SmartFlare Paper Revisions

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.

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TEM of VEGF SmartFlares after uptake

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.

2015-07-14-TEM01 Continue reading

Discussion Points from SmartFlare Group Meeting

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.

By Andreas E. Neuhold[CC BY 3.0], via Wikimedia Commons
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:

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Imaging SmartFlares +/- DMOG (Live)

The purpose of these experiments is to be able to visualise the SmartFlares (SFs) in the cell after having followed the loading protocol provided by the manufacturers. After 16-20h the SFs should be in a position to interact with mRNA in the cell and this means a cytosolic localisation. We will use DMOG to stimulate VEGF mRNA production in the cell.

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