When we first began developing a standardized protocol for μSiM-CA bacterial assays (described here), bad assay performance led some to believe our issues were due to availability of pores within the membranes inside of the μSiMs. At the same time, I developed a visualization protocol to view bacterial transmission across the membrane (described here). While…
Introduction When searching for a flow control system for microfluidic cell culture, we look to balance the ease of use, accessibility, and precision of fluidic control. For instance, researchers have explored different powered actuation methods for miniaturized precise proportional fluid control. These actuation methods include electromechanical, electrostatic, electromagnetic, piezoelectric, and thermal-based. The necessary power sources,…
Introduction This post is meant to serve as an update for some of the more recent work completed on the CAD-LB (catch and display for liquid biopsy) platform. The CAD-LB is intended to be used as a digital biomarker assay for interrogating small extracellular vesicles (sEVs). Since transitioning our assay to the µSIM device we…
Introduction The membrane on uSiM platform is inert, therefore, coating with extracellular matrix (ECM) proteins is essential to promote attachment and growth of adhesion cell lines on the uSiM. Besides promoting proper attachment on the membrane, the ECM proteins dramatically impact cell growth, differentiation, and functions. Therefore, careful consideration must be given in selecting the…
Attached is a PDF presented to the NIH at our annual update meeting that included other funded project teams. R61_R33HL154249 11-22 Update (Low Res)
While imaging GFP tagged Staphylococcus aureus on our membranes within µSiM-CA devices, the question of whether or not we can differentiate between bacteria on the top and bottom of the membrane versus within the membrane has persisted. We know that during our experiments bacteria exist in all three of these places, therefore the question really is whether…
Introduction Tissue barriers separate different compartments of body organs. These barriers regulate the absorption of nutrients, maintain homeostasis, and prevent toxins from entering. In vitro models of tissue barriers have been developed to understand human diseases and investigate drug toxicity and effectiveness. In order to investigate cell-cell junctions and barrier maturity created on these models,…
Since our attempt to standardize the Staphylococcus aureus µSiM-CA division assay, described here, we have moved towards investigating biological reasons as to why our assay performance is not where we want it to be. While biofilms have been suspect from the beginning, we have also thought that bacterial transmission requires two main steps: 1) Pore discovery…
Introduction The capability of the m-μSiM for establishing an hiPSC-derived blood-brain barrier (BBB) in bioengineering and bioscience laboratories has been shown previously [1]. To further demonstrate interlaboratory agreement and successful dissemination of components and protocols, m-μSiM was used at RIT (Abhyankar lab) to establish an hiPSC-BMECs monolayer. Method All reagents and steps were used based…
Introduction Studies have shown that the shear-stimulation of endothelial cells results in the upregulation of the shear-sensitive transcription factor Kruppel-like factor 2 (KLF2). KLF2 plays a critical role in the regulation of endothelial activity such as the regulation of vascular tone, anti-inflammatory responses, and antithrombotic functions. KLF2 is also responsible for the upregulation of endothelial…