Internalization of Auger electron-emitting isotopes into cancer cells: a method for spatial distribution determination of equivalent source terms.

PURPOSE: A challenge for single-cell dosimetry of internalized Auger electron-emitting (AE) radiopharmaceuticals remains how best to elucidate their spatial distribution. To this end, a method, photoresist autoradiography (PAR), was previously developed to identify the lateral spatial distribution of AE-emitting radionuclides internalized in single cancer cells. In this paper, we present a simple mathematical model based on the radius and depth of radiation-induced patterns in photoresist material to identify the location in the z-plane of an 111In source capable of generating the pattern. MATERIALS AND METHODS: SQ20B cells, derived from a head and neck squamous cell carcinoma, were exposed to 111In-labeled epidermal growth factor (EGF) (8 MBq/µg). The integrated electron fluence after four half-lives from the internalized radionuclide-containing construct was detected by a photoresist layer that was placed in close proximity to the cells. The resultant latent patterns were chemically developed and analyzed by atomic force microscopy (AFM). The features in the patterns were matched to locations of electrons emitted from simulated point sources, thereby determining the likely locations of internalized radionuclides. RESULTS: The modeling procedure was validated using simple patterns. The model relates the depth and radius (in the x-y plane) of a pattern to the location and fluence of the source giving rise to the pattern. This point source modeling method provided a good fit to experimental data and can be expanded to analyze more complex patterns. CONCLUSIONS: We have demonstrated the utility of the modelling technique to identify the location of internalized AE-emitting radionuclides. This methodology now needs to be extended to predict the source positions in more complex PAR patterns.

Removal mechanisms of dew via self-propulsion off the gecko skin.

Condensation resulting in the formation of water films or droplets is an unavoidable process on the cuticle or skin of many organisms. This process generally occurs under humid conditions when the temperature drops below the dew point. In this study, we have investigated dew conditions on the skin of the gecko Lucasium steindachneri. When condensation occurs, we show that small dew drops, as opposed to a thin film, form on the lizard's scales. As the droplets grow in size and merge, they can undergo self-propulsion off the skin and in the process can be carried away a sufficient distance to freely engage with external forces. We show that factors such as gravity, wind and fog provide mechanisms to remove these small droplets off the gecko skin surface. The formation of small droplets and subsequent removal from the skin may aid in reducing microbial contact (e.g. bacteria, fungi) and limit conducive growth conditions under humid environments. As well as providing an inhospitable microclimate for microorganisms, the formation and removal of small droplets may also potentially aid in other areas such as reduction and cleaning of some surface contaminants consisting of single or multiple aggregates of particles.

A gecko skin micro/nano structure - A low adhesion, superhydrophobic, anti-wetting, self-cleaning, biocompatible, antibacterial surface.

Geckos, and specifically their feet, have attracted significant attention in recent times with the focus centred around their remarkable adhesional properties. Little attention however has been dedicated to the other remaining regions of the lizard body. In this paper we present preliminary investigations into a number of notable interfacial properties of the gecko skin focusing on solid and aqueous interactions. We show that the skin of the box-patterned gecko (Lucasium sp.) consists of dome shaped scales arranged in a hexagonal patterning. The scales comprise of spinules (hairs), from several hundred nanometres to several microns in length, with a sub-micron spacing and a small radius of curvature typically from 10 to 20 nm. This micro and nano structure of the skin exhibited ultralow adhesion with contaminating particles. The topography also provides a superhydrophobic, anti-wetting barrier which can self clean by the action of low velocity rolling or impacting droplets of various size ranges from microns to several millimetres. Water droplets which are sufficiently small (10-100 µm) can easily access valleys between the scales for efficient self-cleaning and due to their dimensions can self-propel off the surface enhancing their mobility and cleaning effect. In addition, we demonstrate that the gecko skin has an antibacterial action where Gram-negative bacteria (Porphyromonas gingivalis) are killed when exposed to the surface however eukaryotic cell compatibility (with human stem cells) is demonstrated. The multifunctional features of the gecko skin provide a potential natural template for man-made applications where specific control of liquid, solid and biological contacts is required.

Characterization of single α-tracks by photoresist detection and AFM analysis-focus on biomedical science and technology.

The interactions between energetic ions and biological and/or organic target materials have recently attracted theoretical and experimental attention, due to their implications for detector and device technologies, and for therapeutic applications. Most of the attention has focused on detection of the primary ionization tracks, and their effects, while recoil target atom tracks remain largely unexplored. Detection of tracks by a negative tone photoresist (SU-8), followed by standard development, in combination with analysis by atomic force microscopy, shows that both primary and recoil tracks are revealed as conical spikes, and can be characterized at high spatial resolution. The methodology has the potential to provide detailed information about single impact events, which may lead to more effective and informative detector technologies and advanced therapeutic procedures. In comparison with current characterization methods the advantageous features include: greater spatial resolution by an order of magnitude (20 nm); detection of single primary and associated recoil tracks; increased range of fluence (to 2.5 × 10(9) cm(-2)); sensitivity to impacts at grazing angle incidence; and better definition of the lateral interaction volume in target materials.

Nanographene oxide-based radioimmunoconstructs for in vivo targeting and SPECT imaging of HER2-positive tumors.

Nanographene oxide (NGO) is a novel nano-wall material that tracks to tumors in vivo, and which, as a consequence of its large surface area, has the capacity to carry a large payload. This study explores the use of anti-HER2 antibody (trastuzumab)-conjugated NGO, radiolabeled with (111)In-benzyl-diethylenetriaminepentaacetic acid (BnDTPA) via ππ-stacking, for functional imaging. In two HER2-overexpressing murine models of human breast cancer, high tumor-to-muscle ratio was achieved, resulting in clear visualization of tumor using single-photon emission computed tomography (SPECT). In the BALB/neuT model and in BALB/c nu/nu mice bearing 231/H2N xenografts, tumor accumulation amounted to 12.7 ± 0.67 and 15.0 ± 3.7% of the injected dose/g (%ID/g) of tumor tissue at 72 h, with tumor-to-muscle ratios of 35:1 and 7:1, respectively. Radiolabeled NGO-trastuzumab conjugates demonstrated superior pharmacokinetics compared to radiolabeled trastuzumab without NGO, with more rapid clearance from the circulation. The use of NGO as a scaffold to build radiolabeled nano-immunoconstructs holds promise for molecular imaging of tumors.

Photoresists as a high spatial resolution autoradiography substrate for quantitative mapping of intra- and sub-cellular distribution of Auger electron emitting radionuclides.

PURPOSE: To explore poly(methyl methacrylate) (PMMA950) as an autoradiography substrate. MATERIALS AND METHODS: PMMA950 was spin coated onto a silicon substrate. Resists were exposed to either a 25 or 50 keV electron beam (e-beam) with fluences of 0.1-33.6 µC/cm(2). The resulting patterns were analyzed by atomic force microscopy (AFM). The dependence of pattern sensitivity and resolution on resist thickness, development time and electron energy was evaluated and correlated with Monte Carlo (MC) modeling. Conventional micro-autoradiography (MAR) images were compared to AFM images of photoresist patterns obtained following exposure from (111)In-diethylenetriaminepentaacetic acid (DTPA)-human epidermal growth factor (hEGF) (4-6 MBq/µg, 40 nM DTPA-hEGF)-treated human breast cancer cells MDA-MB-468. RESULTS: MC simulation results confirmed the similarity of particle transport in PMMA950 exposed to either an (111)In point source or a 25 keV e-beam. Sensitivity was inversely related to resist thickness. Development conditions of the resists greatly affected image quality. Sensitivity of PMMA950 was similar to the UVIII™ resist (consisting of a copolymer of 4-hydroxystyrene and t- butylacrylate) at low electron fluence for both 25 and 50 keV e-beam exposure. AFM evaluation of the exposure patterns from (111)In-DTPA-hEGF treated cells and nuclei provides more detailed information in comparison with that from MAR. CONCLUSIONS: Photoresist autoradiography can provide information on both the distribution of radiation sources and their strengths within a biological sample; however, the choice of photoresist material and processing conditions greatly affects the outcome.

Chemically amplified photoresist for high resolution autoradiography in targeted radiotherapy.

Evaluation of the intracellular distribution of radionuclides used for targeted radiotherapy (tRT) is essential for accurate dosimetry. Therefore, a direct and quantitative method for subcellular micro-autoradiography using radiation sensitive polymers (PMMA, UV1116 and AZ40XT) was developed. The electron exposure dose in radio-labelled cells due to Auger and internal conversion (IC) electron emissions of indium (¹¹¹In), a radionuclide currently used for tRT, was calculated using Monte Carlo (MC) simulation. Electron beam lithography using pre-defined exposure doses was used to calibrate the resist response. The topography of the exposed and developed resists was analysed with atomic force microscopy (AFM) and the resulting pattern depth was related to a specific exposure dose. UV1116 exhibited the best contrast as compared to AZ40XT and PMMA, while AZ40XT exhibited the highest sensitivity at low doses (<10 µC/cm²). AFM analysis of the exposure pattern from radio-labelled cells and nuclei in UV1116 revealed a non-uniform distribution of ¹¹¹In-EGF in the cell and nucleus, consistent with less well-resolved data from confocal microscopy and micro-autoradiography.

Measuring anisotropic friction on WTe2 using atomic force microscopy in the force-distance and friction modes.

Layered materials which can be easily cleaved have proved to be excellent samples for the study of atomic scale friction. The layered transition metal dichalcogenides have been particularly popular. These materials exhibit a number of interesting properties ranging from superconductivity to low frictional coefficients. In this paper we have investigated the tribology of the dichalcogenide-WTe2. The coefficient of friction is less than 0.040 along the Te rows and increases to over 0.045 across the rows. The frictional forces almost doubled at normal loads of 5000 nN when scanning in the [010] direction in comparison to the [100] direction. The frictional responses of the AFM probe have been monitored in the frictional force and force-versus-distance (f-d) mode. A comparison between the outcomes using the two different modes demonstrates the factors which need to be considered for accurate measurements.

Putative functions and functional efficiency of ordered cuticular nanoarrays on insect wings.

The putative functions and functional efficiencies of periodic nanostructures on the surface of cicada wings have been investigated by atomic force microscopy (AFM) used as a tool for imaging, manipulation, and probing of adhesion. The structures consist of hexagonal close-packed protrusions with a lateral spacing of approximately 200 nm and may have multiple functionalities. Not only do the structures confer survival value by virtue of camouflage, but they may also serve as antiwetting and self-cleaning surfaces and thus be resistant to contamination. These effects have been demonstrated by exposure to white light, liquid droplets, and AFM adhesion measurements. The dependence of optical reflectivity and surface adhesion on surface topography has been demonstrated using AFM as a nanomachining tool as well as an imaging and force-sensing probe. The intact arrays display exceptionally low adhesion for particles in the size range 20 nm-40 microm. The particles can be removed from the array by forces in the range 2-20 nN; conversely, forces in the range 25-230 nN are required to remove identical particles from a flat hydrophilic surface (i.e., polished Si). Measurements of contact angles for several liquids and particle adhesion studies show that the wing represents a low-surface-energy membrane with antiwetting properties. The inference is that a combination of chemistry and structure constitutes a natural technology for conferring resistance to contamination.

A review of enabling technologies based on scanning probe microscopy relevant to bioanalysis.

The scanning probe microscope (SPM) system is emerging as an increasingly important tool for non-intrusive interrogation of biomolecular systems in vitro. Its particular merit is that it retains complete functionality in a biocompatible fluid environment and can track the dynamics of cellular and molecular processes in real time and real space at nm resolution, as an imaging tool, and with pN force-sensing/imposing resolution, as an interaction tool. The capability may have relevance as a test bed for monitoring cellular response to environmental stimuli and pharmaceutical intervention. We shall also review the better-known recent contributions of SPM towards explanatory and predictive descriptions of biomolecular interactions at surfaces and interfaces, and describe some recent attempts to reconfigure the SPM platform for demonstration of novel bio-device applications.

Calibration of AFM cantilever spring constants.

In this paper we present two simple, reliable and readily applicable methods for calibrating cantilevers and measuring the thickness of thin gold films. The spring constant calibration requires knowledge of the Young's modulus, density of the cantilever and resonant frequency. The thickness of thin gold layers was determined by measuring changes in the resonant frequency and Q-factor of beam shaped AFM cantilevers before and after coating. The techniques for measuring the spring constant and thin film thickness provide accuracy on the order of 10-15%.