Yesterday I tried a gedankenexperiment via Twitter, and asked:
If you could visualise a protein relative to an intracellular structure/organelle at ~5 nm resolution, which one would you pick and why?
- Myosin Va and cargo on actin filaments in melanocytes – Cleidson Alves @cleidson_alves
- COPII components relative to ER and Golgi for export of big proteins – David Stephens @David_S_Bristol
- Actin inside an axon, AIS, shaft presynaptic bouton relative to membrane and vesicles – Christophe Leterrier @christlet
- Cargo/vesicle and motor, ideally with a co-reporter of motor activity – Ali Twelvetrees @dozenoaks
- Dynein on K-fibres. If it was a fixed view dynein on kinetochores, localisation relative to Ndc80 or Mad1 – Eric Griffis @DrGriff34
- See definitively if TACC3/ch-TOG is at the centrosome or not – Hadrien Mary @HadiM_
- Pericentriolar proteins relative to centrioles. And Arp2/3 and centrioles – Manuel Théry @ManuelTHERY
- Arp2/3 and centrioles was seconded by Alexandre Carisey @alexcarisey
- RhoGTPases near cell-cell contacts in endothelial cells. No good antibodies for this – Joachim Goedhart @joachimgoedhart
- Integrin and filopdia tips, what structures are formed there – Guillaume Jacquemet @guijacquemet
It’s a tough question because the simplest answer to “which protein” is the “the one I am most interested in” – I mean who wouldn’t want to see that at unprecedented resolution – but I was more interested in the “why” part. I’m conscious of the fact that breaking the resolution limit in light microscopy has not yielded many answers to outstanding questions so far.
OK, it was less a thought experiment and more like trying to crowd-source suggestions. We have some new technology that we’d like to put through its paces and apply to interesting cell biological questions. Thanks to everybody for their input.
If you want to make an additional suggestion, please leave a comment.
Edit 2016-03-13: Stéphane Vassilopoulos chipped in on Twitter. “dynamin 2 oligomers right on the actin cytoskeleton” he is @Biosdfp
The post title is taken from “What Can You See?” by The Seahorses off their unreleased follow up album to Do It Yourself, which may have been called Minus Blue.
2 thoughts on “What Can You See?”
Why? because centrioles are the typical structures which size (100x400nm) is too small to be seen in classical microscopy and big enough to see lots of interesting details with a 10 nm resolution. Nobody knows the exact contribution of such a highly regular structure within the amorphous cytoplasm. Looking with this resolution would be a great way to shine some light on this unexplained point which is certainly critical for cell physiology otherwise evolution would not have selected it so strongly.
Centrosome is the surprising combination of a highly controled and regular structure, the centrioles, in an amorphous dense cloud of clustered proteins, the pericentriolar material. Your imaging technique could tell us more about this unique coupling in cell organization.
Thanks for the comment Manuel. I like the idea of looking at PCM or at Arp2/3 a lot. The nice thing is that the centrosome is easy to find by co-ordinating light and electron microscopy, plus it has defined features for us to localise proteins to, distal end etc.
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