured in the presence of 50 or 200 μm Cu 2+ for 20 h at 37 C and then incubated with 50 μm 4 for 30 min at 37 o C.
Part 2. Colour versions of Figures 2, 5, 7 and 8, Scheme 3, and Table 1 in Text. Figure 2. Distinct difference in Cu 2+ and Hg 2+ sensing behavior of 4 in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution. (a) Time course of 20 µm 4 upon gradual addition of Cu 2+ and Hg 2+. Absorbance at 555 nm (for Cu 2+ ) and 566 nm (for Hg 2+ ) was recorded respectively. Inset shows the fluorescent responses of 4 to 5 equiv. of Cu 2+ and Hg 2+ under 365 nm UV excitation. (b) Color changes of 20 µm 4 in the presence of different concentrations of Cu 2+ or Hg 2+. Figure 5. Time course of the response of 4 to different concentrations of Cu 2+. Absorbance at 555 nm was recorded in 20 μm 4 in CH 3 CN/HEPES (50 mm, ph 7.2, 3:7, v/v) solution.
Figure 7. Confocal fluorescence and brightfield images of HeLa cells. (a) Cells stained with 5 μm 4 for 10 min at 25 o C. (b) Cells supplemented with 50 μm CuCl 2 in the growth media for 20 hours at 37 o C and then incubated with 5 μm 4 for 10 min at 25 o C. (c) Brightfield image of cells shown in panel b. The overlay image of (b) and (c) is shown in (d) (λ ex = 543 nm). Figure 8. (a) Two-photon excited fluorescence image of Cu 2+ supplemented HeLa cells stained with 5 μm 4 for 10 min at 25 o C (λ ex = 880 nm). (b) Fluorescence intensity profile across a HeLa cell shown in (a).
Table 1. HOMO-1, HOMO, LUMO and LUMO+1 distributions of 1 and 4 calculated by DFT calculations.