High-sensitivity in vivo THz transmission imaging of early human breast cancer in a subcutaneous xenograft mouse model.

We performed in vivo THz transmission imaging study on a subcutaneous xenograft mouse model for early human breast cancer detection. With a THz-fiber-scanning transmission imaging system, we continuously monitored the growth of human breast cancer in mice. Our in vivo study not only indicates that THz transmission imaging can distinguish cancer from the surrounding fatty tissue, but also with a high sensitivity. Our in vivo study on the subcutaneous xenograft mouse model will encourage broad and further investigations for future early cancer screening by using THz imaging system.

Cell uptake and intracellular visualization using quantum dots or nuclear localization signal-modified quantum dots with gold nanoparticles as quenchers.

Understanding and controlling the interactions between nanoscale objects and living cells is of great importance for diagnostic imaging and therapeutic applications. Quantum dots (QDs) have remarkable optical characteristics, such as uniquely feature bright, photostable, tunable and narrow fluorescence emissions, as well as broad absorption spectra. Here we report a platform of using quantum dots to investigate the cell uptake and the interactions between nanoscale objects and cells. QDs are uptaken by BHK cells easily through endocytosis. We could clearly differentiate the QDs outside the cell or inside the cell by quenching the QDs with similar sized gold nanoparticles and reduce the noise of fluorescent image. Microscopic images show that QDs are homogeneously distributed within the whole cell except the nucleus. However, unmodified QDs could not penetrate the nuclear membrane and move into the nucleus. Coupling QDs with Nuclear Localization Signal (NLS, CGGGPKKKRKVGG) can significantly enhance the translocation amount of QDs into the cell and cell nucleus. This method combined with microscopy imaging system can visualize the particle delivery routes and provide valuable information in the drug/gene delivery and tumor diagnosis.

In vivo detection of cryosurgery using multiphoton and harmonic generation microscopy.

The tissue injury and the organization of collagen during cryosurgery are poorly characterized because of the lack of appropriate methodologies. In this study, we use multimodal multiphoton microscopy to assess the change of extracellular matrix after cryotreatment of skin. The cellular matrix transformations and the intercellular interactions during the wound healing process after cryolesion for mice were measured in vivo and in real-time through the dorsal skinfold chamber (DSC). Intrinsic second-harmonic generation (SHG) signals from fibrillar collagen and two-photon excited (TPE) autofluorescence from cell were collected to investigate the cryosurgical response in vivo. The TPE and SHG signals are significantly different between normal and cryotreated mice, and correlates with the wound healing process. The results suggest that this approach may be applied in real-time to noninvasively monitor the cryosurgery process and could potentially be applied to clinical evaluation.

Multiphoton imaging and quantitative analysis of collagen production by chondrogenic human mesenchymal stem cells cultured in chitosan scaffold.

We used the combined imaging modality of multiphoton autofluorescence and second-harmonic generation microscopy to investigate the chondrogenic process of human mesenchymal stem cells cultured in chitosan scaffold. Isolated human mesenchymal stem cells seeded onto chitosan scaffold were induced to undergo chondrogenesis by addition of the transforming growth factor-β3. After continuous culturing, the engineered tissues at the same scaffold location were imaged at different time points for up to 49 days. Using the acquired images of the chondrogenic process, we quantify tissue morphogenesis by monitoring the changes in multiphoton autofluorescence and second-harmonic generation signals from the engineered tissues. We found that the extracellular matrix generation can be modeled by an exponential function during the initial growth stage and that saturation occurs between days 11 and 14. Further, the growth rate of the extracellular matrix was found to increase toward the surface of the chitosan scaffold. Our work demonstrates the use of multiphoton microscopy for performing long-term monitoring and quantification of the tissue engineering process.

The discrimination of type I and type II collagen and the label-free imaging of engineered cartilage tissue.

Using excitation polarization-resolved second harmonic generation (SHG) microscopy, we measured SHG intensity as a function of the excitation polarization angle for type I and type II collagens. We determined the second order susceptibility (χ((2))) tensor ratios of type I and II collagens at each pixel, and displayed the results as images. We found that the χ((2)) tensor ratios can be used to distinguish the two types of collagen. In particular, we obtained χ(zzz)/χ(zxx) = 1.40 ± 0.04 and χ(xzx)/χ(zxx) = 0.53 ± 0.10 for type I collagen from rat tail tendon, and χ(zzz)/χ(zxx) = 1.14 ± 0.09 and χ(xzx)/χ(zxx) = 0.29 ± 0.11 for type II collagen from rat trachea cartilage. We also applied this methodology on the label-free imaging of engineered cartilage tissue which produces type I and II collagen simultaneously. By displaying the χ((2)) tensor ratios in the image format, the variation in the χ((2)) tensor ratios can be used as a contrast mechanism for distinguishing type I and II collagens.