Used Thermofisher SiteClick qdot antibody labeling kit 625.  Began with 4.0mg/mL IgG and conjugated it with an unknown amount of qDots, methods provided by the SiteClick kit. The final stage in the manual [1] is an optional cleaning of the…

I am trying to understand the development of endothelial barrier properties as the cells remodel under the influence of physiological shear stresses. I culture HUVECs inside my device on 100 micron thin type I collagen gel. I coat the gel…

First off, this is the protocol for the staining and analyzing the data is shown below and was used for all of the YAP experiments: For the substrates, we wanted to use 0.5μm HP, 0.5μm LP, 3μm HP, TCP, and nonporous…

Goal: study HUVEC migration on different porous substrates. HP = high porosity 0.5 μm SiO2 membrane LP = low porosity 0.5 μm SiO2 membrane NP = non-porous SiO2 membrane TCP = tissue culture polystyrene (~3 GPa [ref 1, 2]) P184 = sylgard 184…

To follow up on my recent posts about HUVEC YAP localization after 24 and 72 hours (https://trace-bmps.org/blog/data/2016/10/19/yaptaz-passage-one-staining and https://trace-bmps.org/blog/data/2016/10/11/yaptaz-staining-on-membranes-and-nonporous-sio2) , we conducted the same study using Adipose Derived Stem Cells.  Due to the fact that these are mesenchymal stem cells,…

(This post is not finished, but my flight keeps getting delayed in 20 minute increments, so better to publish it unfinished than to not get it up before NRG in case I’m not able to make it. Please forgive typos…

This is pretty much an update to a previous post on trying to identify exosomes using an immunogold staining procedure, FIB lift-out and the TEM. After multiple attempts to cut out a section of the membrane for attachment to a…

Hello Everyone, Following my experiments with Track Etched Membranes: So we came up with this idea to perform SEM imaging. Following are the images that i took for my experiment with 100 nm beads over 100 nm pores after releasing…

There are two longitudinal studies for Chronic Kidney Disease (CKD). Ancillary studies can be set up to share the collected data, bio samples, or even request additional data to be collected. These Ancillary studies can be funded through normal channels,…

Purpose: Study the hemocompatibility of different substrates using platelet activation as the indicator.   Extraction of platelet rich plasma (PRP): PRP were prepared from whole blood following the standard protocol outlined by Abcam (http://www.abcam.com/protocols/isolation-of-human-platelets-from-whole-blood). To describe briefly, 9 mL whole blood…