Introduction Inflammatory diseases have the potential to disrupt blood vessel permeability. This has serious ramifications for the blood-brain barrier (BBB), which is the interface between the vasculature and the brain. A healthy BBB allows essential nutrients to pass through while excluding molecules that might alter brain chemistry. If inflammation were to increase the BBB’s permeability…
Introduction/Methods As part of our investigation into neutrophil transmigration on tissue chip systems featuring HUVEC monolayers, I have been working on developing code for both cell state analysis and tracking using Wolfram Mathematica (and occasionally Python). This project relies on semantic segmentation via machine learning and microscopy video corrections, both of which have been described…
Introduction Phase-contrast microscopy is widely utilised in live cell imaging, with pixel segmentation protocols in particular finding widespread usage as a means to computationally isolate cells from a tissue-culture background. Computer segmentation allows for the analysis of individual cell properties such as state or spatial location through automated or semi-automated techniques, ultimately attempting to avoid…
Introduction Selective leukocyte interactions with endothelial barriers are a characteristic component of the inflammatory response process. These spatio-temporal interactions can mechanically remodel local tissue, as well as damage healthy tissue with robust inflammation stemming from an over-reactive immune response. This is evident in many pathologies and can complicate severe conditions such as sepsis. Thus, it…
Introduction Read the manuscript here: Integrative Biology Publication Please refer to the linked manuscript for a detailed introduction. In short, we have been interested in characterizing the role of endothelial cell (EC) apicobasal polarity in inflammation. While usually reserved for the epithelium, ‘apicobasal polarity’ is a term describing a monolayer or cell with defined apical…
IntroductionThe ability to continuously sample and assay for the levels of soluble factors without disrupting the microenvironment is important to acquire sequential secretion profiles in BBB experiments on an uSIM module. Combining small volume sampling, buffer replenishment and sensitive pedELISA technology would enable longitudinal assays of proteins introduced to blood compartment or released from cells….
Vacuoles with pulling flow vs pushing flow This video shows an example of the first trials with the uSim flow cell (Tejas’ early versions) when creating laminar flow over a monolayer of BLECs – see Mossu et al, JCBFM 2018 – the issue we had to overcome was to avoid creating vacuoles in the endothelial…
Introduction A healthy blood-brain barrier is not permissive to most small molecules, but during inflammation, tight junctions weaken and the barrier becomes more permissive. We have developed an in situ permeability assay that tracks diffusion across a cell monolayer and membrane and into the bottom channel of the µSiM device. The assay tracks a fluorescent…
Link to BioRxiv paper The extracellular matrix in vivo is composed of fibers arranged with varying degrees of alignment in different directions. The extent of alignment (anisotropy) and the directionality of fibers are an indicator of breast cancer metastasis, influence T-cell motility, and guide endothelial cell alignment. Similar to the existence of chemical gradients and…
Just wanted to drop this data here in case it may be useful for others in the lab. I calculated the volumetric flow rates for one of the peristaltic pumps used in the lab for the flow circuit in cell-culture. The setting # is the bar # on the peristaltic pump controller (the 10 bars…