Supporting Information Aggregation-Induced-Emission Materials with Different Electric Charges as an Artificial Tongue: Design, Construction, and Assembly with Various Pathogenic Bacteria for Effective Bacterial Imaging and Discrimination Guang-jian Liu, a Sheng-nan Tian, b,c Cui-yun Li, a Guo-wen Xing* a and Lei Zhou* b,c a. College of Chemistry, Beijing Normal University, Beijing 100875, China. b. National Key Laboratory of Biochemical Engineering, PLA Key Laboratory of Biopharmaceutical Production & Formulation Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China. c. State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China. *E-mail: gwxing@bnu.edu.cn, ammszhoulei@aliyun.com S-1
Scheme S1. Synthetic route to compounds 1 and 2. Synthesis of 1-[(4-bromomethyl)phenyl]-1,2,2-triphenylethene (1) and 1,2-bis[4- (bromomethyl)phenyl]-1,2-diphenylethene (2). The synthesis of 1 and 2 was according to the procedures of previous literature. Zinc dust (1.7344 g, 26. 6mmol), benzophenone (431.4 mg, 2.367 mmol)and 4-methylbenzophenone (981.2 mg, 5 mmol )were added into a 250-mL two-necked roundbottom flask with a magnetic stirrer and a water condenser. The flask was degassed and flushed with argon three times, after which 30 ml THF was injected. The mixture was stirred at -50 C for 10 min, then titanium tetrachloride (1.13mL, 10.31mmol) was added slowly. After stirred at room temperature for 30 min, the mixture was refluxed at 70 C overnight. Then the reaction was quenched with 10% potassium carbonate (K2CO3) aqueous solution and filtrated. The filtrate was extracted with CH2Cl2 twice and the combined organic fractions were dried with anhydrous magnesium sulfate (MgSO4), filtrated and concentrated under reduce pressure. The residue was purified by column chromatography to give a mixture of 1,2-diphenyl-1,2-di-p-tolylethene and 1,2,2-triphenyl-1-p-tolylethene which could not be easily separated due to the similar Rf. Afterwards, N-bromosuccinimide (NBS, 900mg, 5mmol), benzoyl peroxide (BPO, 20mg) and the product mixture (1 g) of 1,2-diphenyl-1,2-di-p-tolylethene and 1,2,2-triphenyl-1-p-tolylethene were added to a 100-mL round-bottom flask fitted with a condenser. Then CCl4 (30mL) was added under argon atmosphere and the reaction was heated to 70 C on an oil bath for 12 h. After the mixture was cooled to room temperature and filtered to remove the produced succinimide, the filtrate was evaporated under vacuum. The crude product was purified by column chromatography affording pure product 1 and 2. Compound 1. 1 H NMR (400 MHz, CDCl3) δ 7.15 7.07 (m, 11H), 7.05 6.96 (m, 8H), 4.42 (s, 2H). Compound 2. 1 H NMR (400 MHz) δ 7.15 7.08 (m, 10H), 7.04 6.94 (m, 8H), 4.42 (d, J = 6.4 Hz, 4H). The spectroscopic data coincide with the previous report. [ACS Appl. Mater. Interfaces, 2013, 5, 4613 4616; J. Am. Chem. Soc., 2013, 135, 62 65] S-2
Figure S1. Fluorescence spectra of (a) TPEMN (b) TPEDN at different concentrations in physiological saline. TPEMN: λex = 340 nm; TPEDN: λex = 337 nm. Figure S2. Fluorescence spectra of aqueous suspensions of (a) TPEMC (b) TPEDC (c) TPEMA (d) TPEDA at different concentrations. TPEMC: λex = 367 nm; TPEDC: λex = 350 nm; TPEMA: λex = 345 nm; TPEDA: λex = 364 nm S-3
Figure S3. Hydrodynamic diameters and distribution of TPEMN, TPEMA, TPEMC as determined by dynamic light scattering (DLS) at the concentration of 24 μg/ml. Figure S4. LSCM images of Gram-negative E.coli after incubation with different concentrations of TPEMN for 1 min, 3 min, 5 min, 8 min and 10 min following described procedures of LIM and LIM- LSCM observation. Scale bar: 10 μm. Figure S5. Fluorescence spectra of Gram-negative E.coli suspensions with a concentration of 10 8 CFU/mL after incubation with different concentrations of TPEMN for (a) 1 min, (b) 3 min, (c) 5 min,(d) 8 min and (e) 10 min following described procedures of LIM and LIM-fluorescent spectrum measurement. λex = 332 nm. S-4
Figure S6. LSCM images of Gram-positive S.aureus after incubation with different concentrations of TPEMN for 1 min, 3 min, 5 min, 8 min and 10 min following described procedures of LIM and LIM- LSCM observation. Scale bar: 10 μm. Figure S7. Fluorescence spectra of Gram-positive S.aureus suspensions with a concentration of 10 8 CFU/mL after incubation with different concentrations of TPEMN for (a) 1 min, (b) 3 min, (c) 5 min, (d) 8 min and (e) 10 min following described procedures of LIM and LIM-fluorescent spectrum measurement. λex = 332 nm, slitex = slitem = 5 nm. S-5
Figure S8. LSCM images of Gram-negative E.coli after incubation with 24 μg/ml of TPEMC, TPEDC, TPEMA, TPEDA, TPEDN for 1 min, 3 min, 5 min, 8 min and 10 min following described procedures of LIM and LIM-LSCM observation. Scale bar: 10 μm. Figure S9. LSCM images of Gram-positive S.aureus after incubation with 24 μg/ml of TPEMC, TPEDC, TPEMA, TPEDA, TPEDN for 1 min, 3 min, 5 min, 8 min and 10 min following described procedures of LIM and LIM-LSCM observation. Scale bar: 10 μm. S-6
Figure S10. Fluorescence images and bright field of Gram-positive bacteria (L. monocytogenes, S. aureus, B. subtilis) after incubation with 24 μg/ml of probes (TPEMC, TPEDC, TPEMA, TPEDA, TPEMN and TPEDN) for 5 min following described procedures of LIM and LIM-LSCM observation. S-7
Figure S11. Fluorescence response patterns ((I I0)/I0) obtained by six probes (24 μg/ml) treated with eight kinds of bacteria (10 8 cfu/ml) according to the described procedures. Each value is the average of three independent measurements; each error bar shows the standard deviation of these measurements. The excitation wavelengths are 337 nm, 337 nm, 362 nm, 369 nm, 332 nm and 334 nm for each TPE probe, respectively; the emission wavelengths are listed in the figure. I0 is the corresponding fluorescence intensity of each TPE material in physiological saline. Figure S12. (a) Fluorescence spectrum of TPEMN (24 μg/ml) in physiological saline and fluorescence spectra of bacteria suspensions with a concentration of 10 8 cfu/ml incubated with 24 μg/ml TPEMN for 5 min followed by centrifugation (7000 rpm, 5 min) and resuspension. (b) Fluorescence spectrum of TPEDN (24 μg/ml) in physiological saline and fluorescence spectra of bacteria suspensions with a concentration of 10 8 CFU/mL incubated with 24 μg/ml TPEDN for 5 min followed by centrifugation (7000 rpm, 5 min) and resuspension. Slitex = slitem = 5 nm. S-8
Figure S13. (a) 3D canonical score plot and (b) 2D canonical score plot for eight kinds of bacteria at 10 8 cfu/ml analyzed by LDA without TPEMC and TPEDC (three replicates for each kind of bacteria). Cross-validation showed 100% classification accuracy. S-9
NMR spectra Figure S14. 1 H NMR of compound 1. Figure S15. 1 H NMR of compound 2. S-10
Figure S16. 1 H NMR of compound TPEMA. Figure S17. 1 H NMR of compound TPEDA. S-11
Figure S18. 1 H NMR of compound TPEMC. Figure S19. 1 H NMR of compound TPEDC. S-12
Figure S20. 1 H NMR of compound TPEMN. Figure S21. 1 H NMR of compound TPEDN. S-13
Figure S22. 13 C NMR of compound TPEDN. Mass spectra Figure S23. HRMS of compound TPEMA. S-14
Figure S24. HRMS of compound TPEDA. Figure S25. HRMS of compound TPEMC. S-15
Figure S26. HRMS of compound TPEDC. Figure S27. HRMS of compound TPEMN. S-16
Figure S28. HRMS of compound TPEDN. S-17