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More on Fluorescent Microscopy Chemistry Prizes

I wanted to note (with surprise!) that one of this year’s Nobel laureates actually showed up in the comments section of the post I wrote about him. You’d think his schedule would be busier at the moment (!), but here’s what he had to say:

A friend pointed this site/thread out to me. I apologize if I was unclear in the interview. #3 and #32 have it right — I have too much respect for you guys, and don’t deserve to be considered a chemist. My field is entirely dependent upon your good works, and I suspect I’ll be personally more dependent upon your work as I age.
Cheers, Eric Betzig

And it’s for sure that most of the readers around here are not physicists nor optical engineers, too! I think science is too important for food fights about whose part of it is where – we’re all working on Francis Bacon’s program of “the effecting of all things possible”, and there’s plenty for everyone to do. Thanks very much to Betzig for taking the time to leave the clarification.
rhodamine
Bacterial probe
With that in mind, I was looking this morning at the various tabs I have open on my browser for blogging subjects, and noticed that one of them (from a week or so back) was a paper on super-resolution fluorescent probes. And it’s from one of the other chemistry Nobel winners this year, William Moerner at Stanford! Shown is the rhodamine structure that they’re using, which can switch from a nonfluorescent state to a highly fluorescent one. Moerner and his collaborators at Kent State investigated a series of substituted variants of this scaffold, and found one that seems to be nontoxic, very capable of surface labeling of bacterial cells, and is photoswitchable at a convenient wavelength. (Many other photoswitchable probes need UV wavelengths to work, which bacteria understandably don’t care for very much).
Shown below the structure drawing is an example of the resolution this probe can provide, using Moerner’s double-helix point-spread-function, which despite its name is not an elaborate football betting scheme. That’s a single cell of Caulobacter crescentus, and you can see that the dye is almost entirely localized on the cell surface, and that ridiculously high resolutions can be obtained. Being able to resolve features inside and around bacterial cells is going to be very interesting in antibiotic development, and this is the kind of work that’s making it possible.
Oh, and just a note: this is a JACS paper. A chemistry Nobel laureate’s most recent paper shows up in a chemistry journal – that should make people happy!

8 comments on “More on Fluorescent Microscopy Chemistry Prizes”

  1. Anonymous says:

    “That’s a single cell of Caulobacter crescentus, and you can see that the dye is almost entirely localized on the cell surface”
    Damn, for a second there I was thinking (hoping) that was a picture showing the kink in just one fluorescent molecule.

  2. Twelve says:

    Hi Derek – that’s a very graceful note, and it certainly could be from Betzig, but of course it’s trivial to sign yourself into this system with any name you choose. Did you hear from another direction that this is the real McCoy?

  3. Derek Lowe says:

    #2 Twelve –
    In identity-checking situations like this, I go to the IP that left the comment. This one was from the Howard Hughes labs in Virginia, as it should be. (I had the same thoughts!)

  4. The Iron Chemist says:

    Regarding Moerner: it’s a bit odd to hear everyone call him “William.” I believe he still goes by “W. E.”

  5. Anonymous says:

    Let me preface by telling that I have great respect for the Nobel winners who I’ve had the opportunity to listen to in so many lectures and conferences. My lab has also been working on superresolution microscopy and the problems and the elegant solutions these people have come up with are indeed commendable. 
    However, I feel like this Nobel announcement is premature. For a decade-old method, you would see by typing superresolution in pubmed that there are no more than actual “biology” papers that use this method in proper biological settings. Its still chemists and optics people publishing “better” methods and reagents, but something is stopping the biologists from adopting the methods. What is that? Here’s a list:
    1. It’s complicated. Of course superresolution is complicated, so no lab can just take a microscope and  image it (except for structured illumination, which “only” gives a marginal resolution increase but can be easily configured as a core-facility setup that anyone can switch on and use). 
    2. Sample preparation kills everything. This is the bigger issue that practically screws up superresolution: fixing the sample more often than not destroys sample at a larger distance scale than these resolution methods can image at. So every time we tried doing STORM what we got was images of single molecules which we were sure we were real, but they were just spread around everywhere. That’s why you will see most publications on superresolution will try to show you beautiful images of actin and tubulin, but nothing else. The other things are that much harder to fix to a defined structure. Betzig et al’s lab is definitely working somewhat in the right direction trying to get structured illumination (which I think is the only practically usable superresolution method) in more biologically relevant projects. But we’re still a long way to go before this method becomes actually useful. Unfortunately because of this premature award, any scientist who actually solves this real problem in the future might be denied even a chance at this prize. 
    3. EM can still do better. Even after the advent of superresolution in 2006/7, you still see more biology papers today with EM images than superresolution images. That’s because EM still gives a much higher resolution. Of course you can’t see your proteins, but immunogold (as tricky as it is, its still less tricky than superresolution IMO) and Tsen’s MiniSOG are ways that can circumvent that problem. And with good sample prep you can see the membrane bilayer. No superres method can still do that.
    4. You still can’t do proper live superresolution: Except for studies of lipid rafts (which apparently will never go away) there has been no real application of superresolution in live cells that was not just there for demonstration purposes. Gustaffson’s Structured illumination does help that but the Nobel was not given for structured illumination (Though to note that Gustaffson passed away unexpectedly in 2011, and if he was around, would have probably shared this prize).
    With so many unanswered questions, why did they give the prize already? In my opinion, superresolution is the microarray of the 2000’s. Amazing on paper, people push out a large amount of data, but eventually everyone realizes most of that data is useless because of unsolvable problems. Sure microarrays paved way for other interesting methods like ChIP and maybe even RNAseq, but imagine if the Nobel committee had awarded the prize for microarrays before these methods even came to the light.

  6. Anonymous says:

    #5
    Interesting view and perspective. Does anyone who works in this area have rebuttals to these statements?

  7. naoh3K says:

    #5, #6
    I have done considerable work designing and synthesizing photoactivatable materials for imaging. I have never got to work in the side of the field where the probes are applied in order to obtain images, so if what I am about to say sounds naive, please forgive me.
    From my perspective, it seems like the biggest hurdle to the widespread usage of SR imaging could simply be the exotic nature of the probes required to do the imaging. Packing functionalities into one molecule that makes it targeted, photoactivatable, soluble, non-toxic, and operate at proper wavelength (like in the following link: http://pubs.acs.org/doi/abs/10.1021/ja1044192) is doable but makes commercialization impractical. I guess my point is that if there were more readily available probes that possessed a lot of these attributes and were affordable to make, then the use of SR imaging could be more widespread. But as it stands, when you want to do this type of imaging, it seems you have to collaborate with an institution that will handle the grunt work of synthesizing new materials. Once proper fluorescent probes are readily available, we can get into applications that are for more than just demonstration purposes.
    I certainly understand how some might call the awarding of this Nobel premature. But does the work have to immediately lead to something that any common technician can pull off? Or can we appreciate that finding a workaround for the diffraction limit is amazing in its own right?

  8. Project Osprey says:

    A bit a tangent this but a highly entertaining one. Here’s what happens which you try to take a Nobel prize through airport security.
    http://blogs.scientificamerican.com/observations/2014/10/10/nobel-prize-airport-security/

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