A review of DNA and RNA sample preparation techniques from cell lysis to nucleic acid purification and how they advance on-chip microfluidic nucleic acid analys

A chemically patterned microfluidic paper-based analytical device (C-µPAD) is developed to create fluidic networks by forming hydrophobic barriers using chemical vapor deposition (CVD) of trichlorosilane (TCS) on a chromatography paper. By controlling temperature, pattern size, and CVD duration, optimal conditions were determined by characterizing hydrophobicity, spreading patterns, and flow behavior on various sized fluidic patterns. With these optimal conditions, we demonstrated glucose assay, immunoassay, and heavy metal detection on well-spot C-µPAD and lateral flow C-µPAD. For these assays, standard curves showing correlation between target concentration and gray intensity were obtained to determine a limit of detection (LOD) of each assay. For the glucose assays on both well-spot C-µPAD and lateral flow C-µPAD, we achieved LOD of 13 mg/dL, which is equivalent to that of a commercial glucose sensor. Similar results were obtained from tumor necrosis factor alpha (TNFα) detection with 3 ng/mL of LOD. For Ni detection, a colorimetric agent was immobilized to obtain a stationary and uniform reaction by using thermal condensation coupling method. During the immobilization, we successfully functionalized amine for coupling the colorimetric agent on the C-µPAD and detected as low as 150 μg/L of Ni. These C-µPADs enable simple, rapid, and cost-effective bioassays and environmental monitoring, which provide practically relevant LODs with high expandability and adaptability.

Animals are commonly used for pharmacokinetic studies which are the most frequent events tested during ocular drug development and preclinical evaluation. Inaccuracy, cost, and ethical criticism in these tests have created a need to construct an in vitro model for studying corneal constraints. In this work, a porous membrane embedded microfluidic platform is fabricated that separates a chip into an apical and basal side. After functionalizing the membrane surface with fibronectin, the membrane's mechanical and surface properties are measured to ensure correct modeling of in vivo characteristics. Immortalized human corneal epithelial cells are cultured on the membrane to create a microengineered corneal epithelium-on-a-chip (cornea chip) that is validated with experiments designed to test the barrier properties of the human corneal epithelium construct using model drugs. A pulsatile flow model is used that closely mimics the ocular precorneal constraints and is reasonable for permeability analysis that models in vivo conditions. This model can be used for preclinical evaluations of potential therapeutic drugs and to mimic the environment of the human cornea.