PRADA: Portable Reusable Accurate Diagnostics with nanostar Antennas for multiplexed biomarker screening.

Precise monitoring of specific biomarkers in biological fluids with accurate biodiagnostic sensors is critical for early diagnosis of diseases and subsequent treatment planning. In this work, we demonstrated an innovative biodiagnostic sensor, portable reusable accurate diagnostics with nanostar antennas (PRADA), for multiplexed biomarker detection in small volumes (~50 µl) enabled in a microfluidic platform. Here, PRADA simultaneously detected two biomarkers of myocardial infarction, cardiac troponin I (cTnI), which is well accepted for cardiac disorders, and neuropeptide Y (NPY), which controls cardiac sympathetic drive. In PRADA immunoassay, magnetic beads captured the biomarkers in human serum samples, and gold nanostars (GNSs) "antennas" labeled with peptide biorecognition elements and Raman tags detected the biomarkers via surface-enhanced Raman spectroscopy (SERS). The peptide-conjugated GNS-SERS barcodes were leveraged to achieve high sensitivity, with a limit of detection (LOD) of 0.0055 ng/ml of cTnI, and a LOD of 0.12 ng/ml of NPY comparable with commercially available test kits. The innovation of PRADA was also in the regeneration and reuse of the same sensor chip for ~14 cycles. We validated PRADA by testing cTnI in 11 de-identified cardiac patient samples of various demographics within a 95% confidence interval and high precision profile. We envision low-cost PRADA will have tremendous translational impact and be amenable to resource-limited settings for accurate treatment planning in patients.

Profiling and Imaging Ion Mobility-Mass Spectrometry Analysis of Cholesterol and 7-Dehydrocholesterol in Cells Via Sputtered Silver MALDI.

Profiling and imaging of cholesterol and its precursors by mass spectrometry (MS) are important in a number of cholesterol biosynthesis disorders, such as in Smith-Lemli-Opitz syndrome (SLOS), where 7-dehydrocholesterol (7-DHC) is accumulated in affected individuals. SLOS is caused by defects in the enzyme that reduces 7-DHC to cholesterol. However, analysis of sterols is challenging because these hydrophobic olefins are difficult to ionize for MS detection. We report here sputtered silver matrix-assisted laser desorption/ionization (MALDI)-ion mobility-MS (IM-MS) analysis of cholesterol and 7-DHC. In comparison with liquid-based AgNO3 and colloidal Ag nanoparticle (AgNP), sputtered silver NP (10-25 nm) provided the lowest limits-of-detection based on the silver coordinated [cholesterol + Ag](+) and [7-DHC + Ag](+) signals while minimizing dehydrogenation products ([M + Ag-2H](+)). When analyzing human fibroblasts that were directly grown on poly-L-lysine-coated ITO glass plates with this technique, in situ, the 7-DHC/cholesterol ratios for both control and SLOS human fibroblasts are readily obtained. The m/z of 491 (specific for [7-DHC + (107)Ag](+)) and 495 (specific for [cholesterol + (109)Ag](+)) were subsequently imaged using MALDI-IM-MS. MS images were co-registered with optical images of the cells for metabolic ratio determination. From these comparisons, ratios of 7-DHC/cholesterol for SLOS human fibroblasts are distinctly higher than in control human fibroblasts. Thus, this strategy demonstrates the utility for diagnosing/assaying the severity of cholesterol biosynthesis disorders in vitro.

Growth of Solid and Hollow Gold Particles through the Thermal Annealing of Nanoscale Patterned Thin Films.

Through thermally annealing well-arrayed, circular, nanoscale thin films of gold, deposited onto [111] silicon/silicon dioxide substrates, both solid and hollow gold particles of different morphologies with controllable sizes were obtained. The circular thin films formed individual particles or clusters of particles by tuning their diameter. Hollow gold particles were characterized by their diameter, typically larger than 400 nm; these dimensions and properties were confirmed by cross-section scanning electron microscopy. Hollow gold particles also exhibited plasmonic field enhancement under photoemission electron microscopy. Potential growth mechanisms for these structures were explored.

Experimental characterization of a metal-oxide-semiconductor field-effect transistor-based Coulter counter.

We report the detailed characterization of an ultrasensitive microfluidic device used to detect the translocation of small particles through a sensing microchannel. The device connects a fluidic circuit to the gate of a metal-oxide-semiconductor field-effect transistor (MOSFET) and detects particles by monitoring the MOSFET drain current modulation instead of the modulation in the ionic current through the sensing channel. The minimum volume ratio of the particle to the sensing channel detected is 0.006%, which is about ten times smaller than the lowest detected volume ratio previously reported in the literature. This volume ratio is detected at a noise level of about 0.6% of the baseline MOSFET drain current, clearly showing the amplification effects from the fluidic circuits and the MOSFETs. We characterize the device sensitivity as a function of the MOSFET gate potential and show that its sensitivity is higher when the MOSFET is operating below its threshold gate voltage than when it is operating above the threshold voltage. In addition, we demonstrate that the device sensitivity linearly increases with the applied electrical bias across the fluidic circuit. Finally, we show that polystyrene beads and glass beads with similar sizes can be distinguished from each other based on their different translocation times, and the size distribution of microbeads can be obtained with accuracy comparable to that of direct scanning electron microscopy measurements.