Taking an atomic view of health is means understanding the tiniest scales and interactions of why molecules act one way or the other within a cell.
Professor David Punihaole at the University of Vermont leads a research team working with materials and medicines at that molecular scale, combining cutting edge spectroscopic techniques and microscopic views to investigate the fundamental chemistry behind health.
Read the original article: https://doi.org/10.1021/acs.jpcb.2c04415
Image Source: Adobe stock / Drazen
The following transcript is automatically generated
00:00:06 Will Mountford
Hello I’m will welcome to researchpod.
00:00:09 Will Mountford
Understanding the tiniest scales and interactions of why molecules act one way or the other within a cell is so far removed from a whole body view of health that when you hear phrases like nanoparticle drug delivery, you’d be forgiven for the shock of being reminded that everything, even you, it’s just atoms all the way down.
00:00:30 Will Mountford
Professor David Punihaole at the University of Vermont leads a research team working with materials and medicines at that molecular scale, combining cutting edge spectroscopic techniques and microscopic views to investigate and have action on the fundamental chemistry behind all our macroscale health.
00:00:51 Will Mountford
Doctor David pinhole hello.
00:00:53 Prof David Punihaole
Hi, how’s it going?
00:00:54 Will Mountford
I’m very well.
00:00:55 Will Mountford
Could you tell us a bit about yourself, your background and your research, and what brings us here today?
00:01:00 Prof David Punihaole
So my name is David Pony Holly and I am an assistant professor of chemistry at the University of Vermont. My research group is relatively new. I came to Vermont in August of 2020 during the height of COVID.
00:01:14 Prof David Punihaole
I’ve been setting up my lab since then.
00:01:16 Prof David Punihaole
My research group is interested in developing what we call chemical imaging.
00:01:24 Prof David Punihaole
Tools that can.
00:01:25 Prof David Punihaole
Be used to not only visualize the location of molecules for example, but also means to look at their structural dynamics and interactions to learn something about how these molecules behave chemically, which plays an important role in biological functions.
00:01:43 Prof David Punihaole
And this research interest really stems from my background. I received my my Bachelors of Science and Molecular biology.
00:01:51 Prof David Punihaole
And my PhD in molecular biophysics and structural biology, both from the University of Pittsburgh.
00:01:57 Prof David Punihaole
And as I said, my research interests stem from my background, really trying to understand the molecular behavior of proteins and DNA and other types of biological macromolecules.
00:02:11 Will Mountford
You say there’s anything I don’t know, philosophical, that you find in connecting.
00:02:16 Will Mountford
The whole human outcomes to these atomic interactions and atomic changes kind of the very different scales being personal when played out at.
00:02:25 Prof David Punihaole
Yeah. So I I think back to a quote from the great physicist and Nobel Prize winner Richard Feynman said that if you want to understand, you know, I’m going to paraphrase him. But he essentially said that if you want to understand.
00:02:39 Prof David Punihaole
Biology, be it the molecular basis for human diseases or just the fundamental workings of life, you have to recognize the fact that all biological molecules are composed of atoms, and that it’s the wiggling and jiggling of those atoms that give rise to the functions of proteins and.
00:02:59 Prof David Punihaole
DNA, RNA and all sorts of other types of biomolecule.
00:03:04 Prof David Punihaole
So I think trying to understand those wiggling and jiggling of those atoms is extremely important to to develop a holistic bottom up view of how life works.
00:03:23 Will Mountford
Let’s come back to the pinhole lab which you mentioned there being set up in the middle of 2020. A nice calm relaxing time for everyone involved.
00:03:32 Will Mountford
Walk us through a little bit of who’s involved. What are they working on? And from there we can get into kind of the ongoing research that’s coming.
00:03:39 Prof David Punihaole
Out of it? Yes. So my research group now currently stands at. We have 4 graduate students. My 2 senior graduate students are Maddie and Russell, who are both.
00:03:51 Prof David Punihaole
And local chemistry PhD students, they are just finishing their third year of studies.
00:03:57 Prof David Punihaole
I have a second year student, Lauren, who’s a physical chemistry PhD student and then a my new student at Shakira who is also an analytical chemistry student and she just is finishing up her first year.
00:04:11 Prof David Punihaole
Maddy studies. So I should say Maddy.
00:04:14 Prof David Punihaole
And Russell really built the lab.
00:04:17 Prof David Punihaole
They, and this was very challenging to do during COVID because they did not come in with any knowledge of lasers or optics or anything like that. And it was very difficult to train students with social distancing requirements during.
00:04:32 Prof David Punihaole
COVID, but they are very natural at working in the lab. They’re great scientists, and in the case of Maddie, she is working on trying to develop tools using a technique that our group specializes in called Raman spectroscopy. She’s trying to develop Raman spectroscopic tools to study the structure.
00:04:52 Prof David Punihaole
Of protein aggregates associated with Alzheimer’s disease.
00:04:56 Prof David Punihaole
Russell is using trying to develop similar Raman based tools to study the structural dynamics of.
00:05:05 Prof David Punihaole
Polymers and other types of nanoparticle systems that can be used to deliver genes and drugs perception to cells Loren and secure our working together on a project we’re trying to develop imaging probes that can be used to not only visualize where the molecules are, but actually visualize or or sense.
00:05:25 Prof David Punihaole
Local chemical environments that the molecules are experiencing.
00:05:30 Will Mountford
I can imagine it’s been a busy couple of years for Bioscience generally, but specifically looking at chemical engineered molecules, sensing cellular environments, do you say that this has been something that you’ve seen from the kind of hands on perspective or has it all just been happening and I’ve not noticed?
00:05:48 Prof David Punihaole
No, I I would say you’re.
00:05:50 Prof David Punihaole
Right, at least certainly in this field that I’m in with Raman imaging, so.
00:05:55 Prof David Punihaole
You know, historically people have been using fluorescent probes to tag proteins, for example, and these these fluorescent molecules will emit light that.
00:06:06 Prof David Punihaole
We can then.
00:06:07 Prof David Punihaole
Use to visualize where molecules are located in a cell. For example for microscope, so that works. Been going on for several decades now.
00:06:15 Prof David Punihaole
I think what’s new is the realization that, you know, we can do more than just use a microscope to visualize where things are located. We can actually use these things to visualize the interactions and the structural dynamics of how those molecules.
00:06:31 Prof David Punihaole
Actually, are carrying out their biological functions, and so there’s been work now in the fluorescence field.
00:06:37 Prof David Punihaole
To do this.
00:06:38 Prof David Punihaole
But also since for the United say, the past decade, there’s been increasing recognition in my field that it’s a good thing to develop probes that that can be used in conjunction with Raman microscopy.
00:06:51 Prof David Punihaole
Sense chemical dynamics as well. And so I would really say since you know it seems like since 2020, several research groups have kind of all converged on that, that focus in my field.
00:07:06 Will Mountford
All of that wiggling and jiggling that.
00:07:09 Will Mountford
Well, I’ve got to say, the Punihaole website is a very useful resource for getting me up to speed with all of the content that’s on there and all of the breakdowns. It’s got lots of summaries. So if anyone listening at home wants to have a quick dive into the research that you’re doing, it’s a very good place to start for some of the more in depth details.
00:07:29 Will Mountford
We can crack on to talking about some other resolutions and scales of what you’re working with here. The magnification and microscopy involved there. Is there any useful metaphors or comparisons that anyone listening at home might be able to think? Ohh that comparing the Empire State Building to a grain of rice, that kind of thing.
00:07:47 Prof David Punihaole
So a cell, a biological cell is somewhere on the order of a few 10s of micrometers. You know, that’s about as as thick as a human hair, for example. Molecules are, you know, over an order of magnitude smaller than that. We’re talking about length.
00:08:07 Prof David Punihaole
Scales that are in the order of 10 to the -910 to the -10 meters.
00:08:15 Prof David Punihaole
Very small scales, spatial scales that we’re talking about. These are the kinds of interactions you’re trying to the kinds of length scales that we’re trying to tease, structural dynamics and interactions out of if you take a conventional microscope, say in a high school classroom and you look at some biological cells you’re using.
00:08:35 Prof David Punihaole
Visible light to be able to see those cells right. And so the microscope has a resolving power, given the wavelengths of light that you’re using to see those cells. The microscope has a ability to resolve objects on the order of about 200 nanometers.
00:08:51 Prof David Punihaole
Right. So molecules are even smaller than that, you know, still a factor of 100 / 100 smaller than that.
00:09:00 Will Mountford
Those molecules that turn into human conditions, into human cells, into human disease as well, you know, to kind of ricochet back and forth between the very tiny and the very relatable human scale of things.
00:09:14 Will Mountford
Could we talk a bit about how some of the research happening at your lab relates to, you know, degeneration, Alzheimer’s, how we are connecting the chemical and physics research to the human outcomes?
00:09:28 Prof David Punihaole
Yes. So one of the research interests my group has is trying to understand the molecular basis for a host of different types of neurodegenerative disorders that seem to be linked to protein aggregation.
00:09:43 Prof David Punihaole
The classic example of this is Alzheimer’s disease, where there is a protein fragment that’s called EBITA.
00:09:52 Prof David Punihaole
And this fragment will aggregate abnormally and form these large filamentous aggregates that are known as amyloid fiber.
00:10:03 Prof David Punihaole
These amyloid fibrils will then compact together to form plaques and cease plaques that if you if you do an autopsy on an Alzheimer patients brain, you’ll find that they’re riddled with these protein plaques and it’s.
00:10:16 Prof David Punihaole
Thought that there’s a link.
00:10:18 Prof David Punihaole
Somehow between this abnormal aggregation.
00:10:22 Prof David Punihaole
Of this, a beta peptide and Alzheimer’s. What that link is we don’t know, but there’s certainly some compelling circumstantial evidence suggesting that it’s there.
00:10:33 Prof David Punihaole
So for example, you can do autopsies on on different Alzheimer patients brains and these different Alzheimer’s patients may have different clinical subtypes of the disease.
00:10:45 Prof David Punihaole
And what you find then? If you look at the molecular structure of the fibro aggregates, you find that there’s different.
00:10:53 Prof David Punihaole
Structural types of these aggregates in the brain.
00:10:57 Prof David Punihaole
And so that is one of those types of circumstantial lines of evidence that suggests there’s a link between the structures of these different aggregates and potentially clinical phenotypes.
00:11:09 Prof David Punihaole
It’s also known that these arrogates can cause oxidative stress in cells, and that’s one of the mechanisms by which these aggregates may end up killing neurons in the brain, and that then in turn would lead to neurodegeneration. So our group.
00:11:28 Prof David Punihaole
Studying these arrogates is is very difficult.
00:11:31 Prof David Punihaole
And it historically it’s been very difficult to do. There’s only a few techniques available to study those those aggregates, but all of them are done. All those techniques require that you study the structures of these arrogates outside of their biological environment. So outside of the native brain tissue environment.
00:11:51 Prof David Punihaole
That they’re found.
00:11:52 Prof David Punihaole
And this can have a variety of problems associated with it. Notably, it’s unclear whether the structures that scientists are studying in test tube environments actually are the same.
00:12:06 Prof David Punihaole
As those in the brain.
00:12:08 Prof David Punihaole
It’s also unclear that if you try to purify these aggregates out from the brain to study them, that is it’s it’s unclear if that purification process can actually perturb.
00:12:19 Prof David Punihaole
The structures and So what you’re studying in the test tube environment is not necessarily what you’re studying. What’s clinically relevant in in brain tissue?
00:12:29 Prof David Punihaole
So that you know, was a big motivation for my group where we’re trying to develop tools that are structurally sensitive to that can be used to study protein structure and we’d like to develop those tools so that there is non as there as non invasive as possible where we can actually use them to directly.
00:12:49 Prof David Punihaole
Study these proteins and their aggregates in cells in brain tissue and things like that.
00:12:58 Prof David Punihaole
So that’s that’s the ultimate motivation for our work, because our thought is if if we can understand the structures of these different aggregates.
00:13:07 Prof David Punihaole
We could potentially link the structures of these arrogates to different clinical phenotypes you observe in these diseases. We can understand really the structural basis for the different toxic activities of these aggregates in the brain.
00:13:23 Will Mountford
And is that something that you see as being a fruitful avenue of research? Are you just getting started?
00:13:27 Will Mountford
Is there anything that’s come out of that so far that appear, or another professional might be able to read more about?
00:13:37 Prof David Punihaole
So the big picture idea that I I described, we are just getting started. I do think it’s a fruitful Ave. because I, you know again the field the amyloid field has been stuck on trying to prove that there’s a direct link between the structure of these different types of aggregates and toxicity.
00:13:57 Prof David Punihaole
And this field is very… there’s been debates raging in this field for the past two decades.
00:14:03 Prof David Punihaole
It’s, you know, it’s some. Some groups argue that it’s not these these large plaques that are causing issues. It’s actually smaller precursor aggregates that apparently interact with cells and and causes cell death. And so, you know, you, you for for almost 2 decades. It seems like there’s been multiple.
00:14:24 Prof David Punihaole
Different camps arguing about about these things and all of these questions I think could be solved if we had tools to just be able to directly study these aggregates in their native biological environment. It’s, you know, it’s high risk, but I think it’s worth the pay off if it’s going to finally resolve some of the big questions in the field.
00:14:43 Prof David Punihaole
We have been working towards the first steps towards addressing these big questions, right. So, you know, recently my group developed a technique where we can use Raman spectroscopy to measure select structural parameters in these fibro aggregates, things like different bond angles.
00:15:04 Prof David Punihaole
For example and we can then actually use these structural parameters that we measure.
00:15:10 Prof David Punihaole
As constraints in computational simulations, we actually then constrain
00:15:16 Prof David Punihaole
The experimental parameters we measure.
00:15:18 Prof David Punihaole
We we feed that into a simulation where we constrain the structure of our protein aggregate. We run that simulation and we get a set of structures that resemble our experimental data.
00:15:31 Prof David Punihaole
And that’s how we actually visualize the structures of these different aggregates. And we’re hoping we can at least translate some of that methodology in the near future towards imaging applications.
00:15:45 Will Mountford
Now we’ve mentioned it a couple of times, but I think it’s, you know, worth going over some of the the concepts and the experimental setups behind the technology here of what is Raman spectroscopy and what that data could hold in the future.
00:16:01 Prof David Punihaole
So Raman spectroscopy is a very old technique actually. I think it now it dates back to the 1920s.
00:16:09 Prof David Punihaole
It was discovered by Sir CV Rahman, who was an Indian scientist, and it’s a light scattering meta.
00:16:17 Prof David Punihaole
The idea is you use laser light typically to excite scattering in molecules.
00:16:26 Prof David Punihaole
Now most molecules are going to scatter the laser light.
00:16:31 Prof David Punihaole
So there’s no change in energy between the incident laser light you use and the scattered light that you see. So if you have a green laser, you’re going to see green scattered light get emitted from the sample.
00:16:44 Prof David Punihaole
This type of scattering, where there’s no change in energy between the incident and the scattered light is called really scattered, and it’s actually the reason why the sky is blue.
00:16:54 Prof David Punihaole
Because there’s a wavelength dependencies like.
00:16:58 Prof David Punihaole
The more blue the light is, the more efficient the scattering process. So the reason why the sky is blue is sunlight comes in to the ATM.
00:17:06 Prof David Punihaole
Where you have a nitrogen molecules in the atmosphere and they can scatter the blue light much more efficiently than they can scatter the red light, and So what we see on Earth on a sunny day is is blue light. Due to that process. Now a little bit of that light. You know, maybe one in a million photons.
00:17:26 Prof David Punihaole
Is going to get shifted in energy and the light is scattered and so what’s happening there is you’re you’re incident. Photons interact with your molecules and they drive vibrational motions in the molecules.
00:17:41 Prof David Punihaole
And so as a result, energy is being used from the light to excite motions. Those wiggling and jiggling of those atoms that we talked about earlier as a result of that exchange of energy, the emitted scattered light is going to be shifted in energy.
00:17:57 Prof David Punihaole
And so that’s going to manifest as a change in the frequency of the light that we measure.
00:18:02 Prof David Punihaole
So this is the Raman scattering process. And because our technique probes molecular vibrations, we can actually.
00:18:11 Prof David Punihaole
Use those vibrations to learn about how the molecules are interacting with their local chemical environment. Because molecules will change their structure as a result of these interactions, and that will cause them to scatter the light differently. So this technique is very old. As I said, it dates back to the late 1920.
00:18:32 Prof David Punihaole
Needs, but it’s historically it was very challenging to do and that was because the technology really wasn’t there to make this an easy type of experiment to do.
00:18:44 Prof David Punihaole
So it really wasn’t until the development of the laser that Raman spectroscopy became more popular and that so it was discovered in the 1920s. But the laser was invented in the in the early 1960s, so it took several decades before lasers were invented. And the reason why lasers were so important was because they can.
00:19:04 Prof David Punihaole
Emit much stronger light that can be used to excite Raman scattering, because it’s such a weak process and so.
00:19:16 Prof David Punihaole
As a technique to study biological molecules.
00:19:19 Prof David Punihaole
And you can use a laser to excite the vibrations of molecules like proteins or polymers, and then you can collect that scattered light and send that scattered light into a device called a spectrometer. And what the spectrometer does is it separates that light into its different color components, and you can then.
00:19:40 Prof David Punihaole
Image that onto. And that’s how you measure what we call a lament spectrum. And that technique works.
00:19:48 Prof David Punihaole
It’s still a very slow and painful process sometimes to do some of these experiments, one of the problems with conventional Raman methods is that a lot of biological systems, including cells, naturally.
00:20:04 Prof David Punihaole
So they emit fluorescent photons and that fluorescence is rather strong compared to the Raman scattered light, and so you often will detect both the the natural fluorescence of the cell as well as the the scattered light from the Raman photons, and that can complicate and.
00:20:24 Prof David Punihaole
Overwhelm your Raman signals. Just it makes it very challenging to do and it’s because of issues like that that you can’t do live cell imaging with traditional Raman methods. And that’s why for a long time, biologists really did not want to use this.
00:20:41 Prof David Punihaole
However, around 2010 the field started to change. And that was because the technology and detection systems had been developed so that you could do something called stimulated problem scattering.
00:20:55 Prof David Punihaole
So the idea here is you don’t just use one laser to excite the scattering, you use two laser beams.
00:21:02 Prof David Punihaole
And you tune the wavelengths of these two laser beams to particular vibrational resonances of the molecule, and you can then stimulate the emission of the scattering process.
00:21:15 Prof David Punihaole
This has made it now feasible to do live cell real time video rate imaging of biological cells, and so that’s now something that my group is moving into to to further develop this, this technique you’ve actually been constructing A simulated Raman microscope and.
00:21:35 Prof David Punihaole
We hope in the next year or so that we can get our first live cell data from.
00:21:41 Will Mountford
Well, another Ave. of research that the Punihaole website spells out quite neatly is the drug delivery Avenue of research stuff. The quinine coupling as an example of your work that led out to me. So wondering if we could kind of hop from the post mortem cells to the live imaging of cells and.
00:22:00 Will Mountford
How the Raman spectroscopy and the amount of data you can get out of the scans that happened there are informing drug development and.
00:22:08 Prof David Punihaole
Yeah, so this work really stems from my time as a postdoctoral researcher at the University of Minnesota and Minnesota. There was a great cohort of researchers who were trying to develop alternative.
00:22:21 Prof David Punihaole
Methods to deliver drugs and therapeutic nucleic acids to cells for cancer, gene therapy type applications. So currently the gold standard method is to use viruses to deliver genetic cargo to cells to do gene therapy.
00:22:41 Prof David Punihaole
Viruses are great because they’ve essentially evolved to do that task.
00:22:46 Prof David Punihaole
But as you and and the viewers will know, like viruses have a major drawback in the fact that they can elicit immune responses in patients.
00:22:57 Prof David Punihaole
And actually, there’s been a number of high profile cases in gene therapy trials where patients have actually died as a result of using viral vectors for gene therapy. Patients have also suffered other complications. They’ve developed carcinomas, sarcomas.
00:23:15 Prof David Punihaole
And other health issues. So viruses as good as they are at at efficiently delivering their their cargo to cells to enable efficient gene therapy still viruses, they are still viruses. And so the health risks are something you have to seriously weigh as a clinician.
00:23:37 Prof David Punihaole
When when you’re conducting one of these therapy trials?
00:23:42 Prof David Punihaole
So, you know, researchers at Minnesota and and all across the world have been trying to develop alternative, safer vectors for delivering.
00:23:53 Prof David Punihaole
The genes to cells and polymers are one such delivery. So many people I think are probably familiar with polymers. It’s, you know, polymers can comprise the materials that coat your non-stick pan rubber from a tire, that’s a form of.
00:24:12 Prof David Punihaole
Polymer they are basically just long chains of molecules that are linked together to form a larger system that we call a macromolecule.
00:24:22 Prof David Punihaole
But you can have polymers that are safe to use on cells that are biodegradable, and these types of polymers seem to be really useful for gene therapy.
00:24:32 Prof David Punihaole
Drug delivery application. The problem, despite their potential advantages for being safer.
00:24:41 Prof David Punihaole
Is that they are nowhere near as efficient as viruses at delivering their cargo.
00:24:47 Prof David Punihaole
And a big reason why is because we don’t understand how they actually work.
00:24:51 Prof David Punihaole
So there’s a lot of research groups trying to understand exactly how do these polymers interact with cells. How do they even get to the cell so that they can get internalized and then deliver their cargo to the appropriate compartment In the cell.
00:25:07 Prof David Punihaole
There’s a lot of unknowns there and part of what makes it so hard to study the mechanisms by which these polymers carry out their gene and drug delivery functions is you just don’t have good tools to study these polymers.
00:25:22 Prof David Punihaole
So I saw an opportunity here to try to make a difference with with Raman spectroscopy, the system that we initially worked with was I had a collaborator, Teresa Reinecke, and her research group had developed.
00:25:36 Prof David Punihaole
These polymers that contain quinine, quinine is a molecule that is found in tonic water. For example, it’s it’s the oldest drug known in the western world that was used to treat malaria. So it’s it’s very biocompatible and it turned out that incorporating quinine into their polymers really enhanced.
00:25:56 Prof David Punihaole
The gene delivery efficiencies of these polymers.
00:26:00 Prof David Punihaole
And the big question was why? So we weren’t sure we could actually answer this question, but we felt that we could use our tools to understand a little bit of how these polymers worked and the the first question we asked was how do these polymers do you know where do these polymers deliver their cargo in cells?
00:26:20 Prof David Punihaole
Can we use our Raman spectroscopic tools to actually track the cargo release in the.
00:26:26 Prof David Punihaole
cells, so it turned out the Raman signals that we found for quinine were very sensitive to probing how quinine interacts with DNA. And we could see characteristic shifts in the scattering frequencies of these quinine signals to actually.
00:26:47 Prof David Punihaole
To figure out the different ways that quinine could essentially interact with the DNA they could, there’s a different types of what are called non covalent interactions that molecules can engage in and we could actually figure that out with with quinine.
00:27:01 Prof David Punihaole
So we then use this signal to actually monitor in cells where the cargo was being released, and so we published a paper in 2020 on this Teresa Ranicki’s group where we showed that we could actually visualize these non covalent interactions directly in the cell for the first time.
00:27:21 Prof David Punihaole
Using an optical microscope, so this is a really powerful advancement.
00:27:25 Prof David Punihaole
For the microscopy field.
00:27:28 Will Mountford
I mean, from everything that we’ve talked about, it really seems like your research sits at the junction not just between biology and chemistry and physics, but also the technology aspect. I mean mentioning that it takes 40 years for someone to invent a laser, so that Raman spectroscopy can be done, and then another 50 years for someone to come up with stimulated spectroscopy.
00:27:48 Will Mountford
How do you see the kind of wave of, you know, technological advancements moving at whatever pace they are moving at and all of the excitement about AI?
00:27:57 Will Mountford
Way in synthetic biology, whatever you know, the next big thing in technology is how do you see that coming to your work and what kind of doors does it open?
00:28:07 Prof David Punihaole
Yes, this is, I think, a very exciting time right now to be in this field
00:28:11 Prof David Punihaole
It is very rapidly advancing, you know, I should say that you know my group, we focus more on applications.
00:28:18 Prof David Punihaole
Where we’re using the methodology to study particular biological problems, but there are have been other groups, most notably.
00:28:27 Prof David Punihaole
Jishan chens group at Boston University Layman’s group at Columbia University, for example, who have been really pioneering the technology development to make it easier for researchers like myself to do stimulated Raman engine. So yes, I think you know from a hardware point of view, getting lasers that are much more reliable.
00:28:49 Prof David Punihaole
And less finicky.
00:28:51 Prof David Punihaole
Has been really important and so those lasers are now available commercially to researchers, getting better detection systems so that you can measure these weak Raman scattering signals has been absolutely crucial, in particular for simulated Raman, because.
00:29:08 Prof David Punihaole
It’s not as straightforward to detect the scattered light as it would be with a conventional Raman system. You have to do a lot of electrical engineering tricks that you have to do.
00:29:18 Prof David Punihaole
Which a chemist would not necessarily know how to do, but you know, fortunately the folks I just mentioned have figured this sort of stuff out. And so that technology is now becoming commercially available as well.
00:29:32 Prof David Punihaole
I anticipate within the next 10 years, there’s probably going to be commercial stimulated Raman setups that will be made available that will be user-friendly enough that biologists could probably use. And then, you know, you mentioned the software side of things. Once you get all this data, particularly if you’re doing live.
00:29:52 Prof David Punihaole
Cell imaging work. You’re going to be collecting potentially terabytes worth of data.
00:29:58 Prof David Punihaole
How do you make sense of that? All you can’t individually hand process.
00:30:04 Prof David Punihaole
All that data and so I think that’s where automated approaches are going to become useful. There’s been a lot of work from other researchers who are using machine learning techniques to denoise the the data that’s collected to be able to help make sense of the data. So the the data can often be very complicated.
00:30:25 Prof David Punihaole
Difficult to interpret and so there are definitely algorithms that people are developing to try to help interpret the data.
00:30:33 Prof David Punihaole
In terms of my research group as chemists and not as engineers, we will.
00:30:38 Prof David Punihaole
Certainly be using these.
00:30:40 Prof David Punihaole
These technologies, when they’re made more accessible to help us answer questions in biology and chemistry.
00:30:54 Prof David Punihaole
You know, to recap, my research group is interested in helping to change the paradigm by which we do biological imaging. Fundamentally, it’s not enough for us to know where a protein, for example, is in a cell at a particular point in time. We really want to understand what that protein is doing in the cell.
00:31:14 Prof David Punihaole
We need to understand the chemistry, so we need to be able to understand how that protein is interacting with its local environment or carrying out or what kinds of structural dynamics it’s undergoing to carry out its.
00:31:27 Prof David Punihaole
And we believe that our our Raman spectroscopy based tools are amenable to to getting that kind of information.
00:31:36 Prof David Punihaole
So in particular, we’re interested in developing these tools that we can answer questions related to understanding the molecular basis of neurodegeneration, particularly for diseases like Alzheimer’s.
00:31:48 Prof David Punihaole
We’re also interested in understanding fundamental molecular mechanisms behind which polymer and other types of nanoparticles deliver drugs and therapy and plic acids to cells for cancer therapy applications. Then, finally, we’re interested in just understanding fundamental aspects about biology.
00:32:09 Prof David Punihaole
Most notably, how do you structural dynamics of proteins?
00:32:13 Prof David Punihaole
Differ and how are they regulated in the cell compared to test tube based experiments which is what most people tend to do?
00:32:21 Will Mountford
For anyone who’s listening to this episode, is there anything that you think they should do next after listening?
00:32:27 Prof David Punihaole
When people think about science, they always want to know what the payoff is. And sometimes with fundamental research, it’s very hard to see what the payoff is.
00:32:36 Prof David Punihaole
Right. So we’re trying to develop a new technique and it’s not quite clear that that technique always has an application. Clinical settings, for example or.
00:32:46 Prof David Punihaole
Something like that. But I think it’s important for people to realize that.
00:32:51 Prof David Punihaole
Developing new technologies in science is vital towards advancing science and making breakthroughs. The microscope and the telescope, I would argue, were two of the most important developments in science because it allowed us to study things that we could never have seen before. Things on the very small scale.
00:33:11 Prof David Punihaole
Like cells and things on the very large scale like planets and Galaxy.
00:33:17 Prof David Punihaole
Right. And had those technologies not been developed, we would not know about a lot of our universe, right. And so in that same vein, there are researchers like myself who are trying to develop new imaging tools that can allow us to probe new areas of science.
00:33:34 Prof David Punihaole
So people can go to my website to find out more information about my research go to The Punihaole Group.squarespace.com.
00:33:44 Will Mountford
Doctor Punihaole , thank you so much for your time and hope to speak with you again sometime soon.