Michael D. Mason

PEGylated Gold Nanoparticles Conjugated to Monoclonal F19 Antibodies as Targeted Labeling Agents for Human Pancreatic Carcinoma Tissue

In this study, we describe optical detection of antibody-conjugated nanoparticles bound to surgically resected human pancreatic cancer tissue. Gold nanoparticles stabilized by heterobifunctional polyethylene glycol (PEG) were prepared using approximately 15 nm spherical gold cores and covalently coupled to F19 monoclonal antibodies. The heterobifunctional PEG ligands contain a dithiol group for stable anchoring onto the gold surface and a terminal carboxy group for coupling of antibodies to the outside of the PEG shell. The nanoparticle-antibody bioconjugates form highly stable dispersions and exhibit long-term resistance to agglomeration. This has been demonstrated by dynamic light scattering, size exclusion chromatography, and transmission electron microscopy. The nanoparticle bioconjugates were used to label tumor stroma in approximately 5 mum thick sections of resected human pancreatic adenocarcinoma. After rinsing away nonbound nanoparticles and fixation, the tissue samples were imaged by darkfield microscopy near the nanoparticle resonance scattering maximum (approximately 560 nm). The images display pronounced tissue features and suggest that this novel labeling method could provide for facile identification of cancer tissue. Tumor samples treated with gold nanoparticles conjugated to nonspecific control antibodies and noncancerous pancreatic tissue treated with mAb-F19-conjugated gold nanoparticles both exhibited correctly negative results and showed no tissue staining.

Silver nanospheres are cytotoxic and genotoxic to fish cells

Nanoparticles are being widely investigated for a range of applications due to their unique physical properties. For example, silver nanoparticles are used in commercial products for their antibacterial and antifungal properties. Some of these products are likely to result in silver nanoparticles reaching the aquatic environment. As such, nanoparticles pose a health concern for humans and aquatic species. We used a medaka (Oryzias latipes) cell line to investigate the cytotoxicity and genotoxicity of 30nm diameter silver nanospheres. Treatments of 0.05, 0.3, 0.5, 3 and 5microg/cm(2) induced 80, 45.7, 24.3, 1 and 0.1% survival, respectively, in a colony forming assay. Silver nanoparticles also induced chromosomal aberrations and aneuploidy. Treatments of 0, 0.05, 0.1 and 0.3microg/cm(2) induced damage in 8, 10.8, 16 and 15.8% of metaphases and 10.8, 15.6, 24 and 24 total aberrations in 100 metaphases, respectively. These data show that silver nanoparticles are cytotoxic and genotoxic to fish cells.

EXTERNAL QUANTUM EFFICIENCY OF SINGLE POROUS SILICON NANOPARTICLES

We use a combination of single nanoparticle luminescence and scanning force microscopy to determine the quantum efficiency (QE) of single porous Si nanoparticles and to determine the ratio of luminescent nanoparticles deposited on a silica surface to the total nanoparticles. An estimate of the QE of bulk porous Si based on these data compares favorably to the QE measured experimentally. From this we conclude that the 1% QE of bulk porous Si measured experimentally results primarily from a statistical distribution of high QE quantum-confined Si chromophores.

Spatially resolved photoluminescence of inversion domain boundaries in GaN-based lateral polarity heterostructures

Intentionally grown GaN inversion domain boundaries (IDBs) of lateral polarity heterostructures have been spectroscopically imaged at low temperature using high spatial resolution photoluminescence. It is shown that the IDBs are not only optically active, but are more than an order of magnitude brighter than the GaN bulk material. Our findings are in agreement with calculations predicting that IDBs should not adversely affect near-band-gap photoluminescence due to the absence of midgap electronic states. Typical linewidths are on the order of 10–20 meV, however, features less than 0.6 meV are observed. The boundary emission is found to be neither spectrally nor spatially uniform. Also, a strong polarization dependence of the IDB photoluminescence is measured and determined to be oriented parallel to the boundary between GaN of N- or Ga-face polarity.

Optical Signal Comparison of Single Fluorescent Molecules and Raman Active Gold Nanostars

The relevant photophysical properties of single fluorescent molecules and single SERS active surface-coated gold nanostars tagged with the Raman reporter molecule 4-mercaptopyridine are compared for imaging purposes. Mean count rate distributions are built from the single molecule/single probe level. The individually observed variance and count rates of both systems are compared as well as the behavior over multiple image acquisitions.

Investigation of the Emission Mechanism in Milled SrAl2O4:Eu, Dy Using Optical and Synchrotron X-ray Spectroscopy

There currently exists much debate as to the active state related to the long afterglow effect in europium doped oxide materials. Redox couples that consist of Eu(+/2+) and Eu(2+/3+) are discussed, but no common answer is currently accepted. Here, we present a comparison of the optical properties of a commercially available SrAl(2)O(4):Eu, Dy phosphor, as a function of nanoparticle size reduction via dry mechanical milling. X-ray and optical spectroscopic data indicate a significant decrease in phosphorescence efficiency and an increase in laser stimulated emission efficiency as near surface Eu(2+) ions are oxidized to Eu(3+) as a consequence of increased exposure during the milling process. These results show evidence only for Eu(2+/3+) oxidation states, suggesting the mechanism related to long afterglow effect does not arise from Eu(+) species. We also suggest that size reduction, as a rule, cannot be universally applied to improve optical properties of nanostructures.

Ultrahigh resolution imaging of biomolecules by fluorescence photoactivation localization microscopy.

Diffraction limits the biological structures that can be imaged by normal light microscopy. However, recently developed techniques are breaking the limits that diffraction poses and allowing imaging of biological samples at the molecular length scale. Fluorescence photoactivation localization microscopy (FPALM) and related methods can now image molecular distributions in fixed and living cells with measured resolution better than 30 nm. Based on localization of single photoactivatable molecules, FPALM uses repeated cycles of activation, localization, and photobleaching, combined with high-sensitivity fluorescence imaging, to identify and localize large numbers of molecules within a sample. Procedures and pitfalls for construction and use of such a microscope are discussed in detail. Representative images of cytosolic proteins, membrane proteins, and other structures, as well as examples of results during acquisition are shown. It is hoped that these details can be used to perform FPALM on a variety of biological samples, to significantly advance the understanding of biological systems.

Gold Nanoparticles Provide Bright Long-Lasting Vascular Contrast for CT Imaging

OBJECTIVEnIodinated contrast agent for CT has a short half-life in the vasculature. As the field of interventional procedures expands, a more durable contrast agent would be highly useful. Our study investigated whether gold nanoparticles are feasible as a long-lasting vascular contrast agent for CT.nnnMATERIALS AND METHODSnGold nanoparticles were synthesized by a modified Turkevich method, coated with methoxy-polyethylene glycol-thiol chains, and compared with an iodine-based contrast agent used in mice. Contrast agents were imaged in tubes by CT at 40, 60, and 140 kVp and then were tested in vivo by tail vein injection. Nine mice received gold nanoparticles, two received iodine-based contrast agent, and one received saline. CT of mice was performed at 60 kVp immediately, 6 hours, and 24 hours after injection.nnnRESULTSnIn an isolated form in tubes, gold nanoparticles had greater radiographic density than did iodine-based contrast agent at 40 kVp and were comparable at the other CT voltages. In mice, gold nanoparticles provided bright contrast enhancement that enabled clear visualization of the abdominal aorta and renal arteries, which could not be distinguished without contrast agent. This persisted up to 24 hours, which was the last time point assessed. Contrast enhancement of the vasculature by iodine-based contrast agent was present immediately after injection but had disappeared by 6 hours.nnnCONCLUSIONnGold nanoparticles can provide clear and durable contrast enhancement of the vasculature even at 24 hours. These findings merit further study of gold nanoparticles for their potential as a contrast agent in CT and CT-guided interventional procedures.

The cytotoxicity and genotoxicity of soluble and particulate cobalt in human lung fibroblast cells

Cobalt exposure is increasing as cobalt demand rises worldwide due to its use in enhancing rechargeable battery efficiency, super-alloys, and magnetic products. Cobalt is considered a possible human carcinogen with the lung being a primary target. However, few studies have considered cobalt-induced toxicity in human lung cells. Therefore, in this study, we sought to determine the cytotoxicity and genotoxicity of particulate and soluble cobalt in human lung cells. Cobalt oxide and cobalt chloride were used as representative particulate and soluble cobalt compounds, respectively. Exposure to both particulate and soluble cobalt induced a concentration-dependent increase in cytotoxicity, genotoxicity, and intracellular cobalt ion levels. Based on intracellular cobalt ion levels, we found that soluble cobalt was more cytotoxic than particulate cobalt while particulate and soluble cobalt induced similar levels of genotoxicity. However, soluble cobalt induced cell cycle arrest indicated by the lack of metaphases at much lower intracellular cobalt concentrations compared to cobalt oxide. Accordingly, we investigated the role of particle internalization in cobalt oxide-induced toxicity and found that particle-cell contact was necessary to induce cytotoxicity and genotoxicity after cobalt exposure. These data indicate that cobalt compounds are cytotoxic and genotoxic to human lung fibroblasts, and solubility plays a key role in cobalt-induced lung toxicity.

Correlation between bulk morphology and luminescence in porous silicon investigated by pore collapse resulting from drying

Abstract We used a combination of scanning electron microscopy, laser scanning confocal microscopy and luminescence spectroscopy to correlate the emission properties of anodized porous silicon (PS) with film morphology in samples that have undergone solvent evaporation-induced collapse of the underlying porous structure. Several PS samples were investigated as a function of the current density (J) and total etch time, while the total charge (Q) injected per unit area (with the total amount of Si removed) was kept constant during etching. From these data, two classes of PS samples emerge. Porous silicon samples produced at high current density have a three-dimensional pore network with a narrow distribution of blue–green emitting chromophores. In contrast, low current density samples form a two-dimensional pore network normal to the Si substrate with larger chromophores and exhibit broad red luminescence.