Not too long after publishing version 1 of the SmartFlare paper at Science Open, we received a couple of very helpful reviews. We’ve been working on the points made by the reviewers to improve the manuscript for version 2 (coming very shortly!). Below are some thoughts on a couple of the comments and a quick update on some new data in the works.
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.
I recently discussed the SmartFlare project at a joint Sée and Lévy Group Meeting, partially extolling the virtues of Open Science but mostly focussing on the data acquired so far and some of the proposed experiments.
Regarding the SmartFlare project, the three big questions that formed the basis of the talk were:
- Why do only some cells take up SmartFlares?
- Where are the SmartFlares?
- Why are the VEGF and Scrambled SmartFlares fluorescent in puncta?
Below are some of the interesting points that were made during the meeting. Replies at the time (as best as I can remember them) in green:
With the Undergraduate project reports now marked, Gemma has kindly agreed to post her project report on the blog. You can find a pdf of the report here.
As most of the methods and results are already on the blog, a slightly abridged and edited version of the discussion section is replicated below:
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).