Copyright WILEY-VCH Verlag GmbH & Co. KGaA, 69469 Weinheim ... · Figure S3. HRMS spectrum of...
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Copyright WILEY-VCH Verlag GmbH & Co. KGaA, 69469 Weinheim, Germany, 2016. Supporting Information Rational design of a water-soluble NIR AIEgen, and its applications for ultrafast wash-free cellular imaging and photodynamic cancer cell ablation Dong Wang, Huifang Su, Ryan T. K. Kwok, Xianglong Hu,* Hang Zou, Qianxin Luo, Michelle M. S. Lee, Wenhan Xu, Jacky W. Y. Lam, Ben Zhong Tang* Table of Contents Experimental procedures Table S1. Optical properties of AIEgens TVP and TTVP. Figure S1. 1 H NMR spectrum of TVP. Figure S2. 13 C NMR spectrum of TVP. Figure S3. HRMS spectrum of compound TVP. Figure S4. 1 H NMR spectrum of 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde. Figure S5. 1 H NMR spectrum of TTVP. Figure S6. 13 C NMR spectrum of TTVP. Figure S7. HRMS spectrum of compound TTVP. Figure S8. Particle size distributions of TVP aggregates. Figure S9. PL spectra of TTVP in solvents with different polarities. Figure S10. Confocal images of HeLa cells stained with TTVP for different time. Figure S11. Co-localization imaging of HeLa cells stained with TVP and DiO. Figure S12. Plasma membrane-imaging of other cells. Figure S13. Chemical trapping measurements of the 1 O2 quantum yield. Figure S14. Decomposition rates of ABDA with light irradiation under different conditions. Figure S15. Morphology change of plasma membrane upon light irradiation. Figure S16. Co-localization imaging of COS-7 cells stained with TTVP and DiO. Figure S17. PDT application of TTVP based on COS-7 cells. Electronic Supplementary Material (ESI) for Chemical Science. This journal is © The Royal Society of Chemistry 2018
Transcript of Copyright WILEY-VCH Verlag GmbH & Co. KGaA, 69469 Weinheim ... · Figure S3. HRMS spectrum of...
Copyright WILEY-VCH Verlag GmbH & Co. KGaA, 69469 Weinheim, Germany, 2016.
Supporting Information
Rational design of a water-soluble NIR AIEgen, and its applications for ultrafast
wash-free cellular imaging and photodynamic cancer cell ablation
Dong Wang, Huifang Su, Ryan T. K. Kwok, Xianglong Hu,* Hang Zou, Qianxin Luo,
Michelle M. S. Lee, Wenhan Xu, Jacky W. Y. Lam, Ben Zhong Tang*
Table of Contents
Experimental procedures
Table S1. Optical properties of AIEgens TVP and TTVP.
Figure S1. 1H NMR spectrum of TVP.
Figure S2. 13C NMR spectrum of TVP.
Figure S3. HRMS spectrum of compound TVP.
Figure S4. 1H NMR spectrum of 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde.
Figure S5. 1H NMR spectrum of TTVP.
Figure S6. 13C NMR spectrum of TTVP.
Figure S7. HRMS spectrum of compound TTVP.
Figure S8. Particle size distributions of TVP aggregates.
Figure S9. PL spectra of TTVP in solvents with different polarities.
Figure S10. Confocal images of HeLa cells stained with TTVP for different time.
Figure S11. Co-localization imaging of HeLa cells stained with TVP and DiO.
Figure S12. Plasma membrane-imaging of other cells.
Figure S13. Chemical trapping measurements of the 1O2 quantum yield.
Figure S14. Decomposition rates of ABDA with light irradiation under different conditions.
Figure S15. Morphology change of plasma membrane upon light irradiation.
Figure S16. Co-localization imaging of COS-7 cells stained with TTVP and DiO.
Figure S17. PDT application of TTVP based on COS-7 cells.
Electronic Supplementary Material (ESI) for Chemical Science.This journal is © The Royal Society of Chemistry 2018
Experimental procedures
Cell culture
HeLa cells were cultured in the MEM containing 10% FBS and antibiotics (100 units/mL penicillin
and 100 mg/mL streptomycin) in a 5% CO2 humidity incubator at 37 oC.
Confocal colocalization
After incubating HeLa cells with DiO at 37 oC for 10 min, TTVP was added into the culture, which
was then shaken for a few seconds at room temperature. The medium was then removed and the
cells were rinsed with PBS for three times and then imaged under confocal microscope. For TTVP,
the emission filter was 600-744 nm; for DiO, the emission filter was 490–600 nm.
Photostability
The TTVP-labelled HeLa cells were imaged by a confocal microscope (Zeiss laser scanning
confocal microscope LSM7 DUO) using ZEN 2009 software (Carl Zeiss). Conditions: for TTVP,
excitation wavelength: 488 nm; for DiO, excitation wavelength: 488nm (5% laser power). 600–740
nm; for BODIPY493/503 Green, the excitation was 488 nm and the emission filter was 510–553
nm.
Singlet-triplet energy gap calculations
The calculation of excited state was performed at TD-DFT CAM-B3LYP/6-31G(d) level. The
calculation of groud state and triplet state were performed at CAM-B3LYP/6-31G(d) level.
Table S1. Optical properties of AIEgens TVP and TTVP.
AIEgens λabs ε
(M-1 cm-1)
λem [nm] αAIE
(Iaggr, max/Isolu)
τ
[ns]f) [nm]a) Soln (ΦF)b,c) Aggr(ΦF)c,d) Solid (ΦF)c,e)
TVP 467 25070 619 (0.2%) 629 (4.4%) 657 (4.9%) 41.7 5.75
TTVP 480 33517 N/A 708 (1.7%) 705 (2.7%) 97.3 0.92
a)Absorption maximum in aqueous solutions; b)Emission maximum in aqueous
solutions (10 µM); c)Fluorescence quantum yield determined by a calibrated integrating
sphere; d)Emission maximum in aggregation state; e)Emission maximum in solid state; f)Fluorescence lifetime, measured under ambient conditions.
Figure S1. 1H NMR spectrum of TVP.
Figure S2. 13C NMR spectrum of TVP.
9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 ppm
2.4
38
2.4
57
2.4
75
3.1
15
3.3
96
3.4
50
3.4
71
3.4
91
4.5
90
4.6
08
4.6
26
6.9
29
6.9
50
7.1
04
7.1
23
7.1
43
7.1
62
7.1
80
7.3
38
7.3
60
7.3
79
7.3
99
7.6
33
7.6
55
8.0
09
8.0
50
8.2
22
8.2
39
9.0
33
9.0
49
1.8
3
9.1
7
2.3
9
1.9
1
2.3
2
6.1
6
5.3
3
1.9
7
0.9
9
1.9
1
1.8
9
170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm
24.1
84
38.8
74
39.0
82
39.2
91
39.5
00
39.7
08
39.9
17
40.1
26
52.4
20
56.3
23
61.7
67
120
.465
120
.636
123
.239
124
.554
125
.427
127
.971
129
.791
129
.870
141
.046
144
.222
146
.165
149
.542
153
.576
Figure S3. HRMS spectrum of compound TVP.
Figure S4. 1H NMR spectrum of 5-(4-(diphenylamino)phenyl)thiophene-2-
carbaldehyde.
10 9 8 7 6 5 4 3 2 1 0 ppm
7.0
50
7.0
67
7.0
72
7.0
89
7.1
07
7.1
25
7.1
27
7.1
46
7.1
49
7.2
76
7.2
94
7.2
97
7.3
03
7.3
16
7.5
07
7.5
29
7.7
03
7.7
13
9.8
53
7.8
8
5.0
3
2.0
0
1.0
0
0.9
2
Figure S5. 1H NMR spectrum of TTVP.
Figure S6. 13C NMR spectrum of TTVP.
12 11 10 9 8 7 6 5 4 3 2 1 0 ppm
2.5
14
2.5
34
2.5
44
2.5
54
2.5
65
2.5
74
2.5
94
3.2
03
3.3
02
3.3
06
3.3
10
3.3
14
3.3
18
3.5
18
3.5
30
3.5
40
3.5
49
3.5
61
4.5
80
4.5
99
4.6
19
4.8
84
7.0
18
7.0
23
7.0
35
7.0
40
7.0
81
7.0
92
7.0
95
7.0
99
7.1
09
7.1
15
7.2
98
7.3
06
7.3
18
7.3
37
7.3
92
7.4
02
7.4
77
7.4
87
7.5
82
7.5
86
7.5
98
7.6
03
8.1
25
8.1
31
8.1
43
8.1
71
8.7
71
8.7
88
2.0
8
9.0
0
2.0
3
2.1
6
9.2
4
4.0
4
1.0
6
0.9
8
1.9
3
2.8
4
1.8
9
190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 ppm
26.2
68
48.5
60
48.7
73
48.9
87
49.2
00
49.4
12
49.6
24
49.8
37
54.1
27
58.0
40
64.0
40
121
.712
123
.798
124
.902
125
.036
125
.156
126
.399
128
.117
128
.267
130
.812
135
.831
136
.717
140
.327
145
.258
148
.719
150
.186
150
.811
155
.986
Figure S7. HRMS spectrum of compound TTVP.
Figure S8. Particle size distributions of TVP aggregates in water/THF mixture with 90%
THF fraction.
Figure S9. a) PL spectra of TTVP in solvents with different polarities. Concentration:
0.5 μM; excitation wavelength: 480 nm. b) The data of emission maximum in (a).
Figure S10. Confocal images of HeLa cells stained with TTVP for different time.
Concentration: 500 nM. Scale bar = 20 µm. λex: 488 nm (1% laser power, 0.05 μW).
Figure S11. a) Confocal images and b) bright field of living HeLa cells after incubation
with TVP (500 nM) by the use of washing procedure after incubation for 3 seconds.
λex: 488 nm (1.2% laser power).
20 mm
a b
a b
Figure S12. Extension of the wash-free and plasma membrane-specific imaging
strategy to other cells. Confocal images of living a) 293T, b) HCC827, c) HCT116, and
d) MDCK2 cells after incubation with TTVP (500 nM) for an extremely short
incubation period (around 3 s). λex: 488 nm (1% laser power, 0.05 μW).
Figure S13. Chemical trapping measurements of the 1O2 quantum yield.
Photodegradation of ABDA with Rose Bengal (a), TTVP (d) and TVP (g). The
absorption peak area of Rose Bengal (b), TTVP (e) and TVP (h). The decomposition
rate constants of ABDA by Rose Bengal (c), TTVP (f) and TVP (I). To eliminate the
inner-filter effect, the absorption maxima were adjusted to ~0.2 OD. These
a b c
d e f
g h i
Rose Bengal
TTVP
TVP
0
6
min
0
6
min
0
6
min
Rose
Bengal
Rose
Bengal
TTVP TTVP
TVP TVP
Area = 9.875
Area = 25.911
Area = 19.712
Y = 0.00265 + 0.000977X
R2 = 0.99921
Y = 0.01908 + 0.00274X
R2 = 0.99609
Y = 0.0632 + 0.00128X
R2 = 0.98905
measurements were carried out under white light irradiation in DMSO/water (v:v) =
1/100. [ABDA] = 10 × [AIEgens or Rose Bengal], time interval for recording the UV-
vis spectra: 20 s.
Figure S14. Decomposition rates of ABDA with light irradiation under different
conditions, where A0 and A are the absorbance of ABDA at 378 nm. [TTVP] = 5×10-6
M, [ABDA] = 5×10-5 M, time interval for recording the UV-vis spectra: 60 s.
Figure S15. Bright-field of confocal images of HeLa cell stained with TTVP upon the
increase of irradiation time. λex: 488 nm (20% laser power, 0.925 μW).
Figure S16. Co-localization imaging of COS-7 cells stained with TTVP and DiO.
Confocal images of COS-7 cells stained with b) TTVP, c) CellMask Green, and d)
merged images of panels b) and c), and a) Bright-field. Concentrations: TTVP (0.5 μM).
The emission filter TTVP: 600-744 nm; the emission filter of CellMask Green: 490–
565 nm.
a b
e d c 20 mm
Figure S17. Cell viability of COS-7 cells stained with different concentrations of TTVP
in the absence or presence of white light irradiation for 10 min. Light power: 10
mW/cm2.
