Accelerated colorimetric immunosensing using surface-modified porous monoliths and gold nanoparticles.

A rapid and sensitive immunoassay platform integrating polymerized monoliths and gold nanoparticles (AuNPs) has been developed. The porous monoliths are photopolymerized in situ within a silica capillary and serve as solid support for high-mass transport and high-density capture antibody immobilization to create a shorter diffusion length for antibody-antigen interactions, resulting in a rapid assay and low reagent consumption. AuNPs are modified with detection antibodies and are utilized as signals for colorimetric immunoassays without the need for enzyme, substrate and sophisticated equipment for quantitative measurements. This platform has been verified by performing a human IgG sandwich immunoassay with a detection limit of 0.1 ng ml-1. In addition, a single assay can be completed in 1 h, which is more efficient than traditional immunoassays that require several hours to complete.

Rapid and inexpensive blood typing on thermoplastic chips.

A portable and cost-effective colorimetric diagnostic device was fabricated for rapid ABO and Rh blood typing. Using microfluidic construction on a thermoplastic chip, blood antibodies were preloaded into a reaction channel and exposed to blood samples to initiate a haemagglutination reaction. Downstream high-aspect ratio filters, composed of 2 µm high microslits, block agglutinated red blood cells (RBCs) to turn the reaction channel red, indicating the presence of the corresponding blood antigen. Users manually actuate the blood sample using a simple screw pump that drives the solution through serpentine reaction channels and chaotic micromixers for maximum interaction of the preloaded antibodies with the blood sample antigens. Mismatched RBCs and antibodies elute from the channel into an outlet reservoir based on the rheological properties of RBCs with no colorimetric change. As a result, unambiguous blood typing tests can be distinguished by the naked eye in as little as 1 min. Blood disorders, such as thalassemia, can also be distinguished using the device. The required blood volume for the test is just 1 µL, which can be obtained by the less invasive finger pricking method. The low reagent consumption, manual driving force, low-cost of parts, high yield, and robust fabrication process make this device sensitive, accurate, and simple enough to use without specialized training in resource constrained settings.

Release of FITC-BSA from poly(l-lactic acid) microspheres analysis using flow cytometry.

In this investigation, biodegradable polymer poly(L-lactic acid) (PLA) microspheres were prepared by the W(1)/O/W(2) solvent evaporation method. The inner phase was aqueous solution (W(1)) that contained bovine serum albumin that was labeled with fluorescein isothiocyanate (FITC-BSA). PLA was dissolved in chloroform with emulsifier sorbitan monooleate (span 80) as the dispersed phase (O). These two solutions (W(1)/O) were emulsified by a homogenizer to form a primary emulsion. Polyvinyl alcohol (PVA) used as surfactant, was applied in the formation of microspheres (W(2)). 0.5% (w/v) PLA was stirred at 3000 rpm using a homogenizer. Microspheres with sizes of up to around 10 µm were produced. These microspheres were separated by the glycerol gradient method, and take microspheres at part of 25% glycerol gradient concentration was analyzed by flow cytometry, indicating a more homogeneous particle size distribution than that not separated. The microspheres were degraded using several enzymes, and around 40% was degraded by 72 h. This result reveals the effectiveness of drug delivery by PLA microspheres, which was evaluated by performing a drug release test and flow cytometric analysis. The FITC-BSA concentration in the supernatant increased with the experimental time. At the phagocytosis experiments, encapsulated with FITC-BSA drug of microspheres can be used by the cell, as particle size approximately 1 µm.