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…
Category: 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)…
Posted in NRG
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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Crazy Colorful Counting Conundrum: From Counts to Colocalization
Summary We look at nanoparticle counting and colocalization after an ALine device membrane capture and compare IMARIS and ImageJ methods with an example. Procedures for particle counting and colocalization for IMARIS are discussed and a walkthrough is provided An example…
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The Enemy Within: Optimizing an OLCN PUA Coated Microporous Membrane Mimetic and Exposing an Unlikely Saboteur
The battle for soft membrane coatings wages on as we continue to optimize polyurethane acrylate (PUA) coatings for our osteomyelitis project, whose background is described here and here and initial progress is described here. Since our last update, we have…
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Aliens, Voodoo, or Magic? How does DNA Extraction via Silica Adsorption Actually Work?
The first DNA extraction was done over 150 years ago in 18691. Since then, the process has been refined and expanded to include multiple physical and or chemical methods. For the waste water epidemiology project (described here), we will be…
Posted in NRG
Daily In-Situ Permeability using mouse CMECs
Introduction In the Glading Lab we study Cerebral Cavernous Malformations (CCMs). In CCMs, brain capillaries develop enlarged lumens and a disrupted blood-brain barrier. The presence of a mutation in KRIT1 causes loss of KRIT1 protein function in the endothelial cells.…
