First time playing around with single molecule STORM imaging of PI3P on endosomes using recombinant biosensors. This was great fun with our new fellow Ruby Peters and I see lots of potential for this in the future!

I want to share one of my favourite #microscopy images with you all:
Epithelial cells in a monolayer (NMuMG) after undergoing EMT stained for N-Cadherin and actin show beautifully interdigitated cell junctions. Bonus points for spotting the actin asters ⭐

By segmenting the beads we can quantify these differences and show that active Arf6 strongly enhances the reaction speed of phosphoinositide conversion by the PIP5K. This has drastic effect on membrane trafficking pathways! www.cell.com/current-biol...

Importantly it also allows us to directly compare beads with different surface functionalisation. Here I attached the constitutively active mutant of the small GTPase Arf6 (Arf6-QL) and the inactive mutant to beads and we see complete separate reaction kinetics!

These realtime kinetics allow us to monitor individual reactions and revealed strong stochastic effects due to complex feedback loops as has been shown by Scott Hansen and Jay Groves www.pnas.org/doi/full/10....

In my first real post I want to show the key experiment that revealed to me what I want my own lab to be about:
The conversion of phosphoinositides!
By using fluorescent biosensors and membrane coated beads I was able to reconstitute the real time kinetics of PI(4)P to PI(4,5)P2 conversion by PIP5K

These realtime kinetics allow us to monitor individual reactions and revealed strong stochastic effects due to complex feedback loops as has been shown by Scott Hansen and Jay Groves previously www.pnas.org/doi/full/10....