Monday, November 25, 2019

The Okinawa Analytical Instrument Network Meeting 2019!

Have you ever heard of OIST? I'll be honest, neither had I. I am, however, 100% confident that you will hear lots about it soon. OIST stands for the Okinawa Institute of Science and Technology and it might be the most beautiful campus on earth. No picture I've taken has done it justice at all. The facility is built on mountaintops to responsibly protect the ecosystems around it. And it is growing! 

I was inexplicably invited to talk at the Analytical Instrument Network Meeting this year along with some serious mass spectrometry experts. I'm going to ramble through an overview of this meeting here:

Day 1: Meet people, see the amazing facility and Q/A session.

This facility has an amazing array of instrumentation and hardware. From robotics through NMR and various arrays of GCMS and LCMS to support the needs of a facility dedicated to primary scientific discovery. The work they are doing is on a lot of organisms I've never even heard of. The best part of day #1, however, was the Q/A session. The speakers (who I'll detail below by talk on Day #2) and I were invited to sit down with the Instrument Analysis Section Staff (the people running the 40 analytical instruments!) and their collaborators who are utilizing the mass spectrometers the most.

This is a plea from the very bottom of my heart to everyone with an aging mass spectrometer: Please please do not retire that 10 year old Orbitrap. Don't trade it in for a $15,000 credit unless you really truly can't afford it with your new instrument. Do this instead: Set that instrument aside and let the power users of your facility -- or better yet -- students at your or nearby university's use it. Have a workshop and show people how to do the basic stuff, set it up with some nice generic workflow and let them go. It might be the best investment you've EVER made in the future.

OIST has done something similar. They have several LTQ Orbitraps and one is user walk-up. I know your objections. "The students will just break it!" Sure. Set it up so it's hard to. Capillary flow or, heck, analytical flow. Put a good desalting/trap system on it. Use a wider bore emitter. Give them enough training that they will mess it up some. The payout is astronomical.

I mean no offense to any biologists I've worked with in the past, present, or future but if your collaborator's best understanding of what happens after they give you the sample is something akin to this..... are handicapping yourself, them, and science in general.

They aren't going to ask the right questions of you or the technology. They are going to ask questions that LCMS is not the right solution for and the project will be a failure. Or they are going to ask questions that the technology could easily answer without knowing that while doing so they could learn much much more. (And how much longer are they going to even be interested in what we do?) Without some fundamental understanding of what we're doing in the lab (and to be fair, I do a decent amount of dancing in the lab when no one is around, and I KNOW I'm not the only one -- what else do you do when you're sonicating or centrifuging for 3 minutes?) we aren't speaking the same language. OIST previously had an ion trap that some users were trained to utilize and now they have an Orbitrap Classic. And you know what?

The questions the panelists and I received were some of the best I've EVER heard from a group of people consistently. I'm not kidding -- the biologists who are using the center resources know enough about what the instrument is doing that they are pushing the boundaries of what I or the other panelists (and I think we were talking about 75 years of mass spec experience on the panel, all told) knew or had thought about. I frantically started taking notes, because there were things the biologists cared about that we could absolutely be helping them with, that we aren't because we aren't thinking in the same terms. (Now, it probably doesn't hurt that the proteomics lead here, Alejandro Villar, might be a legitimate genius, but the end users here are impressive. And, yeah, there is probably an abundance of talent, but my bet is that most facility's could come close to replicate this model with similar success.)

As an aside -- this group does have what might be the solution for the hardest part of single cell analysis. They have a slow, methodical robot that is designed 100% toward reproducibility and accurate sample handling. It might cost more than an Eclipse, but how many single cell proteomes do you have to mess up before that seems negligible? Depends on the model, I'd guess!

Day 2: The actual meeting.

Our host Dr. Kazuo Yamauchi kicked off the workshop by reminding everyone that this was an interactive event (yeah! that's how you learn this stuff, unless you're a giant sample prep robot..)


Dr. Andreas Huhmer provided a remarkable optimization of single cell proteomics by his team beginning with label free analysis (<500 proteins) to ScoPE-MS (~1,000 proteins) through adding FAIMS and then Direct Search (the Eclipse function that presearchest TMT data to make sure it's decent before doing SPS MS3.

Huge takeaway here (and he's sent me a reference) if you are doing ScoPE-MS you do not want to overload your carrier channel!! It seems like you could just load more and more peptides into your carrier and you will get more data. You will get more peptide IDs -- sure -- but you will screw up your reporter ion signals. You'll suppress the single cells. To be honest -- I was sure I was ruining my sample prep when did ScoPE-MS (and I probably didn't do it any favors) but I did think that loading more carrier was a great next step when the first experiment didn't look great -- and -- I think my data looks like what he was describing. More on this later, for sure.

Professor Yet-Ran Chen -- studies of plant immunology leading to a massive success story toward leveraging plant immune function to getting the world away from pesticides with endogenous immune peptides. Check out this new paper on the topic. Also -- his team has developed a method for using Mascot to search for endogenous peptides. He's doing this with plant immune peptides -- strategies to steal for HLA/MHCs? Probably! (I mean...I was just impressed to learn that plants had immune systems, to be honest)

Professor Newman Siu Kwan -- His group does a lot of work with proteomics and PTMs and aging, and that is what he spoke about. His Google Scholar indicates that a lot of stuff deals with...compounds that mediate...age related terribleness. 100% worth searching through this!

Professor Takeshi Bamba -- Metabolomics by supercritical fluids!  This talk began with me frantically taking notes and looking up what supercritical fluids are. His team uses pressurezed CO2 to separate metabolites! You can separate lipid classes with it. And it can be coupled to HPLC and then to mass spec and he's partnered with vendors to bring this technology to market. I'm still not 100% sure I get it, but it's worth checking out.

Other highlights were great questions and conversations and the pages of notes that I have.

I'm still not sure how I got included in this, but I'm deeply greatful to the organizers for this experience and have made some friends here that I hope I have for life. I even got to stick around and sit in front of their instruments and learn and bounce ideas back and forth with this amazing team.

OIST has a dream of doing something truly special to improve the environment and science of the world and they are well on their way to accomplishing it on this ridiculously beautiful and fun little island!

Edit: Right as I was boarding the next leg of my 20+ hour trip home I received the picture at the top from Dr. Yamauchi. I made the joke repeatedly that all you would see was the projector bouncing light off my shiny head. I am waiting for my first of many planes today to board and I can't stop laughing because I was more right than I knew. A consequence of the great aerodynamics I get these days!

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