Supporting Information Unimolecular Photoconversion of Multicolor Luminescence on Hierarchical Self-Assemblies Liangliang Zhu, Xin Li, Quan Zhang, Xing Ma, # Menghuan Li, # Huacheng Zhang, Zhong Luo, Hans Ågren, and Yanli Zhao *,,# Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University, Nanyang Link, Singapore 677 Department of Theoretical Chemistry and Biology, School of Biotechnology, KTH Royal Institute of Technology, SE-069 Stockholm, Sweden # School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore 69798 S
Table of Contents Conformational Geometries of Compound S Characterization Spectra of S5 Supplementary Optical Studies S7 TEM Images with a Relatively Large Scale S9 Supplementary Calculation Data for Molecular Dynamics Simulations S0 MTT Cytotoxic Study S Intracellular Micro-Spectroscopies Measured by Confocal Microscopes S Cell Imaging Observed from a Fluorescence Microscopy S8 Full Citation for Reference 6 S9 S
Conformational Geometries of Compound : Table S. Computational results for the transitions of compound in Z-form. Excited state Excitation energy MO composition Oscillator strength S. ev, 6 nm H-0 -> L+ ( 96%) f=.79 S.5 ev, 5 nm H- -> L+0 ( 96%) f=0.88 S.9 ev, 88 nm H-5 -> L+0 ( 6%) f=0.000 H- -> L+ ( 5%) S.5 ev, 7 nm H-0 -> L+0 ( 79%) f=0.0006 S 5.58 ev, 70 nm H- -> L+0 ( 77%) f=0.007 S 6.6 ev, 67 nm H-6 -> L+ ( %) f=0.00 H-0 -> L+5 ( 65%) S 7.76 ev, 60 nm H- -> L+ ( 8%) f=0.00 Table S. Computational results for the transitions of compound in E-form. Excited state Excitation energy MO composition Oscillator strength S.5 ev, 5 nm H- -> L+0 ( 96%) f=0.506 S.55 ev, 9 nm H-0 -> L+ ( 95%) f=0.90 S.9 ev, 88 nm H-6 -> L+0 ( 6%) f=0.00 H- -> L+ ( 5%) S.5 ev, 7 nm H-0 -> L+0 ( 79%) f=0.0007 S 5.58 ev, 70 nm H- -> L+0 ( 77%) f=0.00 S 6.6 ev, 67 nm H-5 -> L+ ( 5%) f=0.0599 H-0 -> L+ ( %) H-0 -> L+5 ( %) S 7.76 ev, 60 nm H- -> L+ ( 9%) f=0.7 S
Table S. Important frontier MOs of compound MO in Z-form in E-form HOMO- (H-) HOMO (H-0) LUMO (L+0) LUMO+ (L+) Figure S. Calculated excitation energy for the luminophores (-dimethylamino-cyanostilbene and -piperazinyl-,8-naphthalimide) of the Z-isomer and E-isomer in compounds. S
Characterization Spectra of : O HN N N O N NC R O () Br 0 O () N Br 0 N N O R N NC OH a b c N d e f g O NC () N N 0 h i j k O N O Figure S. Synthetic route for the preparation of the compound. The protons of the luminophors are defined alongside the structural formula. Figure S. H NMR spectra (00 MHz, DMSO-d 6, 98 K) of (a) at initial state, (b) after photoirradiation at 5 nm for 6 h, and (c) followed by heating over 00 ºC for 6 h. S5
Figure S. MS spectra (ESI) of (a) at initial state and (b) after sufficient photoirradiation. S6
Supplementary Optical Studies: 0% Intensity/a.u. 90% 0% ~ 90% 00 50 500 550 600 650 700 Figure S5. Emission spectra (λ ex = 65 nm) of R (black curves) in initial state and (blue curves) after sufficient photoirradiation in DMSO with different amount (0% ~ 90%) of water. Absorption Intensity/a.u. 5 6 7 8 50 00 50 500 550 Figure S6. The normalized absorption spectra of under different solvent conditions: in () n-hexane, () toluene, () THF, () ethylene glycol, (5) DMSO, (6) DMSO with 0% water, (7) DMSO with 90% water, and (8) DMSO with 90% water after sufficient photoirradiation. S7
Intensity/a.u. 6 R in n-hexane R in toluene R in THF R in ethylene glycol R in n-hexane R in toluene R in THF R in ethylene glycol 0 00 50 500 550 600 650 700 Figure S7. Emission spectra of R and R in solvents with different polarity. All the data were collected at the concentration of 0.0 mm at 98 K. 0 (a) Intensity/a.u. 5 0 5 0 5 0 5 5 6 7 8 9 0 0-5 00 50 500 550 600 650 700 Figure S8. (a) Emission spectra (λ ex = 65 nm) of in DMSO with ) 0%, ) 0%, ) 0%, ) 0%, 5) 50%, 6) 60%, 7) 70%, 8) 80%, 9) 90% and 0) 99% of water; (b) Photo images of the corresponding states in (a) under a UV light ( λ = 65 nm). S8
(a) Intensity/a.u. 0 0 00 80 60 0 0 5 6 7 8 9 (b) 0 Intensity/a.u. 00 80 60 0 0 5 6 7 8 9 0 0 00 50 500 550 600 650 700 00 50 500 550 600 650 700 Figure S9. Emission spectra (λ ex = 65 nm) of (a) R and (b) a mixture of R/R (: molar ratio) in DMSO with ) 0%, ) 0%, ) 0%, ) 0%, 5) 50%, 6) 60%, 7) 70%, 8) 80%, 9) 90% and 0) 99% of water. The emissions of R in the same condition are still quenched relative to R. TEM Images with a Relatively Large Scale: Figure S0. TEM images of prepared from (a) DMSO with 90% water, (b) DMSO with 0% water and (c) DMSO with 90% water after sufficient photoirradiation with a relatively large scale. The TEM copper-grids were subsequently stained with a % aqueous sodium phosphotungstate solution and allowed to air-dry. Some helical twists were highlighted by arrows. S9
Supplementary Calculation Data for Molecular Dynamics Simulations: Figure S. Calculated averaged electronic circular dichroism spectra of in Z- and E-forms in DMSO with 90% water. Figure S. Snapshot of transient self-assembled conformations of (a) R and (b) R under the environment of DMSO with 90% water. S0
Figure S. Solvent accessible surface (SAS) area of the self-assemblies of in () Z-form and () E-form in DMSO/H O (9:) solution. Open circles represent SAS areas computed from snapshots, while full lines show cubic spline smoothing. Figure S. Order parameter of cyanostilbene and naphthalimide subunits in the self-assemblies of in () Z-form and () E-form in DMSO/H O (9:) solution. Open circles represent order parameters computed from snapshots, while full lines show cubic spline smoothing. S
Intensity/a.u. 00 50 500 550 600 650 700 Figure S5. Emission spectra (λ ex = 65 nm) of in DMSO with 90% water at () initial state and () after sufficient photoirradiation at 5 nm and () after heating over 00 ºC overnight and then cooling to room temperature. MTT Cytotoxic Study: 00 Viability/% 80 60 0 0 0 6 5 50 500 000 Concentration of the dyad/nm Figure S6. Cell viability (%) estimated by a MTT proliferation test versus incubation concentrations of () and () after sufficient photoirradiation. The cells were cultured in the presence of different concentrations of the dyad at 7 ºC for h. S
Intracellular Micro-Spectroscopies Measured by Confocal Microscopes: Figure S7. Confocal fluorescence images of HeLa cells incubated with compound in initial state in DMEM medium containing 5% DMSO/PBS (ph 7., :9, v/v) at 7 C for h. The images were collected under different channels (readout wavelengths). The intensity values along the line highlighted were collected and processed (see Figure S8). (a) (b) Intensity/a.u 5 6 7 0.00000 0.00005 0.00000 0.00005 0.00000 0.00005 Position Average Intensity/a.u. 50 500 550 600 650 Figure S8. (a) The intensity values of different channels (corresponding to the images in Figure S7) along the line highlighted in Figure S7. (b) Plot of the average intensities of different channels in (a) against the corresponding central readout wavelengths. S
Figure S9. Confocal fluorescence images of HeLa cells incubated with compound in the state after photoirradiation for h in DMEM medium containing 5% DMSO/PBS (ph 7., :9, v/v) at 7 C for h. The images were collected under different channels (readout wavelengths). The intensity values along the line highlighted were collected and processed (see Figure S0). (a) Intensity/a.u 5 6 7 0.00000 0.00008 0.000056 0.00006 0.00007 Position (b) Average Intensity/a.u. 50 500 550 600 650 Figure S0. (a) The intensity values of different channels (corresponding to the images in Figure S9) along the line highlighted in Figure S9. (b) Plot of the average intensities of different channels in (a) against the corresponding central readout wavelengths. S
Figure S. Confocal fluorescence images of HeLa cells incubated with compound in the state after photoirradiation for 8h in DMEM medium containing 5% DMSO/PBS (ph 7., :9, v/v) at 7 C for h. The images were collected under different channels (readout wavelengths). The intensity values along the line highlighted were collected and processed (see Figure S). (a) (b) Intensity/a.u 5 6 7 0.0000 0.00000 0.00008 0.000056 0.00006 Position Average Intensity/a.u. 50 500 550 600 650 Figure S. (a) The intensity values of different channels (corresponding to the images in Figure S) along the line highlighted in Figure S. (b) Plot of the average intensities of different channels in (a) against the corresponding central readout wavelengths. S5
Figure S. Confocal fluorescence images of HeLa cells incubated with compound R in DMEM medium containing 5% DMSO/PBS (ph 7., :9, v/v) at 7 C for h. The images were collected under different channels (readout wavelengths). The intensity values along the line highlighted were collected and processed (see Figure S). (a) Intensity/a.u 5 6 7 0.00000 0.00005 0.00000 0.00005 0.000050 Position (b) Average Intensity/a.u. 50 500 550 600 650 Figure S. (a) The intensity values of different channels (corresponding to the images in Figure S) along the line highlighted in Figure S. (b) Plot of the average intensities of different channels in (a) against the corresponding central readout wavelengths. S6
90 80 Intensity/a.u 70 60 50 0 0 0 0 0 50 500 550 600 650 700 Figure S5. Emission spectra (λ ex = 05 nm) of at 7 ºC in DMSO with 99% PBS solution at () initial state and after photoirradiation at 5 nm for () h and () 8 h. 0 6 Intensity/a.u. 8 0 00 50 500 550 600 650 Figure S6. Emission spectra (λ ex = 65 nm) of in DMSO with 99% PBS solution at initial state () before and () after exposing at 7 ºC for h; Emission spectra (λ ex = 65 nm) of in DMSO with 99% PBS solution at photostationary state () before and () after exposing at 7 ºC for h. The comparison of these curves shows that effective isomerization of this compound would not take place under this condition. S7
Cell Imaging Observed from a Fluorescence Microscopy: Figure S7. Fluorescent images of HeLa cells incubated with (a) compound and (b) compound after sufficient photoirradiation in DMEM medium containing 0% DMSO/PBS (ph 7., :9, v/v) at 7 C for h. For each panel, the images from left to right were collected under bright field, FITC channel (~50 nm), and DAPI channel (~50 nm). Figure S8. Comparison between bond lengths, angles, and cosine of dihedral angles of from quantum mechanical (QM) calculations and from molecular mechanical (MM) force field calculations. S8
Full Citation for Reference 6: (6) Frisch, M. J.; Trucks, G. W.; Schlegel, H. B.; Scuseria, G. E.; Robb, M. A.; Cheeseman, J. R.; Scalmani, G.; Barone, V.; Mennucci, B.; Petersson, G. A.; Nakatsuji, H.; Caricato, M.; Li, X.; Hratchian, H. P.; Izmaylov, A. F.; Bloino, J.; Zheng, G.; Sonnenberg, J. L.; Hada, M.; Ehara, M.; Toyota, K.; Fukuda, R.; Hasegawa, J.; Ishida, M.; Nakajima, T.; Honda, Y.; Kitao, O.; Nakai, H.; Vreven, T.; Montgomery Jr., J. A.; Peralta, J. E.; Ogliaro, F.; Bearpark, M.; Heyd, J. J.; Brothers, E.; Kudin, K. N.; Staroverov, V. N.; Kobayashi, R.; Normand, J.; Raghavachari, K.; Rendell, A.; Burant, J. C.; Iyengar, S. S.; Tomasi, J.; Cossi, M.; Rega, N.; Millam, J. M.; Klene, M.; Knox, J. E.; Cross, J. B.; Bakken, V.; Adamo, C.; Jaramillo, J.; Gomperts, R.; Stratmann, R. E.; Yazyev, O.; Austin, A. J.; Cammi, R.; Pomelli, C.; Ochterski, J. W.; Martin, R. L.; Morokuma, K.; Zakrzewski, V. G.; Voth, G. A.; Salvador, P.; Dannenberg, J. J.; Dapprich, S.; Daniels, A. D.; Farkas, O.; Foresman, J. B.; Ortiz, J. V.; Cioslowski, J.; Fox, D. J. Gaussian 09, Revision A., Gaussian, Inc.: Wallingford CT, 009. S9