In this work, we study epithelial cell growth on substrates decorated with gold nanorods that are functionalized either with a positively charged cytotoxic surfactant or with a biocompatible polymer exhibiting one of two different end groups, resulting in a neutral or negative surface charge of the particle. Upon observation of cell growth for three days by live cell imaging using optical dark field microscopy, it was found that all particles supported cell adhesion while no directed cell migration and no significant particle internalization occurred. Concerning cell adhesion and spreading as compared to cell growth on bare substrates after 3 days of incubation, a reduction by 45% and 95%, respectively, for the surfactant particle coating was observed, whereas the amino-terminated polymer induced a reduction by 30% and 40%, respectively, which is absent for the carboxy-terminated polymer. Furthermore, interface-sensitive impedance spectroscopy (electric cell-substrate impedance sensing, ECIS) was employed in order to investigate the micromotility of cells added to substrates decorated with various amounts of surfactant-coated particles. A surface density of 65 particles/µm(2) (which corresponds to 0.5% of surface coverage with nanoparticles) diminishes micromotion by 25% as compared to bare substrates after 35 hours of incubation. We conclude that the surface coating of the gold nanorods, which were applied to the basolateral side of the cells, has a recognizable influence on the growth behavior and thus the coating should be carefully selected for biomedical applications of nanoparticles.

Toxicological effects of nanoparticles are associated with their internalization into cells. Hence, there is a strong need for techniques revealing the interaction between particles and cells as well as quantifying the uptake at the same time. For that reason, herein optical dark-field microscopy is used in conjunction with transmission electron microscopy to investigate the uptake of gold nanoparticles into epithelial cells with respect to shape, stabilizing agent, and surface charge. The number of internalized particles is strongly dependent on the stabilizing agent, but not on the particle shape. A test of metabolic activity shows no direct correlation with the number of internalized particles. Therefore, particle properties besides coating and shape are suspected to contribute to the observed toxicity.

Poly(butadiene)-b-poly(ethylene oxide) vesicles were successfully loaded with hydrophilic dye Phloxine B. Dye addition during vesicle formation leads to Phloxine B encapsulated inside the water filled vesicle core as well as to freely diffusing dye molecules. The removal of uncapsulated substrate involves time consuming methods like dialysis or harsher methods like ultra filtration or selective precipitation, posing the risk of irreversible sample manipulation. Here used Phloxine B as pH sensitive fluorescence indicator allows the characterization of hydrophilic loading without separation procedure by adjusting the pH value. Additionally membrane blocking efficiency can be studied by time dependent fluorescence measurements. Cryogenic TEM studies showed that the self-assembled structure remained unchanged when the hydrophilic dye was incorporated within the vesicles. Fluorescence microscopy imaging proved the encapsulation of the hydrophilic dye inside the core volume. The combination of fluorescence correlation spectroscopy (FCS) and dynamic light scattering (DLS) measurements as ensemble methods confirmed those results additionally.

Nanoparticle exposure is monitored by a combination of two label-free and non-invasive biosensor devices which detect cellular shape and viscoelasticity (quartz crystal microbalance), cell motility and the dynamics of epithelial cell-cell contacts (electric cell-substrate impedance sensing). With these tools we have studied the impact of nanoparticle shape on cellular physiology. Gold (Au) nanoparticles coated with CTAB were synthesized and studied in two distinct shapes: Spheres with a diameter of (43 ± 4) nm and rods with a size of (38 ± 7) nm × (17 ± 3) nm. Dose-response experiments were accompanied by conventional cytotoxicity tests as well as fluorescence and dark-field microscopy to visualize the intracellular particle distribution. We found that spherical gold nanoparticles with identical surface functionalization are generally more toxic and more efficiently ingested than rod-shaped particles. We largely attribute the higher toxicity of CTAB-coated spheres as compared to rod-shaped particles to a higher release of toxic CTAB upon intracellular aggregation.

In the growing field of nanotechnology, there is an urgent need to sensitively determine the toxicity of nanoparticles since many technical and medical applications are based on controlled exposure to particles, that is, as contrast agents or for drug delivery. Before the in vivo implementation, in vitro cell experiments are required to achieve a detailed knowledge of toxicity and biodegradation as a function of the nanoparticles' physical and chemical properties. In this study, we show that the micromotility of animal cells as monitored by electrical cell-substrate impedance analysis (ECIS) is highly suitable to quantify in vitro cytotoxicity of semiconductor quantum dots and gold nanorods. The method is validated by conventional cytotoxicity testing and accompanied by fluorescence and dark-field microscopy to visualize changes in the cytoskeleton integrity and to determine the location of the particles within the cell.

The strongly polarized light scattering of gold nanorods at their longitudinal plasmon frequency allows for the tracking of single gold nanorod lateral positions and orientations via optical dark-field microscopy. We monitor both lateral and rotational diffusion of polymer-coated gold nanorods attached to artificial biomembranes on solid supports. The attachment is mediated by the biotin-streptavidin receptor-ligand system, but weak interaction is also observed in the absence of streptavidin. In the latter case, we observe a two-dimensional lateral diffusion of the nanorods with a diffusion coefficient of 0.5 mu m(2) s(-1). This lateral motion is strongly reduced with the addition of streptavidin. However, the particles are still able to rotate, and we study their rotational motion using polarization contrast microscopy. The rotational diffusion time in the range of 100 ms depends on the biotin concentration in the membrane, hence the number of anchor points, and oil the temperature. Cooling the membrane beyond its gel-fluid transition point leads to a reduction of the rotational motion. The experimental results can be understood in terms of dragging forces introduced by. the surface viscosity of the membrane. A quantitative analysis shows that entire patches of the membrane move with the particles.

We report extracellular mycosynthesis of silver nanoparticles by Fusarium acuminatum Ell. and Ev. (USM-3793) isolated from infected ginger (Zingiber officinale). An aqueous silver nitrate solution was reduced to metallic silver when exposed to F. acuminatum cell extract leading to the appearance of a brown color within 15-20 minutes. The color is due to the formation of silver nanoparticles and the excitation of surface plasmons. The optical spectrum showed the plasmon resonance at 420 nm and analysis by transmission electron microscopy confirmed the presence of silver nanoparticles. The nanoparticles produced were spherical with a broad size distribution in the range of 5-40 nm with average diameter of 13 nm. The reduction of the silver ions occurs probably by a nitrate-dependent reductase enzyme, which we found to be present in the extra-cellular medium. We tested the silver particles for their broad-band antibacterial activity on different human pathogens. We observed efficient antibacterial activity against multidrug resistant and highly pathogenic bacteria, including multidrug resistant Staphylococcus aureus, Salmonella typhi, Staphylococcus epidermidis, and Escherichia coli. The synthesis of silver nanoparticles by the fungus F. acuminatum may therefore serve as a simple, cheap, eco-friendly, reliable and safe method to produce an antimicrobial material.

We report on the successful encapsulation of hydrophobic substrates into the hydrophobic shell of poly(butadiene)-b-poly(ethylene oxide) vesicles in water and straightforward characterization methods. The hydrophobic fluorescent dye Nile Red was embedded into unilamellar vesicles of PB130-b-PEO66 prepared via different methods leading to different average hydrodynamic radii and distributions but a common hydrophobic shell thickness of similar to 16 nm as determined by transmission electron microscopy (TEM). The combination of cryogenic TEM and fluorescence microscopy studies shows that the self-assembled structure remains unchanged when the hydrophobic dye is incorporated within the vesicle shell. Furthermore, highly fluorescent quantum dots with an average diameter of d = 5.7 +/- 0.6 nm as determined by TEM were selected as hydrophobic model substrates and successfully enclosed into the vesicles, as evidenced by fluorescence correlation spectroscopy (FCS) measurements in combination with dynamic light scattering (DLS). Cryogenic TEM imaging reveals the position of the quantum dots, centered inside the double layer of the vesicle shell.

We present a general strategy to stabilize gold nanorod suspensions with mono- and bifunctional polyethylene glycol (PEG) and to attach a controlled number of nanoparticles or biomolecules. Characterization by gel electrophoresis, transmission electron microscopy (TEM), and optical dark-field microscopy show the specific binding of functionalized nanorods to their target while avoiding nonspecific binding to substrates, matrices, and other particles. Such nanorods are well suited for self-assembly of nanostructures and single-molecule labeling.

We demonstrate the separation of gold and silver nanoparticles according to their size and shape by agarose gel electrophoresis after coating them with a charged polymer layer. The separation is monitored optically using the size- and shape-dependent plasmon resonance of noble metal particles and confirmed by transmission electron microscopy (TEM). Electrophoretic mobilities are quantitatively explained by a model based on the Henry formula, providing a theoretical framework for predicting gel mobilities of polymer coated nanoparticles.