We demonstrated that lipid-enclosed CdSe quantum dots (LEQDs) can function as versatile contrast agents in epi-detection third harmonic generation (THG) microscopy for biological applications in vivo. With epi-THG intensities 20 times stronger than corresponding fluorescence intensities from the same LEQDs under the same conditions of energy absorption, such high brightness LEQDs were proved for the abilities of cell tracking and detection of specific molecular expression in live cancer cells. Using nude mice as an animal model, the distribution of LEQD-loaded tumor cells deep in subcutaneous tissues were imaged with high THG contrast. This is the first demonstration that THG contrast can be manipulated in vivo with nanoparticles. By linking LEQDs with anti-Her2 antibodies, the expression of Her2/neu receptors in live breast cancer cells could also be easily detected through THG. Compared with fluorescence modalities, the THG modality also provides the advantage of no photobleaching and photoblinking effects. Combined with a high penetration 1230 nm laser, these novel features make LEQDs excellent THG contrast agents for in vivo deep-tissue imaging in the future.

© 2008 Optical Society of America

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Adv. Mater.

• S.-P. Tai, Y. Wu, D.-B. Shieh, L.-J. Chen, K.-J. Lin, C.-H. Yu, S.-W. Chu, C.-H. Chang, X.-Y. Shi, Y.-C. Wen, K.-H. Lin, T.-M. Liu, and C.-K. Sun, "Molecular Imaging of Cancer Cells Using Plasmon-Resonant-Enhanced Third-Harmonic-Generation in Silver Nanoparticles," Adv. Mater. 19, 4520-4523 (2007). [CrossRef]

Anal. Chem

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Appl. Phys. Lett.

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Br. J. Cancer

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• D. Mohanta, G. A. Ahmed, A. Choudhury, F. Singh, D. K. Avasthi, G. Boyer, and G. A. Stanciu, "Scanning probe microscopy, luminescence and third harmonic generation studies of elongated CdS:Mn nanostructures developed by energetic oxygen-ion-impact," Eur. Phys. J.: Appl. Phys. 35, 29-36 (2006). [CrossRef]

J. Am. Chem. Soc.

• C. B. Murray, D. J. Norris, and M. G. Bawendi, "Synthesis and characterization of nearly monodisperse CdE (E= sulfur, selenium, tellurium) semiconductor nanocrystallites," J. Am. Chem. Soc. 115, 8706-8715 (1993). [CrossRef]

J. Biomed. Opt.

• S.-Y. Chen, C.-S. Hsieh, S.-W. Chu, C.-Y. Lin, C.-Y. Ko, Y.-C. Chen, H.-J. Tsai, C.-H. Hu, and C.-K. Sun, "Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo," J. Biomed. Opt. 11, 054022 (2006). [CrossRef] [PubMed]

J. Cell. Physiol.

• M. Campiglio, A. Locatelli, C. Olgiati, N. Normanno, G. Somenzi, L. Vigano, M. Fumagalli, S. Menard, and L. Gianni, "Inhibition of proliferation and induction of apoptosis in breast cancer cells by the epidermal growth factor receptor(EGFR) tyrosine kinase inhibitor ZD 1839(???Iressa???) is independent of EGFR expression level," J. Cell. Physiol. 198, 259-268 (2004). [CrossRef]

J. Clin. Oncol.

• O. Thuerigen, A. Schneeweiss, G. Toedt, P. Warnat, M. Halm, H. Kramer, B. Brors, C. Rudlowski, A. Benner, F. Schuetz, B. Tews, R. Eils, H. P. Sinn, C. Sohn, and P. Lichter, "Gene expression signature predicting pathologic complete response with gemcitabine, epirubicin, and docetaxel in primary breast cancer," J. Clin. Oncol. 24, 1839-1845 (2006). [CrossRef] [PubMed]

J. Invest. Dermatol.

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J. Microsc.

• E. J. Gualda, G. Filippidis, G. Voglis, M. Mari, C. Fotakis, and N. Tavernarakis, "In vivo imaging of cellular structures in Caenorhabditis elegans by combined TPEF, SHG and THG microscopy," J. Microsc. 229, 141-150 (2008). [CrossRef] [PubMed]
• S.-W. Chu, I.-H. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002). [CrossRef] [PubMed]

J. Opt. Soc. Am. B: Opt. Phys.

• G. O. Clay, A. C. Millard, C. B. Schaffer, J. Aus-der-Au, P. S. Tsai, J. A. Squier, and D. Kleinfeld, "Spectroscopy of third-harmonic generation: evidence for resonances in model compounds and ligated hemoglobin," J. Opt. Soc. Am. B: Opt. Phys. 23, 932-950 (2006). [CrossRef]
• J. M. Schins, T. Schrama, J. Squier, G. J. Brakenhoff, and M. Muller, "Determination of material properties by use of third-harmonic generation microscopy," J. Opt. Soc. Am. B: Opt. Phys. 19, 1627-1634 (2002). [CrossRef]
• J. X. Cheng and X. S. Xie, "Green's function formulation for third-harmonic generation microscopy," J. Opt. Soc. Am. B: Opt. Phys. 19, 1604-1610 (2002). [CrossRef]

J. Phys. Chem.

• M. A. Hines and P. Guyot-Sionnest, "Synthesis and characterization of strongly luminescing ZnS-Capped CdSe nanocrystals," J. Phys. Chem. 100, 468-471 (1996). [CrossRef]

J. Phys.: Condens. Matter

• A. K. Dharmadhikari, N. Kumbhojkar, J. A. Dharmadhikari, S. Mahamuni, and R. C. Aiyer, "Studies on third-harmonic generation in chemically grown ZnS quantum dots," J. Phys.: Condens. Matter 11, 1363-1368 (1999). [CrossRef]

J. Phys.: Conf. Ser.

• J. T. Seo, S. M. Ma, Q. Yang, L. Creekmore, R. Battle, H. Brown, A. Jackson, T. Skyles, B. Tabibi, and W. Yu, "Large Resonant Third-order Optical Nonlinearity of CdSe Nanocrystal Quantum Dots," J. Phys.: Conf. Ser. 38, 91-94 (2006). [CrossRef]

J. Struct. Biol.

• C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004). [CrossRef] [PubMed]

Microsc. Res. Tech

• S.-W. Chu, S.-P. Tai, C.-L. Ho, C.-H. Lin, and C.-K. Sun, "High-resolution simultaneous three-photon fluorescence and third-harmonic-generation microscopy," Microsc. Res. Tech 66, 193-197 (2005). [CrossRef] [PubMed]

Mol. Carcinog.

• S. S. Bacus, K. Kiguchi, D. Chin, C. R. King, and E. Huberman, "Differentiation of Cultured Human Breast Cancer Cells (AU-565 and MCF-7) Associated with Loss of Cell Surface HER-2/neu Antigen," Mol. Carcinog. 3, 350-362 (1990). [CrossRef] [PubMed]

N. Engl. J. Med.

• K. I. Pritchard, L. E. Shepherd, F. P. O'Malley, I. L. Andrulis, D. S. Tu, V. H. Bramwell, and M. N. Levine, "HER2 and responsiveness of breast cancer to adjuvant chemotherapy," N. Engl. J. Med. 354, 2103-2111 (2006). [CrossRef] [PubMed]

Nano Lett.

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