Following on from the last post, the Uptake Control SmartFlares were loaded into cells, prepared and taken to the Electron Microscope.
Throughout all of the fluorescent microscopy experiments, we’ve been seeing SmartFlare fluorescence in punctate structures. Using these techniques, it’s very difficult to say with certainty that the SmartFlares are within vesicles or (for example) aggregated/agglomerated in the cytosol. As this is pertinent to the current investigation, we took cells loaded with SmartFlares to the electron microscope which has the resolution to discern membrane structures as well as the gold particles.
The Uptake Control SmartFlare was opened first to help us to establish a loading protocol. As such, the physical characterisation of that SmartFlare was performed in an earlier post. Thanks once again go to Joan Comenge for taking the VEGF and Scrambled control SmartFlare to the Transmission Electron Microscope (TEM).
As with any reagent, it’s important to know what you’re dealing with. The size of the gold nanoparticles at the centre of a SmartFlare will dictate the number of oligonucleotides that can be bound to the surface. Likewise, if the GNPs are aggregated, that will affect the way they interact with cells. In order to visualise the SmartFlares, lab member Joan Comenge took a sample of the uptake control to the Transmission Electron Microscope (TEM).