(a) (b)
Fig. 6. 3D TPP microstructure imaged by (a) 3D TPL (Insert: 2D bright-field image) and (b) 2D TPL image of the microstructure in (a) at the base.
4. Conclusions
The biocompatible 3D microstructure of polyacrylamide with AuNRs was fabricated via the TPP processing.
To avoid AuNRs damage in the 3D microstructures, the fabrication laser wavelength (at 1.0 mW) was tuned for RB TPA, not for longitudinal plasmon resonance. Furthermore, a 5.0 mW laser power at the wavelength for the longitudinal plasmon resonance was adopted to reshape the AuNRs into AuNPs in designated positions within the 3D microstructures. Furthermore, doped AuNRs within TPL act as a contrast agent for internal visualization of the fabricated 3D microstructures. The approach described in this work can be used to create microstructures for biomedical applications.
References
1. C. R. Lambert, I. E. Kochevar, and R. W. Redmond, “Differential reactivity of upper triplet states produces wavelength-dependent two-photon photosensitization using Rose Bengal,” J. Phys. Chem. B 103,
3737-3741 (1999).
2. J. D. Pitts, P. J. Campagnola, G. A. Epling, and S. L. Goodman, “Submicron multiphoton free-form
fabrication of proteins and polymers: studies of reaction efficiencies and applications in sustained release,”
Macromolecules 33, 1514-1523 (2000).
3. P. J. Campagnola, D. M. Delguidice, G. A. Epling, K. D. Hoffacker, A. R. Howell, J. D. Pitts, and S. L.
Goodman, “3-dimensional submicron polymerization of acrylamide by multiphoton excitation of xanthene dyes,” Macromolecules 33, 1511-1513 (2000).
4. S. Kawata, H. B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697-698 (2001).
5. P. Galajda and P. Ormos, “Complex micromachines produced and driven by light,” Appl. Phys. Lett. 78, 249-251 (2001).
6. T. Tanaka, H. B. Sun, and S. Kawata, “Rapid sub-diffraction-limit laser micro/nanoprocessing in a
threshold material system,” Appl. Phys. Lett. 80, 312-314 (2002).
7. M. Miwa, S. Juodkazis, T. Kawakami, S. Matsuo, and H. Misawa, “Femtosecond two-photon stereo-lithography,” Appl. Phys. A: Mater. Sci. Process. 73, 561-566 (2001).
8. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73-76 (1990).
9. T. Watanabe, M. Akiyama, K. Totani, S. M. Kuebler, F. Stellacci, W. Wenseleers, K. Braun, S. R. Marder, and J. W. Perry, “Photoresponsive hydrogel microstructure fabricated by two-photon initiated
Polymerization,” Adv. Funct. Mater. 12, 611-614 (2002).
10. Z. B. Sun, X. Z. Dong, W. Q. Chen, S. Nakanishi, M. Duan, and S. Kawata, “Multicolor polymer
nanocomposites: in situ synthesis and fabrication of 3D microstructures,” Adv. Mater. 20, 914-919 (2008).
11. A. Marcinkevicius, S. Juodkazis, M. Watanabe, M. Miwa, S. Matsuo, and H. Misawa, “Femtosecond laser-assisted three-dimensional microfabrication in silica,” Opt. Lett. 26, 277-279 (2001).
12. P. W. Wu, W. C. Cheng, I. B. Martini, B. Dunn, B. J. Schwartz, and E. Yablonovitch, “Two-photon photographic production of three-dimensional metallic structures within a dielectric matrix,” Adv. Mater.
12, 1438-1441 (2000).
13. Y. Y. Cao, N. Takeyasu, T. Tanaka, X. M. Duan, and S. Kawata, “3D metallic nanostructure fabrication by surfactant-assisted multiphoton-induced reduction,” Small 5, 1144-1148 (2009).
14. N. Takeshima, Y. Narita, T. Nagata, and S. Tanaka, “Fabrication of photonic crystals in ZnS-doped glass,”
Opt. Lett. 30, 537-539 (2005)
15. G. Zhou and M. Gu, “Direct optical fabrication of three-dimensional photonic crystals in a high refractive index LiNbO3
16. Z. B. Sun, X. Z. Dong, S. Nzkanishi, W. Q. Chen, X. M. Duan, and S. Kawata, “Log-pile photonic crystal of CdS-polymer nanocomposites fabricated by combination of two-photon polymerization and in situ synthesis,” Appl. Phys. A 86, 427-431 (2007).
crystal,” Opt. Lett. 31, 2783-2785 (2006).
17. W. S. Kuo, C. N. Chang, Y. T. Chang, M. H. Yang, Y. H. Chien, S. J. Chen, and C. S. Yeh, “Gold nanorods in photodynamic therapy, as hyperthermia agents and in near-infrared optical imaging,” Angew. Chem. Int.
Ed. 49, 2711-2715 (2010).
18. W. S. Kuo, C. M. Wu, Z. S. Yang, S. Y. Chen, C. Y. Chen, C. C. Huang, W. M. Li, C. K. Sun, and C. S. Yeh,
“Biocompatible bacteria@Au composites for application in the photothermal destruction of cancer cells,”
Chem. Commun. 37, 4430-4432 (2008).
19. W. S. Kuo, C. N. Chang, Y. T. Chang, and C. S. Yeh, “Antimicrobial gold nanorods with dual-modality photodynamic inactivation and hyperthermia,” Chem. Commun. 32, 4853-4855 (2009)
20. J. Nappa, G. Revillod, J. P. Abid, I. Russier-Antoine, C. Jonin, E. Benichou, H. H. Giraultb, and P. F.
Brevet, “Hyper-Rayleigh scattering of gold nanorods and their relationship with linear assemblies of gold nanospheres,” Faraday Discuss. 269, 935-939 (2004).
21. A. K. Singh, D. Senapati, S. Wang, J. Griffin, A. Neely, P. Candice, K. M. Naylor, B. Varisli, J. R. Kalluri, and P. C. Ray, “Gold nanorod based selective identification of Escherichia coli bacteria using two-photon Rayleigh scattering spectroscopy,” ACS Nano 3, 1906-1912 (2009).
22. Q. Liao, C. Mu, D. S. Xu, X. C. Ai, J. N. Yao, and J. P. Zhang, “Gold nanorod array with good reproducibility for high-performance surface-enhanced Raman scattering,“ Langmuir 25, 4708-4714 (2009).
23. L. Oldenburg, M. N. Hansen, D. A. Zweifel, A. Wei, and S. A. Boppart, “Plasmon-resonant gold nanorods as backscattering albedo contrast agents for optical coherent tomography, “ Opt. Express 14, 6724-6738 (2006).
24. N. J. Durr, T. Larson, D. K. Smith, B. A. Korgel, K. Sokolov, and A. Ben-Yakar, “Two-photon
luminescence imaging of cancer cells using molecularly targeted gold nanorods,” Nano. Lett. 7, 941-945 (2007).
25. M.B. Mohamed, V. Volkov, S. Link, and M. A. El-Sayed, “The 'lightning' gold nanorods: fluorescence enhancement of over a million compared to the gold metal,” Chem. Phys. Lett. 317, 517-523 (2000).
26. C. Sonnichsen, T. Franzl, T. Wilk, G. von Plessen, J. Feldmann, O. Wilson, and P. Mulvaney, “Drastic reduction of plasmon damping in gold nanorods,” Phys. Rev. Lett. 88, 077402 (2002).
27. P. Zijlstra, J. W. M. Chon, and M. Gu, “Five-dimensional optical recording mediated by surface plasmons in gold nanorods,” Nature 459, 410-413 (2009).
28. N. R. Jana, L. Gearheart, and C. J. Murphy, “Evidence for seed-mediated nucleation in the chemical reduction of gold salts to gold nanoparticles,” Chem. Mater. 13, 2313-2322 (2001).
29. J. Perez-juste, I. Pastoriza-Santos, L. M. Liz-Marzan, and P. Mulvaney, “Gold nanorods: synthesis, characterization and applications,” Coordination Chem. Rev. 249, 1870-1901 (2005).
30. Z. Zhang and T. Yagi, “Observation of group delay dispersion as a function of the pulse width in as mode locked Ti:sapphire laser,” Appl. Phys. Lett. 63, 2993-2995 (1993).
31. L. P. Cunningham, M. P. Veilleux, and P. J. Campagnola, “Freeform multiphoton excited microfabrication for biological applications using a rapid prototyping CAD-based approach,” Opt. Express 14, 8613-8621 (2006).
32. C. Xu and W. W. Webb, “Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm,” J. Opt. Soc. Am. B 13,481-491 (1996).
33. S. Link, C. Burda, B. Nikoobakht, and M. A. El-Sayed, “Laser-induced shape changes of colloidal gold nanorods using femtosecond and nanosecond laser pulses,” J. Phys. Chem. 104, 6152-6163 (2000).
34. O. Ekici, R. K. Harrison, N. J. Durr, D. S. Eversole, M. Lee, and A. Ben-Yakar, “Thermal analysis of gold nanorods heated with femtosecond laser pulses,” J. Phys. D: Appl. Phys. 41, 185501 (2008).
35. C.-Y. Lin, K.-C. Chiu, C.-Y. Chang, S.-H. Chang, T.-F. Guo, and S.-J. Chen, “Surface plasmon-enhanced and quenched two-photon excited fluorescence,” Opt. Express 18, 12807-12817 (2010).
計畫成果自評
, “Multiphoton fabrication of freeform polymer microstructures with gold nanorods,” Optics Express, vol. 18, no. 26, pp. 27550-27559, December 2010. (Impact Factor: 3.880)S.-J.
Chen*
5 篇研討會論文
, “Gold nanorods-doped BSA microstructures via multiphoton excited photochemistry,” submitted to Optics Express (2010).
W.-S. Kuo, C.-H. Lien, K.-C. Cho, C.-Y. Chang, C.-Y. Lin, P. J. Campagnola and S.-J. Chen*
,
“Three-dimensional polymer microdevices with gold nanorods,” Proc. SPIE, vol. 7757, no. 123, August 2010.
S.-J. Chen*
K.-C. Cho, C.-Y. Chang, and
, “3D plasmonic microdevices,” Optics and Photonics in Taiwan 2010, Tainan, Taiwan, December 2010. (Invited Talk)
S.-J. Chen*
C.-H. Lien, W.-S. Kuo, K.-C. Cho, and
, “Reverse-engineering femtosecond laser microprocessing for biotissues,” Optics and Photonics in Taiwan 2010, Tainan, Taiwan, December 2010.
S.-J. Chen*
, “Gold nanorods-doped BSA microstructures via multiphoton excited photochemistry,” Optics and Photonics in Taiwan 2010, Tainan, Taiwan, December 2010.
S.-J. Chen*
與本計畫部分相關成果:
, “Fabrication of 3D plasmonic biomicrodevices,” ACP 2010, Shanghai, China, December 2010. (Invited Talk)
5篇SCI論文
R.-Y. He, C.-Y. Lin, Y.-D. Su, K.-C. Chiu, N.-S. Chang, H.-L. Wu, and S.-J. Chen*
W.-S. Kuo, C.-N. Chang, Y.-H. Chien, Y.-T. Chang,
, “Imaging live cell membranes via surface plasmon-enhanced fluorescence and phase microscopy,” Optics Express, vol. 18, no. 4, pp. 3649-3659, February 2010. (Impact Factor: 3.880)
S.-J. Chen
C.-Y. Lin
, and C.-S. Yeh, “Gold nanorods in photodynamic therapy, as hyperthermia agents, and in near-infrared optical imaging” Angewandte Chemie International Edition, vol. 49, no. 15, pp. 2711-2715, April 2010. (Impact Factor: 10.879)
, K.-C. Chiu, C.-Y. Chang, S.-H. Chang, T.-F. Guo, and S.-J. Chen*
Y.-D. Su, K.-C. Chiu, N.-S. Chang, H.-L. Wu, and
, “Surface plasmon-enhanced and quenched two-photon excited fluorescence,” Optics Express, vol. 18, no. 12, pp.
12807-12817, June 2010. (Impact Factor: 3.880)
S.-J. Chen*
V. Hovhannisyan, A. Ghazaryan, Y.-F. Chen,
, “Study of cell-biosubstrate contacts via surface plasmon polariton phase microscopy,” Optics Express, vol. 18, no. 19, pp. 20125-20135, September 2010. (Impact Factor: 3.880)
S.-J. Chen*, and C.-Y. Dong*
國際合作：
, “Photophysical mechanisms of collagen modification by 80 MHz femtosecond laser,” Optics Express, vol. 18, no. 23, pp.
24037-24047, November 2010. (Impact Factor: 3.880)
Cooperated with Prof. Paul J. Campagnola, University of Connecticut Health Center (UCHC) onto
“Integrating Plasmonic methods into Multiphoton Excitation.” The PhD student, Mr. Ruei-Yu He, got the NSC grant and to study at UCHC for one year (08/2009-06/2010).
Cooperated with Prof. Peter So, MIT onto “Widefield Multiphoton Excited Fluorescence Microscopy with Adaptive Optics.” The PhD student, Mr. Keng-Chi Cho, got the NSC grant and to study at MIT for one year (08/2009-07/2010).
國科會補助專題研究計畫成果報告自評表
請就研究內容與原計畫相符程度、達成預期目標情況、研究成果之學術或應用價 值（簡要敘述成果所代表之意義、價值、影響或進一步發展之可能性） 、是否適 合在學術期刊發表或申請專利、主要發現或其他有關價值等，作一綜合評估。
1. 請就研究內容與原計畫相符程度、達成預期目標情況作一綜合評估
■ 達成目標
□ 未達成目標（請說明，以 100 字為限）
□ 實驗失敗
□ 因故實驗中斷
□ 其他原因 說明：
2. 研究成果在學術期刊發表或申請專利等情形：
論文：■已發表 □未發表之文稿 □撰寫中 □無 專利：□已獲得 □申請中 □無
技轉：□已技轉 □洽談中 □無 其他： （以 100 字為限）
3. 請依學術成就、技術創新、社會影響等方面，評估研究成果之學術或應用價 值（簡要敘述成果所代表之意義、價值、影響或進一步發展之可能性）（以 500 字為限）
國科會補助計畫衍生研發成果推廣資料表
日期：100 年 01 月 07 日
國科會補助計畫
計畫名稱：以多光子激發於透明質酸組織膠中製造小血管與分子 影像分析
計畫主持人：陳顯禎
計畫編號：NSC 98-3111-B-006 -004 學門領域：：幹細胞/再生生 物
研發成果名稱
（中文）無
（英文）
成果歸屬機構 發明人
(創作人)
技術說明
（中文）
（200-500 字）
（英文）
產業別
技術/產品應用範圍
技術移轉可行性及預期 效益
註：本項研發成果若尚未申請專利，請勿揭露可申請專利之主要內容。
行政院國家科學委員會補助國內專家學者出席國際學術會議報告
(中文) 2010 SPIE Optics & Photonic (英文) 2010 SPIE 光電研討會 發表
論文 題目
(中文) 廣視域多光子激發螢光顯微術於活體動物之研究
(英文) Widefiled multiphoton excited florescence microcopy for animal study in vivo
報告內容：
一、參加會議經過：2010 SPIE 光電研討會：8/01-8/05
2010 SPIE 光電研討會於 2010 年 8 月 01 日至 8 月 05 日於美國加州San Diego市盛 大舉行。參加人員來自世界各國與光電工程研發相關人員，本人主要參加電漿子領域實 證運用及理論的發展(Plasmonics: Metallic Nanostructures and Their Optical Properties VIII)，以及光鑷子的操控與應用(Optical Trapping and Optical Micromanipulations VII)。
奈米電漿子近幾年的發展迅速，利用奈米電漿子晶體可突破光的繞射極限問題，達到次 波長光波導結構、超解析力之成像透鏡、超高靈敏度之感測器，因此美國重要研究經費 補助單位(如NSF或Air Force等)，投下相當大的研究經費，以期奈米電漿子能於科學研究 與工程應用上，提供嶄新的進步，解決近百年來光波繞射的限制問題。會議期間本人實 驗室有四篇壁報論文發表，除上述外，另有三篇論文：“Three-dimensional polymer microdevices with gold nanorods”, “Surface plasmon-enhanced and quenched two-photon excited fluorescence”以及 “Widefield multiphoton excited fluorescence microscopy with adaptive optics”。除與參加此次會議外，尚與三位博士生同學與一位博士後一起前往美 國加州大學聖地牙哥分校(UCSD)與洛杉磯分校(UCLA)參訪，與UCSD Prof. Zhaowei Liu 討論電漿子超解析度顯微術，UCLA Prof. Eric Chiou於Micro-optics & Micro-fluidics參 訪，以及UCLA Prof. T.-C. Tsao 與 Prof. S. Gibson 於Adaptive Optics技術合作交流。
三、考察參觀活動
於會議期間並參加 Optics & Photonics 之相關活動與展覽，不只針對光電之相關產品設備 加以分析與了解；圖一為會議之 Poster Session，與其他與會學者進行交流討論。並與三位博 士生同學與一位博士後一起前往 UCSD 及 UCLA 進行技術交流合作。
UCSD & UCLA 參訪：8/05~8/06
UCSD Prof. Zhaowei Liu 討論電漿子超解析度顯微術於生醫影像的可能性與實際應用(圖 二)，Prof. Liu 於此方面已有初步成果，其已得到初步成果，論文發表於 Science 與 Nano Letters 上。與 UCLA 機械系 Prof. S. Gibson 與 Prof. T.-C. Tsao 技術合作交流(圖三)，主要為適應性 光學的系統與理論、光學顯微技術。另外，參訪 UCLA Prof. Eric Chiou 於 Micro-optics &
Micro-fluidics 的研究(圖四)。
四、建議
世界各國利用電漿子光電技術於生物感測知應用，已有大幅進展，並陸續有商品化產品 出現，為了使我國在光學工程在生化科技應用下，能與世界主流接軌，並提高台灣生醫光電 工程之研究水準，應藉由此次的交流，可以充分了解本身和國外相關領域的差距。同時，也 了解到我們所必須加強之的地方，這個會議相當重要，每年應增加參加人數，提高台灣於國 際奈米電漿子光電之能見度。
圖一 SPIE Annual Meeting at San Diego 圖二 UCSD Visiting with Prof. Liu
圖三 UCLA Visiting with Profs. Tsao & Gibson 圖四 UCLA Visiting with Prof. Chiou
行政院國家科學委員會補助國內專家學者出席國際學術會議報告
(英文) 2010 SPIE Optics & Photonics 發表論文 題目
(中文) 雙光子激發螢光之表面電漿強化與消光之研究
(英文) Surface plasmon-enhanced and quenched two-photon excited fluorescence
報告內容：
一、參加會議經過：2010 SPIE 國際光學暨光子研討會： 7/29-8/15
SPIE(Society of Photographic Instrumentation Engineers)，其成立於 1955 年，此學會 主要係針對 “光” 有關的科學或其應用之發展與技術推進，討論的範疇包含光學、光 子學、電子學以及工程應用等發展為主。每年SPIE都會舉辦許多國際性技術研討會與短 期課程等，並具有極高學術價值。今年八月份SPIE所舉辦的Optics and Photonics國際研 討會，包含四大主題：NanoScience and Engineering, Solar Energy and Technology, Photonic Devices and Applications, and Optical Engineering and Applications。本人投稿於此研討會 並有幸獲得Poster的機會，因此前往參加，與會中不僅能和國外學者交流之下開拓自己 (Plasmonics: Metallic Nanostructures and Their Optical Properties VIII)，以及光鑷子的操控 與應用(Optical Trapping and Optical Micromanipulations VII)。在會議中可看出哪幾個研 究領域開始熱門而哪些題目開始較少團隊在研究(走下坡)，算是了解目前世界中哪些研 螢光之表面電漿強化與消光之研究(Surface plasmon-enhanced and quenched two-photon excited fluorescence)，在這個session中向各國學者介紹我們所做的研究成果，不僅與先 進們在研究上交流外對本人也具有語言訓練之目的，在這樣討論之下本人獲益良多。圖 一為本人在會場旁所拍的照片，圖二為本人在Poster session時所拍攝的照片。
附件三
除了參加此次會議外，尚與老師還有兩位博士班同學及一位博士後一起前往美國加州大學聖 地牙哥分校(UCSD)、洛杉磯分校(UCLA)參訪，結束後趁這次到美國的機會加上機票費差不 多的前提下安排前往波士頓參訪哈佛大學(Harvard)、Harvard Medical School、Massachusetts General Hospital (MGH)生物研究中心及麻省理工學院(MIT)等知名研究中心與大學。我們到 UCSD 與 Prof. Zhaowei Liu 討論電漿子超解析度顯微術，與 UCLA Prof. Eric Chiou 參訪有關 Micro-optics & Micro-fluidics 研究，UCLA Prof. T.-C. Tsao 與 Prof. S. Gibson 探討 Adaptive Optics 之技術交流，再到 Harvard 的 Medical School 拜訪 Prof. Charles Lin 參觀其實驗室的 Confocal 與 multiphoton 顯微鏡並討論藉由 Adaptive Optics 技術應用於此顯微鏡之優劣等，
並與 MIT Prof. Peter T. C. So 實驗室討論 multiphoton 顯微鏡快速取像等技術。 發螢光之表面電漿強化與消光之研究(Surface plasmon-enhanced and quenched two-photon excited fluorescence)，在這個 session 中不時都有向各國學者過來觀看也有針對內容並提問， California, San Diego, UCSD) 拜訪電子與電腦工程系(Electrical and Computer Engineering)的 Prof. Zhaowei Liu。並與 UCSD Prof. Zhaowei Liu 討論有關電漿子超解析度(superresolution)顯 微術於生醫影像的可能性與實際應用，其主要技術是靠著奈米結構產生表面電漿，其近場的 光訊號(電漿子)透過這個奈米結構(透鏡)藉由表面電漿耦合將光引至遠場，因此可以突破物理 光學的繞射極限，Prof. Liu 在此技術上發表數篇 Sicence、Nature、Nano Letters 等國際知名期 刊的文章;在談吐之中學習到要做出這樣頂尖的研究成果可是不容易，光一個結構就花費 2 年的時間沒做出來且加上龐大的奈米製程費，到第三年才有成果，由此看的出研究有時真的 必須耐得住寂寞與對事情的堅持到底。同時與他討論了許多 plasmonics 目前所面臨之相關理 論上與實驗上的問題，收穫很多。圖三係與Prof. Zhaowei Liu 合照的照片。
UCLA 參訪：8/06
2010 年 8 月 6 日參訪美國加州大學洛杉磯分校(University of California, Los Angeles,
2010 年 8 月 6 日參訪美國加州大學洛杉磯分校(University of California, Los Angeles,