Insights from industry

Microscopy & High-Speed Imaging

Professor Mervyn Miles met with James Vicary for the NuNano blog, to discuss where his interest in high-speed imaging and microscopy came from and also to examine new emerging research.

How did you become involved in microscopy?

I never set out to work with microscopes. My Ph.D. actually focused on understanding polymer samples and for the first part of my career, understanding and predicting the structure of polymer samples remained my main point of study.

During my time at Birmingham University, I was granted access to a million electron volt (MeV) electron microscope. Now in the normal course of things polymer samples tended to get destroyed after a couple of seconds in electron microscopes operating in the 100 keV range. Not with this one though! Using the mega-volt microscope, I could see all sorts of pretty diffraction patterns.

From that point on I was hooked on microscopy and imaging - though at that stage only because it was a fascinating tool to facilitate my polymer research.

Would you say that a vital part of the polymer research you were involved in related to finding the best possible imaging equipment?

Yes, you could say that. I went to Germany to do my post-doc in synthetic polymer physics studying under Herbert Gleiter[1]. He had a whole new way of looking at polymers, again using electron microscopes[2]. During this time I obtained some pretty interesting images, showing the nanoscale structures of polymer fibers produced under elongational flow stretched single molecules[3].

After I completed my post-doc in Germany I went to Case Western Reserve University, Cleveland, Ohio, which at the time had the best polymer department in the U.S. After just one year in the U.S., I started my first period in Bristol.

My next move, still focusing on polymer research, was to Norwich, to work – rather bizarrely – for the Institute of Food Research. They had loads of money to do basic science which in turn meant we weren’t constrained in the science we could do. Though ideally, the sample should be edible!

Our head of division was a fascinating, enigmatic guy called Henry Chan. He was the person who first introduced me to and encouraged me to work with the new science of scanning tunneling microscopy (STM). Nobody really knew what it was about of course. Even at the European Bioscience Physical Congress, held in Bristol in 1984 – a huge conference with many parallel sessions including one on x-ray microscopy.

Through Dr. Chan’s and Dr. Morris’s encouragement, I took one of the protein molecules I had been studying with small-angle x-ray scattering (SAXS) and deposited on an x-ray mirror’s surface, amorphous carbon, and looked at it via the STM with Mark Welland. Through the images we obtained, we discovered the 3D structure the STM was showing us of individual molecules was as I had predicted from SAXS - we got one of the first pictures of a single protein molecule[4].

At what point did you start getting involved in instrumentation development?

By this point, I was increasingly interested in the latest and best developments in microscopy. My focus was on improving the substrate for immobilizing biomolecules rather than the tool itself though. I was always looking for the best substrate materials.

In 1989 I moved back to the University of Bristol. I was supposed to be working on x-ray diffraction and scattering, but clearly, STM was going to be huge, so I applied for an STM grant immediately... and we were awarded it in April 1990. This meant we finally had our own STM to play with!

Around this time one of my undergraduate project students built a scanning near-field optical microscope (SNOM) which achieved the best resolution of anywhere in the world and this is really where developing instruments came in for me, from around 1990.

Shortly after that, I put in for a grant and got an Atomic Force Microscope (AFM). I recruited a post-doc, Terry McMaster, from Norwich, and we began work on AFM of biomolecules.

Where did the idea for setting up NuNano come from?

Frustrated by the varying quality of existing AFM probes, we thought maybe there was an opportunity to make improvements. It was the brain child of my colleague and co-founder Heinrich Hoerber. The mix of what seemed to me to be a great idea with the enthusiasm and drive of former Ph.D. student and post-doc James Vicary, made setting up the company a no-brainer.

James has done a brilliant job of implementing the idea and turning it into a successful product. And, importantly developing out from that original idea to produce the kind of game-changing probes that no-one else is working on yet, such as the ultra-soft high-speed vertically-oriented probes (VOPs).

Is there a new area of research you are interested in and what do you think is exciting about the future of nanotechnology?

I think the use of high-speed VOP force microscopy in true non-contact mode, with zero normal force, on living cells will make a major impact. The ability to see signaling, transport and whole changes in structure at the cell membrane will give exciting new information.

Using the high-speed vertical probes for example, as I have done lately with work I’ve been doing around Alzheimer’s, just makes you realize how much more there still is to be explored in the world of force microscopy.

Imaging membranes at high speed means you don’t end up making holes in the lipid (and thus destroying the sample). We’ve produced the most amazing images of membrane samples where you can actually see the Amyloid proteins destroying the membranes.

It's exciting and important work, and we're looking at ways to go beyond just imaging.

High-speed (left) and normal speed (right) AFM images of the same area of a model multi-lipid component neuronal membrane. The contrast corresponds to the slightly different height of each lipid.

High-speed (left) and normal speed (right) AFM images of the same area of a model multi-lipid component neuronal membrane. The contrast corresponds to the slightly different height of each lipid.
(Image credit: Morgan Robinson/Zoya Leonenko/Loren Picco/Ravi Sharma/Mervyn Miles)

[1] H Gleiter, “Nanoglasses: A new kind of Noncrystalline Material and Way to an Age of New Technologies?” Small 12 (2016) 2225–2233
<[2] J Petermann and H Gleiter, “Direct Observation of Amorphous And Crystalline Regions in Polymers by Defocus Imaging”,Philosophical Magazine 31 (1975) 929-934
[3] MJ Miles,  J Petermann & H Gleiter, “Deformation Mechanism of ‘Hard’ Elastic Fibres’, Colloid & Polymer Science, 62 (1977) 6-8.
[4] ME Welland et al., "The Structure of the Globular Protein Vicilin revealed by Scanning Tunnelling Microscopy", International Journal of Biological Macromolecules, 11 (1989) 29-32.

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