First, a quick publication update. Recently we’ve published a paper on our efforts towards computationally analyzing polymorphonuclear leukocyte (PMN) trafficking on a vascular endothelium via hand-built µSiM flow cell devices. We record videos in phase contrast with a 40x objective…
Category: NRG
Progress in Modelling Sepsis: Diminishing PMN chemotaxis towards fMLP gradients in µSiM Devices
Posted in NRG
m-µSiM platform: TEER and Permeability for Open-Well iPSC-ECs and viability test for future flow module
Introduction Tissue barriers regulate the absorption of nutrients, maintain homeostasis, and prevent toxins from entering. As previously introduced, trans-epithelial/endothelial electrical resistance (TEER) measurements offer a non-destructive and real-time method to quantitatively evaluate the integrity of tissue barriers. During the process,…
Posted in NRG
Biological Statistics SuperPlots Paper
For everyone conducting biological studies, this paper highlights how to think about sample size and replicates. It suggests using “SuperPlots”, which color codes data points from different experiments to better visualize the type of variability seen (i.e. random variability, experimental…
Posted in NRG
Localizing Basement Membrane Production in Coculture BBB Model
Introduction The basement membrane (BM) is a critical component of a healthy blood-brain barrier (BBB) model. Further, the BM breaks down in several diseases. Both pericytes and endothelial cells can contribute to basement membrane production. This post will explore a…
To Pulldown or Not to Pulldown, That is the Question: Testing the Utility of a Pulldown in µSiM-CA Bacteria Assays
The idea of adding a pulldown modification to our bacteria assay protocol came about after we started to believe that pore discovery was our rate limiting step of the transmission process, discussed here. Initially, it seemed to benefit our assay…
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Less is More: Lower Initial Bacteria OD Increases Transmission in uSiM-CA Bacteria Assays
On our quest to optimize bacteria assays using µSiM-CAs, we recently discovered that higher initial bacterial ODs decrease transmission in positive controls, described here. This was something we did not initially anticipate, engendering a new hypothesis around the idea that…
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A Serendipitous Disaster of Interesting Proportions: High OD Staphylococcus aureus Pulldown on 0.5 µm Microporous Membranes
After titrating our pulldown volume to avoid pulling Staphylococcus aureus through our membranes, discussed here, we found that 10 µL withdrawals allow us to continue to use the pulldown modification without compromising our results by providing false positive transmissions. With…
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The Big Squeeze: Unintentionally Pulling Staphylococcus aureus Through 0.5 µm Microporous Membranes
On our quest to improve the performance of bacterial assays that use the µSiM-CA by modifying our standardized protocol, our most recent modification has caused controversy. Described here, we recently decided to add a step to our protocol where we…
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Check Yourself Before You Wreck Yourself: Quality Control of 0.5 µm Microporous Membranes
For the osteomyelitis project which I have been working on for quite some time now, my collaborators and I have been working solely with 0.5 µm microporous membranes. To ensure that every membrane we work with has pores of the…
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Ghosts of Christmas Past: Live Imaging Bead Capture on 0.5 μm Microporous Membranes
The last time I did live imaging on the confocal microscope was in December of 2019. Then I was imaging the capture of 1.24 µm diameter beads in 1 µm slit pore membranes to visualize the difference between steric and…
Posted in NRG
