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1.  Dispersion and Exposure to a Cough-Generated Aerosol in a Simulated Medical Examination Room 
Few studies have quantified the dispersion of potentially infectious bioaerosols produced by patients in the health care environment and the exposure of health care workers to these particles. Controlled studies are needed to assess the spread of bioaerosols and the efficacy of different types of respiratory personal protective equipment (PPE) in preventing airborne disease transmission. An environmental chamber was equipped to simulate a patient coughing aerosol particles into a medical examination room, and a health care worker breathing while exposed to these particles. The system has three main parts: (1) a coughing simulator that expels an aerosol-laden cough through a head form; (2) a breathing simulator with a second head form that can be fitted with respiratory PPE; and (3) aerosol particle counters to measure concentrations inside and outside the PPE and at locations throughout the room. Dispersion of aerosol particles with optical diameters from 0.3 to 7.5 μm was evaluated along with the influence of breathing rate, room ventilation, and the locations of the coughing and breathing simulators. Penetration of cough aerosol particles through nine models of surgical masks and respirators placed on the breathing simulator was measured at 32 and 85 L/min flow rates and compared with the results from a standard filter tester. Results show that cough-generated aerosol particles spread rapidly throughout the room, and that within 5 min, a worker anywhere in the room would be exposed to potentially hazardous aerosols. Aerosol exposure is highest with no personal protective equipment, followed by surgical masks, and the least exposure is seen with N95 FFRs. These differences are seen regardless of breathing rate and relative position of the coughing and breathing simulators. These results provide a better understanding of the exposure of workers to cough aerosols from patients and of the relative efficacy of different types of respiratory PPE, and they will assist investigators in providing research-based recommendations for effective respiratory protection strategies in health care settings.
PMCID: PMC4688889  PMID: 23033849
airborne particles; airborne transmission; cough; health care workers; respiratory infections/prevention; respiratory protection
2.  Detection of Infectious Influenza Virus in Cough Aerosols Generated in a Simulated Patient Examination Room 
The potential for aerosol transmission of infectious influenza virus (ie, in healthcare facilities) is controversial. We constructed a simulated patient examination room that contained coughing and breathing manikins to determine whether coughed influenza was infectious and assessed the effectiveness of an N95 respirator and surgical mask in blocking transmission.
National Institute for Occupational Safety and Health aerosol samplers collected size-fractionated aerosols for 60 minutes at the mouth of the breathing manikin, beside the mouth, and at 3 other locations in the room. Total recovered virus was quantitated by quantitative polymerase chain reaction and infectivity was determined by the viral plaque assay and an enhanced infectivity assay.
Infectious influenza was recovered in all aerosol fractions (5.0% in >4 µm aerodynamic diameter, 75.5% in 1–4 µm, and 19.5% in <1 µm; n = 5). Tightly sealing a mask to the face blocked entry of 94.5% of total virus and 94.8% of infectious virus (n = 3). A tightly sealed respirator blocked 99.8% of total virus and 99.6% of infectious virus (n = 3). A poorly fitted respirator blocked 64.5% of total virus and 66.5% of infectious virus (n = 3). A mask documented to be loosely fitting by a PortaCount fit tester, to simulate how masks are worn by healthcare workers, blocked entry of 68.5% of total virus and 56.6% of infectious virus (n = 2).
These results support a role for aerosol transmission and represent the first reported laboratory study of the efficacy of masks and respirators in blocking inhalation of influenza in aerosols. The results indicate that a poorly fitted respirator performs no better than a loosely fitting mask.
PMCID: PMC4680957  PMID: 22460981
3.  Tip-Based Nanofabrication of Arbitrary Shapes of Graphene Nanoribbons for Device Applications 
RSC advances  2015;5(46):37006-37012.
Graphene nanoribbons (GNRs) have promising applications in future nanoelectronics, chemical sensing and electrical interconnects. Although there are quite a few GNR nanofabrication methods reported, a rapid and low-cost fabrication method that is capable of fabricating arbitrary shapes of GNRs with good-quality is still in demand for using GNRs for device applications. In this paper, we present a tip-based nanofabrication method capable of fabricating arbitrary shapes of GNRs. A heated atomic force microscope (AFM) tip deposits polymer nanowires atop a CVD-grown graphene surface. The polymer nanowires serve as an etch mask to define GNRs through one step of oxygen plasma etching similar to a photoresist in conventional photolithography. Various shapes of GNRs with either linear or curvilinear features are demonstrated. The width of the GNR is around 270 nm and is determined by the width of the depositing polymer nanowire, which we estimate can be scaled down 15 nms. We characterize our TBN-fabricated GNRs using Raman spectroscopy and I-V measurements. The measured sheet resistances of our GNRs fall within the range of 1.65 kΩ/□−1 – 2.64 kΩ/□−1, in agreement with previously reported values. Furthermore, we determined the high-field breakdown current density of GNRs to be approximately 2.94×108 A/cm2. This TBN process is seamlessly compatible with existing nanofabrication processes, and is particularly suitable for fabricating GNR based electronic devices including next generation DNA sequencing technologies and beyond silicon field effect transistors.
PMCID: PMC4527583  PMID: 26257891
4.  Micro-patterning of Mammalian Cells on Suspended MEMS Resonant Sensors for Long-Term Growth Measurements 
Lab on a chip  2014;14(8):1401-1404.
MEMS resonant mass sensors can measure the mass of individual cells, though long-term growth measurements are limited by the movement of cells off the sensor area. Micro-patterning techniques are a powerful approach to control the placement of individual cells in an arrayed format. In this work we present a method for micro-patterning cells on fully suspended resonant sensors through select functionalization and passivation of the chip surface. This method combines high-resolution photolithography with a blanket transfer technique for applying photoresist to avoid damaging the sensors. Cells are constrained to the patterned collagen area on the sensor by pluronic acting as a cell adhesion blocker. This micro-patterning method enables long-term growth measurements, which is demonstrated by a measurement of the change in mass of a human breast cancer cell over 18 h.
PMCID: PMC4024477  PMID: 24535001
5.  Measuring Physical Properties of Neuronal and Glial Cells with Resonant Microsensors 
Analytical Chemistry  2014;86(10):4864-4872.
Microelectromechanical systems (MEMS) resonant sensors provide a high degree of accuracy for measuring the physical properties of chemical and biological samples. These sensors enable the investigation of cellular mass and growth, though previous sensor designs have been limited to the study of homogeneous cell populations. Population heterogeneity, as is generally encountered in primary cultures, reduces measurement yield and limits the efficacy of sensor mass measurements. This paper presents a MEMS resonant pedestal sensor array fabricated over through-wafer pores compatible with vertical flow fields to increase measurement versatility (e.g., fluidic manipulation and throughput) and allow for the measurement of heterogeneous cell populations. Overall, the improved sensor increases capture by 100% at a flow rate of 2 μL/min, as characterized through microbead experiments, while maintaining measurement accuracy. Cell mass measurements of primary mouse hippocampal neurons in vitro, in the range of 0.1–0.9 ng, demonstrate the ability to investigate neuronal mass and changes in mass over time. Using an independent measurement of cell volume, we find cell density to be approximately 1.15 g/mL.
PMCID: PMC4033632  PMID: 24734874
6.  Direct-write polymer nanolithography in ultra-high vacuum 
Polymer nanostructures were directly written onto substrates in ultra-high vacuum. The polymer ink was coated onto atomic force microscope (AFM) probes that could be heated to control the ink viscosity. Then, the ink-coated probes were placed into an ultra-high vacuum (UHV) AFM and used to write polymer nanostructures on surfaces, including surfaces cleaned in UHV. Controlling the writing speed of the tip enabled the control over the number of monolayers of the polymer ink deposited on the surface from a single to tens of monolayers, with higher writing speeds generating thinner polymer nanostructures. Deposition onto silicon oxide-terminated substrates led to polymer chains standing upright on the surface, whereas deposition onto vacuum reconstructed silicon yielded polymer chains aligned along the surface.
PMCID: PMC3304329  PMID: 22428096
additive lithography; polymer; scanning probe lithography; ultra high vacuum
7.  Control of Nanoscale Environment to Improve Stability of Immobilized Proteins on Diamond Surfaces 
Advanced functional materials  2011;21(6):1040-1050.
Immunoassays for detection of bacterial pathogens rely on the selectivity and stability of bio-recognition elements such as antibodies tethered to sensor surfaces. The search for novel surfaces that improve the stability of biomolecules and assay performance has been pursued for a long time. However, the anticipated improvements in stability have not been realized in practice under physiological conditions because the surface functionalization layers on commonly used substrates, silica and gold, are themselves unstable on time scales of days. In this paper, we show that covalent linking of antibodies to diamond surfaces leads to substantial improvements in biological activity of proteins as measured by the ability to selectively capture cells of the pathogenic bacterium Escherichia coli O157:H7 even after exposure to buffer solutions at 37 °C for extended periods of time, approaching 2 weeks. Our results from ELISA, XPS, fluorescence microscopy, and MD simulations suggest that by using highly stable surface chemistry and controlling the nanoscale organization of the antibodies on the surface, it is possible to achieve significant improvements in biological activity and stability. Our findings can be easily extended to functionalization of micro and nanodimensional sensors and structures of biomedical diagnostic and therapeutic interest.
PMCID: PMC3177702  PMID: 21949497
8.  Cadherin-Mediated Cell–Cell Contact Regulates Keratinocyte Differentiation 
Cell–extracellular matrix (ECM) and cell–cell interactions regulate keratinocyte cell fate and differentiation. In the present analysis, we examined the differentiation of primary human keratinocytes cultured on micropatterned substrates that varied the extent of cell–cell contact while maintaining constant cell–ECM areas. Bowtie-shaped micropatterned areas (75–1600 µm2) were engineered to either permit or prevent cell–cell contact for pairs of adherent keratinocytes. Cell pairs with direct cell–cell contact exhibited enhanced expression of the differentiation markers involucrin and keratin 10 compared to cells with no cell–cell contact. In contrast, available cell-spreading area, as regulated by pattern size, did not alter keratinocyte involucrin expression. Disruption of E-cadherin binding by either antibody blocking or expression of a dominant-negative receptor diminished the ability of micropattern-regulated cell–cell contact to modulate involucrin expression. These results demonstrate that cadherin-mediated cell–cell contact regulates early keratinocyte differentiation independently from changes in cell shape.
PMCID: PMC2693873  PMID: 18754040

Results 1-8 (8)