9 research outputs found
Surface enhanced Raman scattering (SERS) based microfluidics for single cell analysis
The integration of surface enhanced Raman scattering (SERS) with droplet microfluidics has the potential to improve our understanding of cellular systems. Herein, we present the first application of SERS droplet microfluidics for single cell analysis. A microfluidic device was used to encapsulate single prostate cancer cells and wheat germ agglutin (WGA) functionalized SERS nanoprobes in water-in-oil droplets that were subsequently locked into a storage droplet array for spectroscopic investigation. The stationary droplets enabled the rapid identification of SERS regions of interest in live cancer cells by allowing collection of âfastâ coarse maps over an area of several mm2 followed by âslowerâ detailed interrogation of the identified hotspots. We demonstrate SERS at cellular resolution via a proof-of-concept assay that detects glycan expression on the surface of prostate cancer cells using WGA modified metallic nanoparticles. The data illustrates the potential of SERS optofluidic systems for high-throughput cell screening and illustrates a previously unobserved high degree of cell-to-cell variability in the size and number of glycan islands
Aerosol Microdroplets Exhibit a Stable pH Gradient
Suspended aqueous aerosol droplets (\u3c50 ÎŒm) are microreactors for many important atmospheric reactions. In droplets and other aquatic environments, pH is arguably the key parameter dictating chemical and biological processes. The nature of the droplet air/ water interface has the potential to significantly alter droplet pH relative to bulk water. Historically, it has been challenging to measure the pH of individual droplets because of their inaccessibility to conventional pH probes. In this study, we scanned droplets containing 4-mercaptobenzoic acidâfunctionalized gold nanoparticle pH nanoprobes by 2D and 3D laser confocal Raman microscopy. Using surface-enhanced Raman scattering, we acquired the pH distribution inside approximately 20-ÎŒm-diameter phosphate-buffered aerosol droplets and found that the pH in the core of a droplet is higher than that of bulk solution by up to 3.6 pH units. This finding suggests the accumulation of protons at the air/water interface and is consistent with recent thermodynamic model results. The existence of this pH shift was corroborated by the observation that a catalytic reaction that occurs only under basic conditions (i.e., dimerization of 4-aminothiophenol to produce dimercaptoazobenzene) occurs within the high pH core of a droplet, but not in bulk solution. Our nanoparticle probe enables pH quantification through the cross-section of an aerosol droplet, revealing a spatial gradient that has implications for acid-baseâcatalyzed atmospheric chemistry
Nanomaterial enabled sensors for environmental contaminants
Abstract The need and desire to understand the environment, especially the quality of oneâs local water and air, has continued to expand with the emergence of the digital age. The bottleneck in understanding the environment has switched from being able to store all of the data collected to collecting enough data on a broad range of contaminants of environmental concern. Nanomaterial enabled sensors represent a suite of technologies developed over the last 15 years for the highly specific and sensitive detection of environmental contaminants. With the promise of facile, low cost, field-deployable technology, the ability to quantitatively understand nature in a systematic way will soon be a reality. In this review, we first introduce nanosensor design before exploring the application of nanosensors for the detection of three classes of environmental contaminants: pesticides, heavy metals, and pathogens
Position Effects Due to Chromosome Breakpoints that Map âŒ900 Kb Upstream and âŒ1.3 Mb Downstream of SOX9 in Two Patients with Campomelic Dysplasia
Campomelic dysplasia (CD) is a semilethal skeletal malformation syndrome with or without XY sex reversal. In addition to the multiple mutations found within the sex-determining region Yârelated high-mobility group box gene (SOX9) on 17q24.3, several chromosome anomalies (translocations, inversions, and deletions) with breakpoints scattered over 1 Mb upstream of SOX9 have been described. Here, we present a balanced translocation, t(4;17)(q28.3;q24.3), segregating in a family with a mild acampomelic CD with Robin sequence. Both chromosome breakpoints have been identified by fluorescence in situ hybridization and have been sequenced using a somatic cell hybrid. The 17q24.3 breakpoint maps âŒ900 kb upstream of SOX9, which is within the same bacterial artificial chromosome clone as the breakpoints of two other reported patients with mild CD. We also report a prenatal identification of acampomelic CD with male-to-female sex reversal in a fetus with a de novo balanced complex karyotype, 46,XY,t(4;7;8;17)(4qterâ4p15.1::17q25.1â17qter;7qterâ7p15.3::4p15.1â4pter;8pterâ8q12.1::7p15.3â7pter;17pterâ17q25.1::8q12.1â8qter). Surprisingly, the 17q breakpoint maps âŒ1.3 Mb downstream of SOX9, making this the longest-range position effect found in the field of human genetics and the first report of a patient with CD with the chromosome breakpoint mapping 3âČ of SOX9. By using the Regulatory Potential score in conjunction with analysis of the rearrangement breakpoints, we identified a candidate upstream cis-regulatory element, SOX9cre1. We provide evidence that this 1.1-kb evolutionarily conserved element and the downstream breakpoint region colocalize with SOX9 in the interphase nucleus, despite being located 1.1 Mb upstream and 1.3 Mb downstream of it, respectively. The potential molecular mechanism responsible for the position effect is discussed
Improved Quantitative SERS Enabled by Surface Plasmon Enhanced Elastic Light Scattering
The application of
surface-enhanced Raman spectroscopy (SERS) for
everyday quantitative analysis is hindered by the point-to-point variability
of SERS substrates that arises due to the heterogeneous distribution
of localized electromagnetic fields across a suite of plasmonic nanostructures.
Herein, we adopt surface-enhanced elastic scattering as a SERS internal
standard. Both elastic and inelastic (i.e., Raman) scattering are
simultaneously enhanced by a given âhot spotâ, and thus,
the surface-enhanced elastic scattering signal provides a localized
intrinsic internal standard that scales across all of the plasmon-enhanced
electromagnetic fields within a substrate. Elastically scattered light
originates from the amplified spontaneous emission (ASE) of the commercial
laser, leading to the formation of a low-wavenumber pseudo band that
arises from the interaction of the ASE and the edge filter. A theoretical
model was developed to illustrate the underlying mechanism supporting
this normalization approach. The normalized Raman signals are independent
of the incident laser intensity and the density of âhot spotsâ
for numerous SERS substrates. Following âhot-spotâ (HS)
normalization, the coefficient of variation for the tested SERS substrates
decreases from 10 to 60% to 2%â7%. This approach significantly
improves SERS quantitation of four chloroanilines and enables collection
of highly reproducible analyte adsorption results under both static
and dynamic imaging conditions. Overall, this approach provides a
simple means to improve SERS reproducibility without the need to use
additional chemicals as internal standards