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 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…
Introduction This experiment is a continuation of previous work, cited below. Instead of urea spiked 1x PBS, whole heparinized bovine blood was pushed through the device over a period of four hours to then perform a urea assay on the separated and collected plasma. Please refer to the post below regarding the experimental setup that…
Introduction These results are thanks to assistance from Sam Walker and his previous posts regarding urea clearance that helped lay the foundation of this experiment. “Single Slot Inverted-Chip and Double Slot: Single-Pass Dialysis Devices” (Walker 2018) https://trace-bmps.org/single-slot-inverted-chip-and-double-slot-single-pass-dialysis-devices/ More information on the Liftoff membranes can be found in the article cited below: “Lift-off of large-scale…
After continually seeing variability in assays using the microfluidic silicon membrane canalicular array (µSiM-CA) and developing a new technique to visualize bacterial division in the µSiM-CA, discussed here, we wanted to improve and stabilize the performance of our assays. Currently, the µSiM-CA is used by our lab, the Gill lab and the Dunman lab to…
Recently, several studies on Staphylococcus aureus (S. aureus) have been hindered due to poor performance from assays using the µSiM-Canalicular Array (CA). As discussed here, we attempted to improve assay performance by coating our membranes with fibronectin. This did not work well and instead left us in a state of confusion as we did not…
This post describes my second attempt at developing a protocol for viral genome extraction in large volumes of waste water. Like our first attempt, described here, we looked to improve upon the protocol Direct wastewater RNA extraction via the “Milk of Silica (MoS)” method – A companion method to “Sewage, Salt, Silica and SARS-CoV-2 (4S)”….
Parallel with my efforts in the Osteomyelitis project to coat our membranes with polyurethane acrylate, I began working with the Gill lab on improving the performance of our uSiM platform as a propagation assay for Staphylococcus aureus. In the past we have gotten a lot of good data using this assay, but its performance has…
Since I discovered a dilution of polyurethane acrylate (PUA) which coats well (described here), I have restarted my efforts towards etching the PUA. Described below are my most recent attempts towards creating a PUA coating with a smooth surface with a 1 um in thickness. Unfortunately, it has proven harder to maintain a smooth surface…
Introduction The blood-brain barrier (BBB) plays an important role in maintaining brain chemistry by restricting the permeability of small molecules and cells. Inflammatory diseases may disrupt the BBB and promote the development of neurological disorders [1]. One potential mechanism of BBB disruption is the degradation of the extracellular matrix proteins that make up the vascular…