本研究以氯化鐵氧化聚合方法成功合成出以疏水性聚噻吩為主體（作為發光骨架）及親水性的咪唑基團組成的水溶性共軛高分子。聚噻吩具有良好的光、電特性，可於巨觀與微觀條件下觀察螢光表現，而具有親水性的咪唑側鏈作為氣體響應性的官能基團，可以在水溶液中自組裝形成奈米微胞，並且經過二氧化碳與氮氣交替鼓泡入水溶液中，可以展現出具有回復性質的顆粒粒徑、表面電位與螢光特性，並且此一回復性質可以持續多次仍維持高穩定性。在體外細胞實驗中，pH 7.4、37°C生理條件下，在正常細胞與癌細胞中均展現出低細胞毒性；在細胞的螢光影像與流式細胞儀的研究表明，在二氧化碳鼓泡後，能展現出更高的細胞螢光強度與細胞攝取率，證實本實驗的實驗材料在未來有極佳潛力應用於生醫螢光探針等領域。

In this study, a water-soluble conjugated polymer composed of a hydrophobic polythiophene as the main body (as a light-emitting skeleton) and a hydrophilic imidazole group was successfully synthesized by the oxidative polymerization method of ferric chloride. Polythiophene has good optical and electrical properties, and its fluorescence performance can be observed under macro and micro conditions. The hydrophilic imidazole side chain, as a gas-responsive functional group, can self-assemble in aqueous solution to form nano-microstructures. After carbon dioxide and nitrogen are alternately bubbled into the aqueous solution, it can exhibit particle size, zeta potential and fluorescence characteristics with recovery properties, and this recovery property can continue for many times and still maintain high stability.
In the in vitro cell experiment, under physiological conditions of pH 7.4 and 37°C, it showed low cytotoxicity in normal cells and cancer cells. Fluorescence images and flow cytometry showed that after carbon dioxide bubbling can show higher cell fluorescence intensity and cell uptake rate, it confirmed that the experimental materials in this research have great potential to be used in the fields of biomedical fluorescent probes in the future.

1. Wang, H., Vagin, S. I., Rieger, B., & Meldrum, A. (2020). An ultrasensitive fluorescent paper-based CO2 sensor. ACS applied materials & interfaces, 12(18), 20507-20513.
2. Yuan, H., Fan, Y., Xing, C., Niu, R., Chai, R., Zhan, Y., ... & Xu, J. (2016). Conjugated polymer-based hybrid materials for turn-on detection of CO2 in plant photosynthesis. Analytical chemistry, 88(12), 6593-6597.
3. Wang, K., Amin, K., An, Z., Cai, Z., Chen, H., Chen, H., ... & Tang, B. Z. (2020). Advanced functional polymer materials. Materials Chemistry Frontiers, 4(7), 1803-1915.
4. Xu, X., Liu, R., & Li, L. (2015). Nanoparticles made of π-conjugated compounds targeted for chemical and biological applications. Chemical Communications, 51(94), 16733-16749.
5. Tuncel, D. (2019). π-Conjugated nanostructured materials: preparation, properties and photonic applications. Nanoscale Advances, 1(1), 19-33.
6. Wang, Y., Pei, Z., Feng, W., & Pei, Y. (2019). Stimuli-responsive supramolecular nano-systems based on pillar [n] arenes and their related applications. Journal of Materials Chemistry B, 7(48), 7656-7675.
7. Zhang, W., & Gao, C. (2017). Morphology transformation of self-assembled organic nanomaterials in aqueous solution induced by stimuli-triggered chemical structure changes. Journal of Materials Chemistry A, 5(31), 16059-16104.
8. Wang, H., Zhou, L., Zhou, C., Zhao, W., Wang, J., Liu, L., ... & Wang, Y. (2017). Preparation of gemini surfactant/conjugated polymer aggregates for enhanced fluorescence and bioimaging application. ACS applied materials & interfaces, 9(28), 23544-23554.
9. Yan, Q., & Zhao, Y. (2014). Block copolymer self-assembly controlled by the “green” gas stimulus of carbon dioxide. Chemical communications, 50(79), 11631-11641.
10. Varnava, C. K., & Patrickios, C. S. (2020). Polymer networks one hundred years after the macromolecular hypothesis: A tutorial review. Polymer, 123322.
11. Verpaalen, R. C., Engels, T., Schenning, A. P., & Debije, M. G. (2020). Stimuli-Responsive Shape Changing Commodity Polymer Composites and Bilayers. ACS Applied Materials & Interfaces, 12(35), 38829-38844.
12. Criado-Gonzalez, M., Dominguez-Alfaro, A., Lopez-Larrea, N., Alegret, N., & Mecerreyes, D. (2021). Additive Manufacturing of Conducting Polymers: Recent Advances, Challenges, and Opportunities. ACS Applied Polymer Materials.
13. Rother, M., Nussbaumer, M. G., Renggli, K., & Bruns, N. (2016). Protein cages and synthetic polymers: a fruitful symbiosis for drug delivery applications, bionanotechnology and materials science. Chemical Society Reviews, 45(22), 6213-6249.
14. Baek, P., Voorhaar, L., Barker, D., & Travas-Sejdic, J. (2018). Molecular approach to conjugated polymers with biomimetic properties. Accounts of chemical research, 51(7), 1581-1589.
15. Zhan, H., Chen, J., Zhang, C., Zhang, J., & Fan, L. J. (2021). Design, Synthesis, and Adhesion of Fluorescent Conjugated Polymers with Pendant Catechol Groups. ACS Applied Polymer Materials.
16. McQuade, D. T., Pullen, A. E., & Swager, T. M. (2000). Conjugated polymer-based chemical sensors. Chemical Reviews, 100(7), 2537-2574.
17. Feng, L., Zhu, C., Yuan, H., Liu, L., Lv, F., & Wang, S. (2013). Conjugated polymer nanoparticles: preparation, properties, functionalization and biological applications. Chemical Society Reviews, 42(16), 6620-6633.
18. Zhu, C., Liu, L., Yang, Q., Lv, F., & Wang, S. (2012). Water-soluble conjugated polymers for imaging, diagnosis, and therapy. Chemical reviews, 112(8), 4687-4735.
19. Zhou, L., Lv, F., Liu, L., & Wang, S. (2019). Water-soluble conjugated organic molecules as optical and electrochemical materials for interdisciplinary biological applications. Accounts of chemical research, 52(11), 3211-3222.
20. Yin, C., Song, W., Jiang, R., Lu, X., Hu, W., Shen, Q., ... & Huang, W. (2014). Oligo (p-phenyleneethynylene) embedded amphiphiles: synthesis, photophysical properties and self-assembled nanoparticles with high structural stability and photostability for cell imaging. Polymer Chemistry, 5(19), 5598-5608.
21. Arumugam, S., Vutukuri, D. R., Thayumanavan, S., & Ramamurthy, V. (2005). Amphiphilic homopolymer as a reaction medium in water: product selectivity within polymeric nanopockets. Journal of the American Chemical Society, 127(38), 13200-13206.
22. Liu, L., Yu, M., Duan, X., & Wang, S. (2010). Conjugated polymers as multifunctional biomedical platforms: Anticancer activity and apoptosis imaging. Journal of Materials Chemistry, 20(33), 6942-6947.
23. Lan, M., Liu, W., Ge, J., Wu, J., Wang, H., Zhang, W., ... & Wang, P. (2012). A polythiophene-derived ratiometric fluorescent sensor for highly sensitive determination of carbenicillin in aqueous solution. Chemical Communications, 48(54), 6818-6820.
24. Niemi, V. M., Knuuttila, P., Österholm, J. E., & Korvola, J. (1992). Polymerization of 3-alkylthiophenes with FeCl3. Polymer, 33(7), 1559-1562.
25. Liu, B., Wang, S., Bazan, G. C., & Mikhailovsky, A. (2003). Shape-adaptable water-soluble conjugated polymers. Journal of the American Chemical Society, 125(44), 13306-13307.
26. Feng, X., Yang, G., Liu, L., Lv, F., Yang, Q., Wang, S., & Zhu, D. (2012). A convenient preparation of multi‐spectral microparticles by bacteria‐mediated assemblies of conjugated polymer nanoparticles for cell imaging and barcoding. Advanced Materials, 24(5), 637-641.
27. Moon, J. H., McDaniel, W., MacLean, P., & Hancock, L. F. (2007). Live‐cell‐permeable poly (p‐phenylene ethynylene). Angewandte Chemie, 119(43), 8371-8373.
28. Wang, B., Yuan, H., Zhu, C., Yang, Q., Lv, F., Liu, L., & Wang, S. (2012). Polymer-drug conjugates for intracellar molecule-targeted photoinduced inactivation of protein and growth inhibition of cancer cells. Scientific reports, 2(1), 1-8.
29. Tu, Y., Zhou, Y., Zhang, D., Yang, J., Li, X., Ji, K., ... & Zhang, Q. (2021). Light-Induced Reactive Oxygen Species (ROS) Generator for Tumor Therapy through an ROS Burst in Mitochondria and AKT-Inactivation-Induced Apoptosis. ACS Applied Bio Materials.
30. Wu, C., Jin, Y., Schneider, T., Burnham, D. R., Smith, P. B., & Chiu, D. T. (2010). Ultrabright and bioorthogonal labeling of cellular targets using semiconducting polymer dots and click chemistry. Angewandte Chemie International Edition, 49(49), 9436-9440.
31. Wu, C., Hansen, S. J., Hou, Q., Yu, J., Zeigler, M., Jin, Y., ... & Chiu, D. T. (2011). Design of highly emissive polymer dot bioconjugates for in vivo tumor targeting. Angewandte Chemie, 123(15), 3492-3496.
32. Howes, P., Green, M., Levitt, J., Suhling, K., & Hughes, M. (2010). Phospholipid encapsulated semiconducting polymer nanoparticles: their use in cell imaging and protein attachment. Journal of the American Chemical Society, 132(11), 3989-3996.
33. Xing, C., Xu, Q., Tang, H., Liu, L., & Wang, S. (2009). Conjugated polymer/porphyrin complexes for efficient energy transfer and improving light-activated antibacterial activity. Journal of the American Chemical Society, 131(36), 13117-13124.
34. Yang, G., Liu, L., Yang, Q., Lv, F., & Wang, S. (2012). A multifunctional cationic pentathiophene: Synthesis, organelle‐selective imaging, and anticancer activity. Advanced Functional Materials, 22(4), 736-743.
35. Hu, L., Wan, Y., Zhang, Q., & Serpe, M. J. (2020). Harnessing the power of stimuli‐responsive polymers for actuation. Advanced Functional Materials, 30(2), 1903471.
36. Hu, L., Zhang, Q., Li, X., & Serpe, M. J. (2019). Stimuli-responsive polymers for sensing and actuation. Materials Horizons, 6(9), 1774-1793.
37. Bauri, K., Nandi, M., & De, P. (2018). Amino acid-derived stimuli-responsive polymers and their applications. Polymer Chemistry, 9(11), 1257-1287.
38. Lugger, S. J., Houben, S. J., Foelen, Y., Debije, M. G., Schenning, A. P., & Mulder, D. J. (2021). Hydrogen-Bonded Supramolecular Liquid Crystal Polymers: Smart Materials with Stimuli-Responsive, Self-Healing, and Recyclable Properties. Chemical Reviews.
39. Li, C., Wang, K., Li, J., & Zhang, Q. (2020). Recent progress in stimulus-responsive two-dimensional metal–organic frameworks. ACS Materials Letters, 2(7), 779-797.
40. Lin, X., Wang, X., Zeng, L., Wu, Z. L., Guo, H., & Hourdet, D. (2021). Stimuli-Responsive Toughening of Hydrogels. Chemistry of Materials.
41. Korde, J. M., & Kandasubramanian, B. (2019). Fundamentals and effects of biomimicking stimuli-responsive polymers for engineering functions. Industrial & Engineering Chemistry Research, 58(23), 9709-9757.
42. Yan, Y., Sun, N., Li, F., Jia, X., Wang, C., & Chao, D. (2017). Multiple stimuli-responsive fluorescence behavior of novel polyamic acid bearing oligoaniline, triphenylamine, and fluorene groups. ACS applied materials & interfaces, 9(7), 6497-6503.
43. Schmidt, M., Franken, A., Wilms, D., Fehm, T., Neubauer, H. J., & Schmidt, S. (2021). Selective Adhesion and Switchable Release of Breast Cancer Cells via Hyaluronic Acid Functionalized Dual Stimuli-Responsive Microgel Films. ACS Applied Bio Materials, 4(8), 6371-6380.
44. Huang, Y., Xue, Z., & Zeng, S. (2020). Hollow mesoporous Bi@ PEG-FA nanoshell as a novel dual-stimuli-responsive nanocarrier for synergistic chemo-photothermal cancer therapy. ACS Applied Materials & Interfaces, 12(28), 31172-31181.
45. Sikder, A., Chaudhuri, A., Mondal, S., & Singh, N. P. (2021). Recent Advances on Stimuli-Responsive Combination Therapy against Multidrug-Resistant Bacteria and Biofilm. ACS Applied Bio Materials.
46. Ganta, S., & Chand, D. K. (2017). Multi-Stimuli-Responsive Metallogel Molded from a Pd2 L 4-Type Coordination Cage: Selective Removal of Anionic Dyes. Inorganic chemistry, 57(7), 3634-3645.
47. Lin, P., Chen, H., Li, A., Zhuang, H., Chen, Z., Xie, Y., ... & Cheng, Z. (2020). Bioinspired Multiple Stimuli-Responsive Optical Microcapsules Enabled by Microfluidics. ACS Applied Materials & Interfaces, 12(41), 46788-46796.
48. Li, L., Yang, Z., & Chen, X. (2020). Recent Advances in Stimuli-Responsive Platforms for Cancer Immunotherapy. Accounts of Chemical Research, 53(10), 2044-2054.
49. Hu, X., Zhang, Y., Xie, Z., Jing, X., Bellotti, A., & Gu, Z. (2017). Stimuli-responsive polymersomes for biomedical applications. Biomacromolecules, 18(3), 649-673.
50. Chen, L. J., & Yang, H. B. (2018). Construction of stimuli-responsive functional materials via hierarchical self-assembly involving coordination interactions. Accounts of chemical research, 51(11), 2699-2710.
51. Mai, Z., Li, H., Gao, Y., Niu, Y., Li, Y., de Rooij, N. F., ... & Zhou, G. (2020). Synergy of CO 2-response and aggregation induced emission in a small molecule: renewable liquid and solid CO 2 chemosensors with high sensitivity and visibility. Analyst, 145(10), 3528-3534.
52. Yang, G., Hu, R., Ding, H. M., Kochovski, Z., Mei, S., Lu, Y., ... & Jiang, M. (2018). CO 2-switchable response of protein microtubules: behaviour and mechanism. Materials Chemistry Frontiers, 2(9), 1642-1646.
53. Yan, Q., & Zhao, Y. (2014). Block copolymer self-assembly controlled by the “green” gas stimulus of carbon dioxide. Chemical communications, 50(79), 11631-11641.
54. Jiang, H., Wang, E., & Wang, J. (2015). Gas-breathing polymer film for constructing switchable ionic diodes. RSC Advances, 5(45), 35622-35630.
55. Mabire, A. B., Brouard, Q., Pitto-Barry, A., Williams, R. J., Willcock, H., Kirby, N., ... & O'Reilly, R. K. (2016). CO 2/pH-responsive particles with built-in fluorescence read-out. Polymer Chemistry, 7(38), 5943-5948.
56. Terai, T., & Nagano, T. (2008). Fluorescent probes for bioimaging applications. Current opinion in chemical biology, 12(5), 515-521.
57. Goto, Y., Matsuno, R., Konno, T., Takai, M., & Ishihara, K. (2008). Artificial cell membrane-covered nanoparticles embedding quantum dots as stable and highly sensitive fluorescence bioimaging probes. Biomacromolecules, 9(11), 3252-3257.
58. Maestro, L. M., Rodríguez, E. M., Rodríguez, F. S., la Cruz, M. I. D., Juarranz, A., Naccache, R., ... & Solé, J. G. (2010). CdSe quantum dots for two-photon fluorescence thermal imaging. Nano letters, 10(12), 5109-5115.
59. Cai, L., Li, H., Yu, X., Wu, L., Wei, X., James, T. D., & Huang, C. (2021). Green Fluorescent Protein GFP-Chromophore-Based Probe for the Detection of Mitochondrial Viscosity in Living Cells. ACS Applied Bio Materials, 4(3), 2128-2134.
60. Zhang, J., Fu, Y., & Mahdavi, F. (2012). Bimetallic nanoshells for metal-enhanced fluorescence with broad band fluorophores. The Journal of Physical Chemistry C, 116(45), 24224-24232.
61. Cheng, C. C., Lai, Y. C., Shieh, Y. T., Chang, Y. H., Lee, A. W., Chen, J. K., ... & Lai, J. Y. (2020). CO2-Responsive Water-Soluble Conjugated Polymers for In Vitro and In Vivo Biological Imaging. Biomacromolecules, 21(12), 5282-5291.
62. Liu, Y., Liu, J., Chen, D., Wang, X., Liu, Z., Liu, H., ... & Zou, Y. (2019). Quinoxaline-based semiconducting polymer dots for in vivo NIR-II fluorescence imaging. Macromolecules, 52(15), 5735-5740.
63. Lv, Y., Liu, M., Zhang, Y., Wang, X., Zhang, F., Li, F., ... & Tian, Z. (2018). Cancer cell membrane-biomimetic nanoprobes with two-photon excitation and near-infrared emission for intravital tumor fluorescence imaging. ACS nano, 12(2), 1350-1358.
64. Matsuzaki, H., Kamiya, M., Iwatate, R. J., Asanuma, D., Watanabe, T., & Urano, Y. (2016). Novel hexosaminidase-targeting fluorescence probe for visualizing human colorectal cancer. Bioconjugate chemistry, 27(4), 973-981.
65. Grynkiewicz, G., Poenie, M., & Tsien, R. Y. (1985). A new generation of Ca2+ indicators with greatly improved fluorescence properties. Journal of biological chemistry, 260(6), 3440-3450.
66. Cai, Z., Ye, Z., Yang, X., Chang, Y., Wang, H., Liu, Y., & Cao, A. (2011). Encapsulated enhanced green fluorescence protein in silica nanoparticle for cellular imaging. Nanoscale, 3(5), 1974-1976.
67. Jia, B., Zhang, X., Wang, B., Chen, M., Lv, F., Wang, S., & Wang, F. (2018). Dual-modal probe based on polythiophene derivative for pre-and intraoperative mapping of lymph nodes by SPECT/optical imaging. ACS applied materials & interfaces, 10(7), 6646-6651.
68. Watanabe, K., Osaka, I., Yorozuya, S., & Akagi, K. (2012). Helically π-stacked thiophene-based copolymers with circularly polarized fluorescence: high dissymmetry factors enhanced by self-ordering in chiral nematic liquid crystal phase. Chemistry of Materials, 24(6), 1011-1024.