Fabrication and Comparative Quantitative Analysis of Plasmonic-Polymer Nanocomposites as Optical Platforms.

Plasmonic-polymer nanocomposites can serve as a multifunctional platform for a wide range of applications such as biochemical sensing and photothermal treatments, where they synergistically benefit from the extraordinary optical properties of plasmonic nanoparticles (NPs) and biocompatible characteristics of biopolymers. The field translation of plasmonic-polymer nanocomposites requires design rules for scalable and reproducible fabrication with tunable and predictable optical properties and achieving the best performance. The optical properties of NPs and the optimal analytical performance of nanocomposites could be affected by many fabrication parameters, but a fundamental understanding of such parameters is still minimal. Herein, we systematically investigated the NP distribution and their optical properties in gold nanostar (GNS)-polymer nanocomposites as a function of GNS concentration, polymer identity, and the method of GNS incorporation into a polymer matrix. We performed a comprehensive analysis of the single-particle scattering spectra of GNS incorporated into agarose gel and chitosan hydrogels via embedding and surface deposition, using dark-field spectroscopy. While relative GNS concentration affects the GNS scattering property distribution in both polymer matrices, chemical interactions between a polymer matrix and GNS is the key determinant of the GNS stability and homogenous distribution in nanocomposites. When GNS are embedded in a polymer matrix and there are stronger chemical interactions between GNS and a polymer, significantly less aggregation and a more homogenous distribution of GNS, which leads to a larger percentage of GNS optical property preservation, were observed at all the concentrations. In a proof-of-concept surface-enhanced Raman spectroscopy (SERS) study, we observed that the SERS detection efficiency is dictated by the analyte accessibility of GNS, which is governed by the polymer matrix porosity, polymer-GNS interactions, and other polymer physical characteristics. This work presents the interplay between key fabrication parameters and foundational design parameters for more predictable and reliable fabrication of plasmonic-polymer nanocomposites as an optical platform.

Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles.

Direct detection of genetic biomarkers in body fluid lysate without target amplification will revolutionize nucleic acid-based diagnostics. However, the low concentration of target sequences makes this goal challenging. We report a method for direct detection of pathogen RNA in blood lysate using a bioassay using surface-enhanced Raman spectroscopy (SERS)-based detection integrated in a "lab-in-a-stick" portable device. Two levels of signal enhancement were employed to achieve the sensitivity required for direct detection. Each target sequence was tagged with an ultrabright SERS-encoded nanorattle with ultrahigh SERS signals, and these tagged target sequences were concentrated into a focused spot for detection using hybridization sandwiches with magnetic microbeads. Furthermore, the washing process was automated by integration into a "lab-in-a-stick" portable device. We could directly detect synthetic target with a limit of detection of 200 fM. More importantly, we detected plasmodium falciparum malaria parasite RNA directly in infected red blood cells lysate. To our knowledge, this is the first report of SERS-based direct detection of pathogen nucleic acid in blood lysate without nucleic acid extraction or target amplification. The results show the potential of our integrated bioassay for field use and point-of-care diagnostics.

SERS-based approaches toward genetic profiling.

In the last decade surface-enhanced Raman scattering (SERS) has experienced an important resurgence, and as a consequence it has seen wide application in the biological field, especially for DNA identification. SERS-based DNA detection can be carried out directly and indirectly and, in the latter approach, it relies on the use of SERS tags, whose role is to indirectly prove the recognition and binding of a specific oligonucleotide sequence. Herein, the role of SERS tags is analyzed focusing specifically on the use of DNA identification for genetic profiling.