Flow reproducibility of whole blood and other bodily fluids in simplified no reaction lateral flow assay devices.

The "no reaction" lateral flow assay (nrLFA) uses a simplified LFA structure with no conjugate pad and no stored reagents. In the nrLFA, the capillary-based transport time or distance is the key indicator, rather than the outcome of a biochemical reaction. Hence, the calibration and reproducibility of the nrLFA device are critical. The capillary flow properties of several membrane types (nitrocellulose, nylon, cellulose acetate, polyethersulfone, and polyvinylidene difluoride) are evaluated. Flow rate evaluations of MilliporeSigma Hi-Flow™ Plus (HF075, HF135 and HF180) nitrocellulose membranes on nrLFA are performed using bodily fluids (whole blood, blood plasma, and artificial sweat). The results demonstrate that fluids with lower viscosity travel faster, and membranes with slower flow rate exhibit higher capability to distinguish fluids with different viscosities. Reproducibility tests of nrLFA are performed on HF075, demonstrating excellent reproducibility. The coefficient of variation for blood coagulation tests performed with the nrLFA using induced coagulation was 5% for the plasma front and 2% for the RBC front. The effects of variation in blood hematocrit and sample volume are also reported. The overall results indicate that the nrLFA approach has a high potential to be commercially developed as a blood monitoring point-of-care device with simple calibration capability and excellent reproducibility.

Blood coagulation screening using a paper-based microfluidic lateral flow device.

A simple approach to the evaluation of blood coagulation using a microfluidic paper-based lateral flow assay (LFA) device for point-of-care (POC) and self-monitoring screening is reported. The device utilizes whole blood, without the need for prior separation of plasma from red blood cells (RBC). Experiments were performed using animal (rabbit) blood treated with trisodium citrate to prevent coagulation. CaCl2 solutions of varying concentrations are added to citrated blood, producing Ca(2+) ions to re-establish the coagulation cascade and mimic different blood coagulation abilities in vitro. Blood samples are dispensed into a paper-based LFA device consisting of sample pad, analytical membrane and wicking pad. The porous nature of the cellulose membrane separates the aqueous plasma component from the large blood cells. Since the viscosity of blood changes with its coagulation ability, the distance RBCs travel in the membrane in a given time can be related to the blood clotting time. The distance of the RBC front is found to decrease linearly with increasing CaCl2 concentration, with a travel rate decreasing from 3.25 mm min(-1) for no added CaCl2 to 2.2 mm min(-1) for 500 mM solution. Compared to conventional plasma clotting analyzers, the LFA device is much simpler and it provides a significantly larger linear range of measurement. Using the red colour of RBCs as a visible marker, this approach can be utilized to produce a simple and clear indicator of whether the blood condition is within the appropriate range for the patient's condition.

Triggered release of molecules across droplet interface bilayer lipid membranes using photopolymerizable lipids.

A combination of nonpolymerizable phospholipids (DPPC or DPhPC) and a smaller amount of cross-linking photopolymerizable phospholipids (23:2 DiynePC) is incorporated in an unsupported artificial lipid bilayer formed using the droplet interface bilayer (DIB) approach. The DIB is formed by contacting lipid monolayer-coated aqueous droplets against each other in a dodecane-lipid medium. Cross-linking of the photopolymerizable lipids incorporated in the DIB was obtained by exposure to UV-C radiation (254 nm), resulting in pore formation. The effect of cross-linking on the DIB properties was characterized optically by measuring the diffusion of selectively encapsulated dye molecules (calcein) from one droplet of the DIB to the other droplet. Changes in DIB conductivity due to UV-C exposure were investigated using current-voltage (I-V) measurements. The leakage of dye molecules across the DIB and the increase in DIB conductivity after UV-C exposure indicates the formation of membrane pores. The results indicate that the DIB approach offers a simple and flexible platform for studying phototriggered drug delivery systems in vitro.

Deactivating chemical agents using enzyme-coated nanofibers formed by electrospinning.

The coaxial electrospinning technique was investigated as a novel method to create stabilized, enzyme-containing fibers that have the potential to provide enhanced protection from chemical agents. Electrospinning is a versatile technique for the fabrication of polymer fibers with large length (cm to km): diameter (nm to µm) aspect ratios. The large surface to volume ratios, along with the biofriendly nature of this technique, enables the fabrication of fiber mats with high enzyme concentrations, which amplify the catalytic activity per unit volume of membrane. Blended composite (single-source) fibers incorporate enzyme throughout the fiber, which may limit substrate accessibility to the enzyme. In contrast, core/sheath fibers can be produced by coaxial electrospinning with very high enzyme loading (>80%) in the sheath without noticeable loss of enzymatic activity. Several core-sheath combinations have been explored with the toxin-mitigating enzyme DFPase in order to achieve fibers with optimum properties. The concentration of fluoride released, normalized for the amount of protein incorporated into the sheath, was used as a measure of the enzyme activity versus time. The coaxial core/sheath combination of PEO and DFPase produced the highest activity (~7.3 mM/mg).

Complementary electrowetting devices on plasma-treated fluoropolymer surfaces.

A reversal of the normal two-fluid competitive (water vs oil) electrowetting (nEW) on dielectric has been achieved by plasma irradiation of the normally hydrophobic fluoropolymer followed by thermal annealing. This process first renders the surface hydrophilic and then returns it to its normal hydrophobic properties as measured by water droplet contact and rolling angles. In the plasma-irradiated and annealed EW device (pEW), the normal two-fluid EW action is reversed after an initial charging step, with the oil layer being displaced at zero voltage and being returned at high voltage. A possible explanation of this effect is a plasma-induced modification of the fluoropolymer, rendering it more susceptible to charge injection and trapping at high voltage. nEW and pEW devices exhibit complementary EW operation, as verified by oil movement and optical transmission. This method can lead to low-power operation of two-fluid EW devices.

Role of surfactants in the interaction of dye molecules in natural DNA polymers.

Solutions and powders formed from salmon sperm deoxyribonucleic acid (DNA) reacted with the cationic surfactant cetyltrimethylammonium chloride (CTMA-Cl) incorporated fluorescent rhodamine molecules: anionic sulforhodamine 640 (SRh) or cationic/zwitterionic rhodamine 640 perchlorate (RhP). The role of the cationic surfactant in the interaction between rhodamine dye and DNA-surfactant molecules has been investigated in both solution and solid state using optical spectroscopy and electrophoresis. Unexpectedly, the dye molecules did not interact directly with DNA, rather the DNA double helix acted as a template for the interaction between dye molecules and CTMA in the DNA/CTMA complex. The SRh and RhP molecules yield different fluorescence characteristics with increasing DNA/CTMA amount, indicating different configurations between the CTMA ligands.

Photoluminescence and lasing from deoxyribonucleic acid (DNA) thin films doped with sulforhodamine.

Thin solid films of salmon deoxyribonucleic acid (DNA) have been fabricated by treatment with a surfactant and used as host for the laser dye sulforhodamine (SRh). The DNA films have an absorption peak at approximately 260 nm owing to absorption by the nitrogenous aromatic bases. The SRh molecules in the DNA films have absorption and emission peaks at 578 and 602 nm, respectively. The maximum emission was obtained at approximately 1 wt. % SRh in DNA, equivalent to approximately 100 DNA base pairs per SRh molecule. A distributed feedback grating structure was fabricated on a SiO(2)-Si substrate using interference lithography. The grating period of 437 nm was selected, corresponding to second-order emission at the amplified spontaneous emission wavelength of 650 nm. Lasing was obtained by pumping with a doubled Nd:YAG laser at 532 nm. The lasing threshold was 3 microJ, corresponding to approximately 30 microJ/cm(2) or 4 kW/cm(2). The emission linewidth decreased from approximately 30 nm in the amplified spontaneous emission mode to <0.4 nm (instrument limited) in the lasing mode. The slope efficiency of the lasing was approximately 1.2%.

Growth temperature dependence of optical modal gain and loss in GaN:Eu active medium on Si.

The dependence of optical modal gain and loss on GaN:Eu growth temperature is reported. GaN:Eu thin films were grown on Si substrates with AlGaN transition and cladding layers at temperatures ranging from 600 degrees C to 850 degrees C. The modal gain and loss in the GaN:Eu layer were a strong function of the optically active Eu atomic concentration and of the interface quality between the active layer and the top cladding layer, which in turn depended on the growth temperature. Optimum optical properties of maximum modal gain of ~ 100 cm(-1) and minimum loss of ~ 46 cm(-1) were obtained for growth at 800 degrees C.

High-Density Er-Implanted GaN Optical Memory Devices.

Upconversion emission has been obtained from Er-focused ion-beam (FIB) implanted GaN. Visible green emission at the 522- and 546-nm range were excited with infrared (IR) laser sources at either 840 or 1000 nm, or with both lasers simultaneously. By implanting closely spaced patterns with the FIB, we demonstrated the concept of storing data in Er-implanted GaN. Information stored as data bits consists of patterns of implanted locations as logic 1 and unimplanted locations as logic 0. The photon upconversion process in Er ions is utilized to read the stored information. This process makes use of the IR lasers to excite visible emission. The integrated upconversion emission power was measured to be ~40 pW when pumped by a 840-nm laser at 265 mW and by a 1000-nm laser at 208 mW. Patterns as small as 0.5 mum were implanted and read. Three-dimensional optical memory based on rare-earth-doped semiconductors could in theory approach a storage capacity of 10(12) bits/cm(3).