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…
Category: NRG
To Bead or not to Bead: Struggling to Detect tRNA Binding to Silica Particles towards Wastewater Genome Extraction
Posted in NRG
Physiological Facade: Fibronectin coating membranes in an attempt to improve the performance of Staphylococcus aureus propagation assays using the µSiM
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…
Posted in NRG
Flabbergrassted: Persistent Surface Roughness in Polyurethane Acrylate Etches
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…
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Fibronectin and Friends: Confirming the Presence of Multiple Vascular Basement Membrane Components in the μSiM-hNVU
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…
Does the Shoe Fit? Re-evaluating and Re-discovering an Amenable PUA Solution
Since we last discovered our polyurethane acrylate (PUA) has a shelf life (described here) and received a new solution of PUA, we have been trying to replicate the results that we had before. Unfortunately, we have realized that this is…
Posted in NRG
Optimizing IMR90-4 Coculture BBB Model
Introduction While we have optimized coculture models for commercial brain endothelial cells and pericytes, we needed to do the same for iPSC-derived cocultures. In this post, I show the optimization process using IMR90-4-derived EECM-BMEC-like cells [1] and brain pericyte-like cells (BPLC)…
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Optimizing Cytomix Stimulation on Mono- and Coculture BBB Models
Introduction The µSiM BBB model is being developed at UR with the goal to understand the impact of sepsis on the brain. Here, we use an EECM-BMEC-like cell (iSPC-derived brain endothelial cells) [1] monoculture model and an EECM-BMEC-like cell and…
Merging Man with Machine: A Computer Vision Aided Leukocyte Transmigration Assay
Introduction High throughput imaging techniques in conjunction with in vitro micro-physiological systems (MPS) allow for comprehensive exploration of physiologically relevant phenomena in a living 3D mimetic of a tissue microenvironment. Prior work in this lab has readily demonstrated that vascular…
Updates on Modeling the Vascular Basement Membrane in the μSiM-hNVU
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…
Fabrication the Nano-pocket membrane (update)
The fabrication of nano pocket membrane to capture and release the EV faced challenges during the etching step. There was a suggestion after the last NRG post to use RIE-2000 located in the UR. This post shows the update on…
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