We previously explored cell migration and cell-substrate interactions in the form of altered f-actin polymerization, focal adhesion formation, and fibronectin fibrillogenesis on the discontinuous substrates (porous membranes and non-fouling PEG islands). Here, we were curious about mechano-sensing on these substrates. We first investigated the f-actin polymerization and focal adhesion formation of Adipose-derived stem cells (ADSCs)…

As a quick update on the ECM masking with Parylene membrane, we made changes to the device designs. Instead of relying on the peel-off process, we used a simplified design, with the membrane serving as the roof of the channels. Here is the latest design:   For these experiments we continued to use relatively low…

Now that I am working on labeling my 5637 EVs, I have finally gotten around to getting a good labeled sample and a good negative control. The samples were prepared as follows: 5637 EVs were thawed from a frozen stock and 1 μL was diluted into 1 mL of labeling fluid. The labeling fluid contained…

Introduction Sphingosine-1-phosphate: production and endothelial cell signaling Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid produced through the phosphorylation of sphingosine, a component of the plasma membrane, by SphK1/2. Many cells produce and store S1P, the largest source of which is platelets, due to their high native SphK activity. Erythrocytes and leukocytes are also sources of S1P…

Introduction EC Apicobasal polarity in immunity has and will continue to be a theme of my thesis. I have presented on bits and pieces of this work over the past year or so, but now that this work is under review and more or less complete pending reviewer comments, I wanted to summarize our finds…

Since precision printing FITC KODE molecules into the windows of our 3 slot chips I have begun testing both the uniformity and the functionality of surface coatings of these molecules. In pursuit of coating uniformity I have tested several different methods of surface coating to determine which method provides the greatest uniformity. In this same…

This post serves as an introduction to the uni-axial motion envisioned to be integrated into the tendon-on-chip platform. As discussed previously we are working in junction with Dr. Awad from the Center for Musculoskeletal Research (CMSR) center at UR to develop an in vitro platform to study fibrosis and inflammation in tendon tissue. One important…

Hey, this will be a rather short post, but kind of important for the current work on my 5637 EV capture. While working over the break, I was messing around with capturing CFSE labeled 5637 EVs and I noticed that there were many large clusters on the membrane surface (Figure 1A). This was concerning to…

Objective: Determine the minimum required area and number of exploratory cytoplasmic protrusions for sensing ECM alignment and causing cell polarization. Hypothesis: Cells only need to sense collagen alignment through exploratory protrusions extended during initial phases of cell seeding and not throughout the entirety of the cell, and a minimum number or size of protrusions is…