Fabrication and modification of dual-faced nano-mushrooms for tri-functional cell theranostics: SERS/fluorescence signaling, protein targeting, and drug delivery† Hsin-Yi Hsieh,‡ a Tsu-Wei Huang,‡ b Jian-Long Xiao, cd Chung-Shi Yang, be Chien-Cheng Chang, f Chin-Chou Chu, f Leu-Wei Lo, g Shenq-Hann Wang, b Pen-Cheng Wang, b Ching-Chang Chieng, b Chau-Hwang Lee * cd and Fan-Gang Tseng * abc Received 10th May 2012, Accepted 27th June 2012 DOI: 10.1039/c2jm32967k Smart nanocarriers with high SERS enhancement are inevitably important in cancer therapy for treating cells and recording their changes at the single-molecule scale. Therefore, this work reports an approach to fabricating and modifying mass-produced dual-faced and tri-functional nanoparticles for sequentially recognizing tumor cells, drug delivery, and real-time monitoring of biological responses. A one-step oxygen plasma process was employed to tailor commercially available fluorescent polystyrene beads into a corrugated upper hemisphere and simultaneously modify the entire surface with carboxylic groups. After depositing gold onto the corrugated hemisphere for SERS (using 633 nm laser excitation) while leaving the other smooth and clean hemisphere for fluorescence detection (via 488 nm or 532 nm laser excitation), the Au-coated fluorescent nano-mushrooms (AuFNMs) are formed with dual- surfaces of plasmonic gold semishells on the top and fluorescent carboxylated polystyrene at the bottom. Based on the unique structures and the self-generated functional surfaces of the AuFNMs, sulfo-NHS–SS–biotin disulfide linkers and anti-CD44 monoclonal antibodies can be simultaneously modified and added onto the top gold surfaces and the bottom carboxyl groups through Au–S and peptide bonds, respectively. The surface-modified AuFNM suspension, with >99% purity and uniform particle size in a concentration of 10 10 particles per mL in 2 mL DI water, can be employed to target overexpressive glycoproteins (CD44) on the surfaces of cancer cells and release their loads via the cleavage of disulfide bonds in the cytoplasm. These AuFNMs exhibit a 12-fold higher cancer targeting ability on HeLa cells when compared to a normal chondrocyte cell. Three-dimensional confocal particle tracking and Raman mapping are used to demonstrate the AuFNMs’ long-lasting single- particle fluorescence and superior biomolecule sensing ability. 1. Introduction Performing as miniature nanodevices for biological research, smart multifunctional nanoparticles can combine two or more characteristics, such as high image contrast, 1–3 fluorescence emission, 2,4,5 thermal therapy, 1,3 cancer targeting, 4,5 and/or drug delivery. 4,5 These characteristics allow nanoparticles to be used to simultaneously investigate complicated cell behaviors, visualize particle distribution, and/or treat cancer. In the past decade, label- free Raman spectroscopy incorporated with surface-enhanced a Institute of NanoEngineering and MicroSystems (NEMS), National Tsing Hua University, No. 101, Sec. 2, Kuang-Fu Rd., Hsinchu 30013, Taiwan R.O.C. E-mail: fangang@ess.nthu.edu.tw; Fax: +886-3- 5720724; Tel: +886-3-5715131 ext. 34270 b Department of Engineering and System Science, National Tsing Hua University, No. 101, Sec. 2, Kuang-Fu Rd., Hsinchu 30013, Taiwan R.O.C. c Research Center for Applied Sciences, Academia Sinica, No. 128, Sec. 2, Academia Rd., Nankang, Taipei 11529, Taiwan R.O.C. E-mail: clee@ gate.sinica.edu.tw; Fax: +886-2-27826680; Tel: +886-2-27898000 ext. 18 or 53 d Institute of Biophotonics, National Yang-Ming University, No.155, Sec. 2, Li-Nong St., Taipei 11221, Taiwan R.O.C. e Center for Nanomedicine Research, National Health Research Institutes, No. 35, Keyan Rd., Zhunan Town, Miaoli 35053, Taiwan R.O.C. f Institute of Applied Mechanics, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei 10617, Taiwan R.O.C. g Division of Medical Engineering Research, National Health Research Institutes, No. 35, Keyan Rd., Zhunan Town, Miaoli 35053, Taiwan R.O.C. † Electronic supplementary information (ESI) available: Monodisperse AuFNM treatment processes, PSB spin-coating results, SEM and/or TEM images of oxygen plasma etched PSBs, AuFNMs, and/or QD modified AuFNMs, UV-vis spectrum of AuFNMs, multifunction illustration of surface modified AuFNMs, and other supplementary movies for particle visualization and confocal reconstruction images. See DOI: 10.1039/c2jm32967k ‡ These two authors have made equal contributions to this paper. 20918 | J. Mater. Chem., 2012, 22, 20918–20928 This journal is ª The Royal Society of Chemistry 2012 Dynamic Article Links C < Journal of Materials Chemistry Cite this: J. Mater. Chem., 2012, 22, 20918 www.rsc.org/materials PAPER Downloaded by National Tsing Hua University on 14 January 2013 Published on 06 August 2012 on http://pubs.rsc.org | doi:10.1039/C2JM32967K View Article Online / Journal Homepage / Table of Contents for this issue