Controlling nonspecific protein adsorption in a plug-based microfluidic system by controlling interfacial chemistry using fluorous-phase surfactants

Control of surface chemistry and protein adsorption is important for using microfluidic devices for biochemical analysis and high-throughput screening assays. This paper describes the control of protein adsorption at the liquid-liquid interface in a plug-based microfluidic system. The microfluidic system uses multiphase flows of immiscible fluorous and aqueous fluids to form plugs, which are aqueous droplets that are completely surrounded by fluorocarbon oil and do not come into direct contact with the hydrophobic surface of the microchannel. Protein adsorption at the aqueous-fluorous interface was controlled by using surfactants that were soluble in fluorocarbon oil but insoluble in aqueous solutions. Three perfluorinated alkane surfactants capped with different functional groups were used: a carboxylic acid, an alcohol, and a triethylene glycol group that was synthesized from commercially available materials. Using complementary methods of analysis, adsorption was characterized for several proteins (bovine serum albumin (BSA) and fibrinogen), including enzymes (ribonuclease A (RNase A) and alkaline phosphatase). These complementary methods involved characterizing adsorption in microliter-sized droplets by drop tensiometry and in nanoliter plugs by fluorescence microscopy and kinetic measurements of enzyme catalysis. The oligoethylene glycol-capped surfactant prevented protein adsorption in all cases. Adsorption of proteins to the carboxylic acid-capped surfactant in nanoliter plugs could be described by using the Langmuir model and tensiometry results for microliter drops. The microfluidic system was fabricated using rapid prototyping in poly(dimethylsiloxane) (PDMS). Black PDMS microfluidic devices, fabricated by curing a suspension of charcoal in PDMS, were used to measure the changes in fluorescence intensity more sensitively. This system will be useful for microfluidic bioassays, enzymatic kinetics, and protein crystallization, because it does not require surface modification during fabrication to control surface chemistry and protein adsorption.     

Comparative investigation of the surface properties of commercial titanium dental implants. Part I: chemical composition

The surfaces of five commercially available titanium implants (Brånemark Nobel Biocare, 3i ICE, 3i OSSEOTITE, ITI-TPS, and ITI-SLA) were compared by scanning electron microscopy, X-ray photoelectron spectroscopy, time-of-flight secondary ion mass spectroscopy. All five implant types were screw-shaped and fabricated from commercially pure (cp) titanium, but their surface properties differed both as regards surface morphology and surface chemical composition. The macro- and microstructure of the implant surfaces were investigated by scanning electron microscopy. The surfaces chemical composition was determined using the surface-sensitive analytical techniques of X-ray photoelectron spectroscopy and time-of-flight secondary ion spectrometry. Surface topographies were found to reflect the type of mechanical/chemical fabrication procedures applied by the manufacturers. The titanium oxide (passive) layer thickness was similar (5–6 nm) and typical for oxide films grown at or near room temperature. A variety of elements and chemical compounds not related to the metal composition were found on some implant types. They ranged from inorganic material such as sodium chloride to specific organic compounds believed to be due to contamination during fabrication or storage. The experimental findings are believed to make a contribution to a better understanding of the interplay between industrial fabrication procedure and physico-chemical implant surface properties.     

Surface-enhanced raman detection of 2,4-dinitrotoluene impurity vapor as a marker to locate landmines

Time, cost, and casualties associated with demining efforts underscore the need for improved detection techniques. Reduction in the number of false positives by directly detecting the explosive material, rather than casing material, is desirable. The desired field sensor must, at a minimum, demonstrate reproducibility, the necessary level of sensitivity, portability, instrumental stability, and fast system response times. Ideally, vibrational spectroscopic techniques have the potential to remove false positives, since every chemical has a unique bond structure. Herein, we demonstrate the capabilities of surface-enhanced Raman spectroscopy to detect the chemical vapor signature emanating from buried TNT-based landmines. We present reproducible results obtained from blind tests controlled by the Defense Advanced Research Projects Agency (DARPA) that demonstrate vapor detection of 2,4-dinitrotoluene at concentration levels of 5 ppb or less. The results presented used acquisition times of 30 s on a fieldable system and showed that SERS can be a significant improvement over current landmine detection methods.     

Off-site ignition probability of flammable gases

A key step in the assessment of risk for installations where flammable liquids or gases are stored is the estimation of ignition probability. A review of current modelling and data confirmed that ignition probability values used in risk analyses tend to be based on extrapolation of limited incident data or, in many cases, on the judgement of those conducting the safety assessment. Existing models tend to assume that ignition probability is a function of release rate (or flammable gas cloud size) alone and they do not consider location, density or type of ignition source. An alternative mathematical framework for calculating ignition probability is outlined in which the approach used is to model the distribution of likely ignition sources and to calculate ignition probability by considering whether the flammable gas cloud will reach these sources. Data are collated on the properties of ignition sources within three generic land-use types: industrial, urban and rural. These data are then incorporated into a working model for ignition probability in a form capable of being implemented within risk analysis models. The sensitivity of the model results to assumptions made in deriving the ignition source properties is discussed and the model is compared with other available ignition probability methods.     

Specificity and sensitivity of distinctive chest radiographic and/or 67Ga images in the noninvasive diagnosis of sarcoidosis

1. Experimental chronic cerebral vasospasm induced by subarachnoid haemorrhage (SAH) in rabbits was studied in order to evaluate the efficacy of repeated i.v. infusion of Iloprost (ILO). 2. The mean diameter of the basilar artery of intact animals was calculated as 737.5 +/- 52.8 microns while this value was reduced to 237.5 +/- 22.96 microns after SAH. 3. In the ILO treated group the mean diameter of the basilar artery was significantly increased and found to be 593.75 +/- 64.0 microns. 4. No change was observed in intracranial pressure (ICP) except a slight decrease in mean arterial pressure when relatively higher doses of ILO were used. 5. These results were taken as evidence of the high therapeutic value and low side effects of ILO in the treatment of persisting cerebral vasospasm due to SAH.     

Structure and intraglyceride distribution of coriolic acid

Coriolic [(R)-13-hydroxy-cis-9,trans-11-octadecadienoic] acid (III, R=Z=H) was isolated as the methyl ester from twoCoriaria seed oils in 66 and 68% yields. The double bonds and hydroxyl group were located by periodate-permanganate oxidation before, and chromic acid oxidation after, hydrogenation of the double bonds. Alternatively the positions of the functional groups were indicated by a convenient micro-ozonolysis-gas-liquid chromatographic procedure. Determination of products from partial hydrolysis of theCoriaria oils with pancreatic lipase (EC 3.1.1.3) revealed a preference of the corioloyl group for the 1,3-positions in triglyceride molecules. The possible significance of coriolic acid as an intermediate in the biogenetic conversion of linoleic acid to conjugated trienoic acids is discussed. Presented in part at the 154th American Chemical Society meeting, Chicago, Ill., September 1967. No. Utiliz. Res. Dev. Div., ARS, USDA.     

A rapid microozonolysis-GLC procedure for locating unsaturation in olefinic acids,including trienes and tetraenes

Increased versatility has been achieved in the identification of unknown olefinic fatty acids by ozonolysis. The method has been applied to purified methyl esters containing up to four double bonds. Aldehydic fragments, obtained from esters by the Stein-Nicolaides procedure (2), were determined by GLC on two columns of different polarity. Equivalent chain lengths of each fragment on the two columns provide identification. For monoenoic esters the location of the double bond is clearly indicated by the aldehyde and aldehyde-ester fragments. Dienes are identified by the aldehyde and aldehyde-ester fragments when the original chain length of the ester is known; the dialdehyde fragment provides confirmatory evidence. Trienes and tetraenes are analyzed by interrupting the ozonolysis at various times, thereby producing unsaturated, as well as saturated, aldehydes and aldehyde-esters. Unsaturated fragments locate the central or interior double bonds. Presented at the AOCS Meeting, Chicago, October 1967. No. Utiliz. Res. Dev. Div., ARS, USDA.     

Ricinoleic acid inLinum mucronatum seed oil

Linum mucronatum seed oil contains 15% ricinoleic [(+)-12-D-hydroxy-cis-9-octadecenoic] acid, previously unknown in the Linaceae. Identification was made by IR, gas liquid chromatography, nuclear magnetic resonance, ozonolysis and mass spectrometry. Other major components of the oil are oleic acid (24%) and linoleic acid (48%). No. Market. Nutr. Res. Div., ARS, USDA.