Near-infrared surface-enhanced Raman spectroscopy (SERS) is a powerful technique for analyzing the chemical composition within a single living cell at unprecedented resolution. However, current SERS methods employing uncontrollable colloidal metal particles or non-uniformly distributed metal particles on a substrate as SERS-active sites show relatively low reliability and reproducibility. Here, we report a highly-ordered SERS-active surface that is provided by a gold nano-dots array based on thermal evaporation of gold onto an ITO surface through a nanoporous alumina mask. This new combined technique showed a broader distribution of hot spots and a higher signal-to-noise ratio than current SERS techniques due to the highly reproducible and uniform geometrical structures over a large area. This SERS-active surface was applied as cell culture system to study living cells in situ within their culture environment without any external preparation processes. We applied this newly developed method to cell-based research to differentiate cell lines, cells at different cell cycle stages, and live/dead cells. The enhanced Raman signals achieved from each cell, which represent the changes in biochemical compositions, enabled differentiation of each state and the conditions of the cells. This SERS technique employing a tightly controlled nanostructure array can potentially be applied to single cell analysis, early cancer diagnosis and cell physiology research.

In this study, in situ electrochemical synthesis of polypyrrole nanowires with nanoporous alumina template was described. The formation of highly ordered porous alumina substrate was demonstrated with Atomic Force Microscopy (AFM) and Scanning Electron Microscopy (SEM). In addition, Fourier transform infrared analysis confirmed that polypyrrole (PP) nanowires were synthesized by direct electrochemical oxidation of pyrrole. HeLa cancer cells and HMCF normal cells were immobilized on the polypyrrole nanowires/nanoporous alumina substrates to determine the effects of the substrate on the cell morphology, adhesion and proliferation as well as the biocompatibility of the substrate. Cell adhesion and proliferation were characterized using a standard MTT assay. The effects of the polypyrrole nanowires/nanoporous alumina substrate on the cell morphology were studied by AFM. The nanoporous alumina coated with polypyrrole nanowires was found to exhibit better cell adhesion and proliferation than polystyrene petridish, aluminum foil, 1st anodized and uncoated 2nd anodized alumina substrate. This study showed the potential of the polypyrrole nanowires/nanoporous alumina substrate as biocompatibility electroactive polymer substrate for both healthy and cancer cell cultures applications.

In this study, we demonstrated a novel fabrication method of three dimensional nanoporous gold thin film (NPGF) onto gold (Au) substrate using electrochemical deposition method. Scanning electron microscope (SEM) investigation reveals the formation of highly-ordered pores, approximately 30 nm in diameter and 150 nm thick. The NPGF-modified electrode shows a linear range (0.1–40 μM) for dopamine detection in the presence of ascorbic acid. The electrochemical measurements of mixtures of dopamine, ascorbic acid, and uric acid in human serum sample for real sample applications was also investigated based on differential pulse voltammetry (DPV) technique. These high sensitivity and selectivity features of the proposed NPGF biosensor offer great promise for real sample biosensor application.

HeLa cells directly immobilized on gold-patterned silicon substrate were used to assess the biological toxicity of anticancer drugs (hydroxyurea and cyclophosphamide). Immobilization of HeLa cells was confirmed by optical microscopy, and cell growth, viability and drug-related toxicity were examined by cyclic voltammetry and potentiometric stripping analysis. The voltammetric behaviors of HeLa cells displayed a quasi-reversible pattern with the peak current exhibiting a linear relationship with cell number. The attached living cells were exposed to different concentrations of hydroxyurea and cyclophosphamide as anticancer drugs, which induced the change of cyclic voltammetry current peak. As the exposed concentration of anticancer drugs was increased, the change of current peak was increased, which indicates the decrease of cell viability. Trypan Blue dyeing was performed to confirm the results of the effect of anticancer drugs on the cell viability which was obtained from cyclic voltammetry assay. The proposed direct cell immobilization method technique can be applied to the fabrication of cell chip for diagnosis, drug detection, and on-site monitoring.

In vitro assays have generally been carried out for cytological diagnosis and for evaluation of the cytotoxic effect of chemotherapeutic agents as an alternative to animal experiments. In this study, a method for fabrication and application of a gold nanoflower array on an ITO substrate for evaluation of the effect of chemotherapeutic agents on cancer cell behavior by the surface-enhanced Raman scattering (SERS) analysis, as well as the electrochemical detection was described. Due to the increased sensitivity provided by gold nanoflower substrates, the effect of chemotherapeutic agents at low concentration level was successfully detected based on SERS technique. This substrate was found to give enhanced Raman spectra with high surface plasmon field in the near infrared (NIR) spectral range, which minimize fluorescence interference and photo-toxicity. Cyclic voltammetry (CV) was further performed for confirmation of results obtained by SERS assay and showed increased intensity of current peaks for various concentrations at low levels. The developed Au nanoflowers modified ITO substrates developed in this study could be used as a simultaneous SERS and CV substrate to determine the effects of chemotherapeutic agents on cancer cells.