There is a strong interest in the use of biopolymers in the electronic and biomedical industries, mainly towards low-cost applications. The possibility of developing entirely new kinds of products based on cellulose is of current interest, in order to enhance and to add new functionalities to conventional paper-based products. We present our results towards the development of paper-based microfluidics for molecular diagnostic testing. Paper properties were evaluated and compared to nitrocellulose, the most commonly used material in lateral flow and other rapid tests. Focusing on the use of paper as a substrate for microfluidic applications, through an eco-friendly wax-printing technology, we present three main and distinct colorimetric approaches: (i) enzymatic reactions (glucose detection); (ii) immunoassays (antibodies anti-Leishmania detection); (iii) nucleic acid sequence identification (Mycobacterium tuberculosis complex detection). Colorimetric glucose quantification was achieved through enzymatic reactions performed within specific zones of the paper-based device. The colouration achieved increased with growing glucose concentration and was highly homogeneous, covering all the surface of the paper reaction zones in a 3D sensor format. These devices showed a major advantage when compared to the 2D lateral flow glucose sensors, where some carryover of the coloured products usually occurs. The detection of anti-Leishmania antibodies in canine sera was conceptually achieved using a paper-based 96-well enzyme-linked immunosorbent assay format. However, optimization is still needed for this test, regarding the efficiency of the immobilization of antigens on the cellulose fibres. The detection of Mycobacterium tuberculosis nucleic acids integrated with a non-cross-linking gold nanoprobe detection scheme was also achieved in a wax-printed 384-well paper-based microplate, by the hybridization with a species-specific probe. The obtained results with the above-mentioned proof-of-concept sensors are thus promising towards the future development of simple and cost-effective paper-based diagnostic devices.
In this paper we employed phospholipid Langmuir monolayers as membrane models to probe interactions with chitosan. Using a combination of surface pressure--area and surface potential--area isotherms and rheological measurements with the pendent drop technique, we observed that chitosan interacts with phospholipid molecules at the air-water interface. We propose a model in which chitosan interacts with the phospholipids mainly through electrostatic interactions, but also including H-bonding and hydrophobic forces, depending on the phospholipid packing density. At large areas per molecule, chitosan in the subphase adsorbs onto the monolayer, expanding it. At small areas per molecule, chitosan is located in the subsurface. Indeed, a mixed chitosan-phospholipid monolayer can be transferred onto solid supports, even at high surface pressures. The effects of chitosan on the viscoelastic properties of phospholipid monolayers may be taken as evidence for the ability of chitosan to disrupt cell membranes.
The results of the search for the optimal experimental conditions for ultrasentitive chemical analysis of 1-naphthalenethiol (1-NAT) and 2-naphthalenethiol (2-NAT) using surface-enhanced Raman scattering (SERS) are discussed. The report begins with a review of the vibrational spectra, including infrared and Raman spectra of the target molecules, and the interpretation of the observed frequencies aided by local density functional theory (DFT) calculations at the B3LYP/6-311G(d,p) level of theory. Several metal nanostructures were tested for SERS activity, including island films and colloids of silver, gold and copper. Correspondingly, the most effective laser line for excitation in the visible and near infrared region was sought. The achieved detection limit for 1-naphthalenethiol, and for 2-naphthalenethiol, on silver nanostructures is in the zeptomole regime.
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