I’ve been gathering data for a paper using my microporous MgF2 nanomembranes so here are some sample figures for such a paper.
Herein, we are trying to create a porous, Raman-compatible substrate for use in cell culture. Researchers have used Raman microspectroscopy to perform quantitative non-invasive measurements of biomolecules in cells. It’s a very exciting technique. While silicon based substrates have excellent properties for cell culture, they have strong Raman background signals and absorb a lot of laser power at the illumination wavelengths we would like to use. MgF2, Quartz, and CaF2 are considered Raman-compatible for their substantially weaker backgrounds. Researchers currently deposit or culture cells on non-permeable coverglasses made of these materials. However, we have known that cells sometimes require a permeable substrate to adopt morphologies that are more consistent with in vivo studies of morphology. So using our microporous nitride substrates, we wish to create a substrate that will have both the permeability and the Raman-compatibility necessary to facilitate more accurate Raman studies of those types of cells.
Figure 1. MgF2 nanomembrane material properties. (a)-(b) MgF2 relief pattern transfer, with cartoon crossections of microporous films (not to scale). Beginning with a freestanding film of microporous silicon nitride (a), MgF2 is evaporated onto the substrate (200 nm, 0.1-0.3 nm/sec, 250 °C, Platen Rotation), coating the porous substrate, resulting in a hybrid material. (b) The substrate is then inverted and purified using RIE (90% CHF3, 10% Oxygen, 75 mTorr, 100 W), releasing a freestanding nanoporous film of MgF2. (c) A SEM image normal to the membrane plane shows the infilling effect of the direct evaporation process, as the template pores narrow from their designed 500 nm diameters to approximately 325 nm. (d) SEM crossection of a partially etched MgF2 nanomembrane (with some template backing remaining), showing film thickness close to the targeted 200 nm.
EDS measurement of the composition of the freestanding membrane. While the Mg and F signals are preserved across the freestanding region, the Si signal is not. The membranes are >85% MgF2 in the freestanding region. As a side note, there was some sample drift, so the image is blurred. This is a 30kX image.
Before Etch
After Etch
Previous Figure with 50 nm thick nanoporous MgF2. Here the burst pressure for the 200 nm microporous MgF2 is about 7 PSI, over a 5-slot structure instead of a square window.
20X Live/Dead stain image of P6 HUVECS (Day 7) on fabricated microporous MgF2 grown in MCDB131 media. Eventually, we will use ARPE-19 Cells with tight junction staining which have a characteristic response to permeable substrates.
We will have to reestabilish the Raman background of the membranes, but the material composition looks to be the same as with my nanoporous material. HUVECS seem to stick ok without using any additional coatings.
My goal for this experiment was to see whether vacuoles form in primary cells. If they did, this makes the case that the results with bEnd3 are more physiologically relevant since primary cells are better models of in vivo behavior. Here, I simply seeded P8 HUVEC at 50000 cells/cm2 on plastic wells, PET and pnc-Si…
Attached is a paper I’d like to review on Thursday (June 11, 2015). This group is using two planar surfaces in an attempt to mimic a 3D cell culture environment. This is similar to our claims that cells growing on a permeable membrane can be akin to a slice of a 3D matrix. I met…
In support of the bone infection studies done in the Schwarz lab at URMC, I have recently been tasked with coordinating a new collaboration with Dr. Yates lab in the Chemical Engineering department. The goal of this collaboration is to establish a method to coat our membranes with hydroxyapatite crystals. Two different methods were attempted,…
Wafer 403 was processed last week using the new thermal oxide/TEOS steps outlined in the previous posts. I took images from six positions at different radial positions on different axis. All six samples looked relatively uniform! (1,0) – 35kx (0,-2) – 35kx (3,0) – 35kx (4,0) – 35kx (0,5) – 35kx We know that the…
Here is the full updated Adsorption Quantification figure: We were concerned that cellulose appeared to adsorb more than PES when the literature clearly states that it shouldn’t due to its hydrophilicity. We considered that this same hydrophilicity might be part of the problem with our method. Macroscopically, it appears that the protein solution applied to…
I recently performed an experimental separation with 4 proteins in 100 and 10 mM KCl. All proteins are negatively charged at the pH of the experiments, and we would expect that less would pass through the membrane in the 10 mM KCl solution. Here’s the gel: You can see that the beta-gal in particular does…