R61: Slide Decks for 6 Month Progress Report
PDF with all slides: R61 HL154249-01(6 Month Progress)
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Monolayer Culture of mouse CMECs on μSIM A-Line Devices
Introduction In the Glading Lab we study a rare disease called Cerebral Cavernous Malformations (CCMs). In this disease, brain capillaries develop greatly enlarged lumens and a disrupted blood-brain barrier. A genetic cause of the disease is the presence of a mutation in KRIT1 that causes loss of KRIT1 protein function in the cells. KRIT1 binds…
Bacterial Inflammatory Stimulation of the μSiM-BBB Model
Introduction Sepsis refers to an uncontrolled systemic inflammatory response which can result from bacterial blood infections. We have long been interested in developing a sepsis model using the μSiM-BBB platform because the neuroinflammation stemming from sepsis can cause severe BBB disruption. For the majority of our stimulation experiments we have relied on cytomix, a cocktail…
COMSOL simulation on sampling method to measure membrane permeability
Background We at Michigan have been aiming to perform sequential small volume sampling coupled with our ultrasensitive digital immunoassay to provide longitudinal data from the uSIM. We have encountered repeatability problems as well as low molecule abundancy with shortly incubated samples collected from the bottom microfluidic channel. Therefore, COMSOL simulation work has been done to…
QC on Dual Scale Membrane (lot 1383)_from Bern
We recently received 100 Dual Scale Membrane (lot 1383). As noted in Dan’s post we also had same issues regarding mix up in the labeling. In the gel box, 2 boxes (25 membranes/box) are labeled 0.625% porosity and 2boxes are labeled 1.25% porosity, thus we supposed to have 50 membranes for both porosity. However 1…
Collagen IV is deposited below human umbilical vein endothelial cells
Introduction My previous post focused on attempts to reproducibly generate collagen IV with human umbilical vein endothelial cells (HUVECs) so that we would be able to establish protocols for basement membrane protein detection and degradation measurement. Following my presentation on November 18 2020, there were some questions regarding whether or not the observed collagen IV…
Multiplexed sequential brain endothelial cell cytokine secretion measurement from μSIM
Background Previously, we have demonstrated multiplexed digital sensing on MCP1, KC and TNFα secreted by RAW 264.7 cells stimulated in conventional cell culturing well plates. We observed decent TNFα and strong MCP1 response but no KC secretion from RAW cells upon 100ng/ml of LPS stimulation overnight. The work verified our platform’s capability of multiplex cytokine…