Ali Twelvetrees
@dozenoaks
Sir Henry Dale Fellow | Sheffield | cell & molecular neurobiology | kinesin, dynein, slow transport and neurodegeneration | preferably one molecule at a time | my opinions | she/her
Delighted to see the latest preprint progress to publication with Advanced Science. We built the protein FRET ruler as a bridge between biophysicists and cell biologists - realising the potential of FRET link measurements from molecules to cells. advanced.onlinelibrary.wiley.com/doi/10.1002/...
Although there are hundreds of mutations across the microtubule transport system linked disease, we focus here on the kinesin-1, kinesin-3 and cytoplasmic dynein families. These are the specialists in long distance transport that neurons rely on. onlinelibrary.wiley.com/doi/full/10....
Our latest review is out. We take a deep dive into the human mutations in microtubule motors that drive devastating neurodevelopmental and neurodegenerative diseases. Taking in motor protein biophysics, animal models of disease as well as patient phenotypes. onlinelibrary.wiley.com/doi/full/10....
Sunday. Third trimester. I have a mountain of work to get through and there is not one chair in the house that’s comfortable. Meanwhile my passenger is pulling yoga moves inside me.
Finally, a GABA-A receptor question on University Challenge. Knew that PhD was good for something. Sadly only the dog around to witness my prowess and she’s asleep…
Save the date for #NeuroBioUK26 Do you like neurons? Maybe glia? Molecules? Cell biology? - The venue is booked and we are excited to see you and your incredible science there on the 11th of September! Please do join our email list to stay up to date. neurobiouk.sites.sheffield.ac.uk
Most excitingly, it also works in cells - opening up the possibility of interpolating FRET read outs between single molecule studies and protein conformations in cell based ensemble assays (FLIM shown here). 3/
It works as either purified protein or measured directly in cell lysate for single molecule work. FRET changes are regular and predictable, and the proteins are 'well behaved' in all the settings we've tried so far. 2/
Fancy a fresh preprint for Friday? When we were first getting involved with single molecule FRET, there weren't any standard protein molecules that suited our applications to help us develop our pipeline. So we built some! A universal protein ladder for FRET. 🧵 1/
Doggo was making her feelings known about Sunday morning grant writing… I put the laptop down to wrestle her off and the cat has now taken up the defence. Sigh.
Poster boards are ready and waiting for #NeuroBioUK - let’s get this science party started 🥳
Cooling off after a hot day, Sheffield style. Here’s to the weekend! Don’t forget to get those abstracts in for #neurobiouk25 :) neurobiouk.sites.sheffield.ac.uk/registration
Friday gave us a double treat at our Single Molecules seminars. Fab talks from @annestraube.bsky.social and ECR speaker Dylan. Between them they covered AFM, optical trapping and TIRF assays - serving fascinating insights into photosynthesis and microtubule transport. Single molecules FTW!
Registration is now open for NeuroBioUK 2025! This year we're looking forward to hearing from our plenary speaker @cathyabbott.bsky.social. As ever, all other talks chosen from submitted abstracts so get submitting - we can't wait to see you there! neurobiouk.sites.sheffield.ac.uk/registration
A recent selection of found cats for #caturday, separated by a few hundred years and a few thousand miles…
Fantastic talk from @mathewhhorrocks.bsky.social for our Single Molecules @ Sheffield seminar series. Really creative applications of single molecule confocal systems for neurodegenerative disease research. Safe travels back to Edinburgh :) Thanks again to our sponsors @biochemsoc.bsky.social
Has anyone mentioned to the PM that a healthy life sciences sector is critically dependent on a healthy university system? One that includes humanities and arts that can deal properly in ethics and human experience? Taken from Starmer’s speech today -
Feeling totally shattered tonight after the presentation of a lifetime - my 10 year old’s year group. Kiddo is proud of me and not embarrassed of her mum (!!). The classes asked amazing questions - the true test that anyone followed anything I said. And now I am ready for bed!
Good morning from the sleepy bees at Sheffield Botanical Garden. Guess it must be spring?
Do you need some science joy in your life? Do you like neurons? Maybe glia? Molecules? Cell biology? Save the date for #NeuroBioUK25 - the venue is booked and we are excited to see you and your incredible science there on the 12th of September! neurobiouk.sites.sheffield.ac.uk
I could spend today in an existential crisis about the future of the world, or I could science… 🔬
With all the optimisation we got great reproducible smFRET data. If you’ve got access to one of these instruments and a regular TC facility, then all you need is a couple of wells in a 6 well plate and you can have enough protein to test out what’s going on in a sensitive biophysical assay! 7/10
Evie then optimised labelling time and concentration of ligands in our HEK cell expression system using in-gel fluorescence. CLIP is known to have much slower labelling kinetics than SNAP, but with this careful optimisation Evie got past this hurdle. 6/10
Confocal smFRET is very sensitive to contaminations, because you only want one fluorescent molecule in the confocal volume at a time. Evie used flow cytometry to optimise the wash out time of SNAP and CLIP ligands to minimise contamination from unbound fluorophores. 5/10
We went for self labelling enzymes CLIP and SNAP because they are small and give us access to stable fluorophores. Small tags are best for FRET to maximise the dynamic range of the assay. Self labelling enzymes avoid having to deal with issues from fluorescent protein maturation. 4/10