Two weeks ago (as of the time of writing this post), I wrote a post on a dead end filtration experiment that I ran in order to demonstrate particle capture in pores, modeling a simple, purified exosome capture system that we could use for imaging or analysis. However, when I ran that experiment, I done…
This is the second in my series of posts that will be on my model exosome system. As I am trying to define the appropriate conditions for separating exosomes from blood and other fluids, I am first going through and trying to isolate each important parameter and see how they affect the capture of these…
In trying to determine the best conditions for capturing exosomes, one of the important questions that we have is what produces the case where exosomes are caught directly in the pores, not on top, but flush with the surface. This should be the size where the particles are almost exactly the size of the pores,…
A few weeks ago, Josh and JP brought me some 3 window microslit membranes that had edge aligned 8 and 9 μm slits. While these may seem to be quite large for most applications, they are in fact perfect for working with whole blood. The experiments that we have planned for these membranes are based on the…
This is my third post that is related to the NanoAuger system that I had the opportunity to use while I was home over break. However, instead of quantum dots and exosomes, this time I will be talking about data pertaining to Greg’s magnesium fluoride membranes that are used for Raman imaging. And since I…
Alright, so this is kind of a big update and I will probably split it into a couple of parts. Basically, in a search for a way of confirming that the particles we are seeing on the membranes, I have tried several techniques with varying degrees of success. The initial experiment involved me conjugating an…
Since I am posting data that is acquired using Auger electron spectroscopy, I thought that it might be a good idea to write a post about the theory of the instrument, to better understand why this data is useful and relevant. The Auger effect is a phenomenon in which an atom that is excited by…
In attempting to generate control systems for our exosome work as well as to potentially develop a system that would help some collaborators at the University of Illinois, I have been working with the ultracentrifuge again to hopefully isolate a pure fraction of exosomes, devoid of any protein components. This would allow us to start…
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 TEM grid (we were able to cut it out and weld it to a grid…
As a step to identifying the particles that I am capturing in my systems, I am attempting to do an immunogold labeling with a CD 63 primary antibody and a 10 nm gold conjugated secondary. It is possible to run this analysis in an SEM, but the best confirmation method is using a TEM and…