 The title of my presentation is the Opportunities for Syncrotron Imaging, Internationally and Locally, and I had a hard time figuring out what to put in this presentation because there's so much you can do. And I mean, we have been talking about functional and anatomical imaging and we've been talking about, well, in the imaging, X-ray imaging and really depending on what you want to do and what you want to see, you can use certain methods. And the syncrotron, well, it's just a source of X-rays and you can do anything, depending on which be mine, you go to. So I tried to put together some slides to give you some idea of some examples. The next presentation after me is Goran, we'll be talking more about imaging and hopefully also a bit of in vivo imaging, but yeah, I'll try to cover some topics of what is possible. So first of all, the facilities, I mean, we have the, assuming from year and year by max four, and there is some possibilities there for imaging, not so much for sure no in vivo imaging yet at least. And then for micro CT, there are some beam lines that are coming online now so in the next year or so, we will have more and what you need is to do microtomography here. At the same time, you can also use other facilities such as the ESRF in Hannibal, Pixel 3 that's in Hamburg, Diamond that's near Oxford in the UK, so they use in Paris. We found also the SLS, here's the Swiss life source where Goran is traveling in from, and here's our own neighbor here, max four. As I said, a single turn is just a really a source of x-rays so we have a big storage room here at four and one possibility to do in vivo imaging would be a medical imaging beam line. So that's the proposal that we have in the roadmap of the max four development, but it's not a beam line that exists yet. In the meantime, there are other beam lines that you can use, there's the Denimax beam line here, where you can do imaging, there's the Nanomax beam line here where you can do micro and nano probe imaging. And then there are other beam lines for approaching crystallography and spectroscopy so you can do all sorts of things and any of those could be related to drug development and discovery. So, well the title is imaging of this workshop so I will focus on imaging, but just to say that that's spectroscopy and crystallography of course are all the tools that are available. So what's what's the difference between using Sumitron radiation rather than normal x-ray tubes and the other imaging methods that Neil just introduced us to. Well, it's my crash course in physics one slide on what's what's happening with the x-rays. So basically, when you have the x-rays coming in, well we can have absorption and we can have scattering in the material. So that basically gives us some attenuation of the x-ray beam and it gives us a normal x-ray image that we see in the clinics for using a normal clinical CT scanner or micro CT scanner. So this happens as well. So we can get a phase shift of the incoming x-rays and we can get fluorescence also from this. So, so there are other possibilities of doing imaging. And basically when you have a well behaved source of x-ray radiation like you do at the Sumitron, then you have many more possibilities of getting nice images out. So just to say that that's actually some some physics behind going to new imaging with others. So one one opportunity is really just to do micro CT but just better higher resolution better contrast. So the contrast would come from using the wave properties of the x-ray wave and then the high resolution is just because you have much more photon flux and higher resolution So here's a recent study from Nature Methods that we have in the last year where they have been imaging different types of tissue, long brain, kidney, spleen, heart, and some really really high resolution so they have been imaging the entire organ but then the very high resolution so you can really zoom in and get all the information you want. And of course this is not an in vivo method. This would deliver very high dose rates. And the data sets here also really big because again if you have very high resolution of a very big volume then of course the data is just large. So, so it's a very powerful demonstration of what you can do with Sumitron imaging. In terms of micro CT. And yeah, so I mentioned the physics a few times and I'm not talking about this just to say that we can use the x-rays not only as these beams of particles but actually as new kinds of waves, you can get a refraction and use this to get better contrast. So what, what kind of research can we do what type of data do we get out. So one thing is that we can do a virtual sectioning so we don't need to physically slice our samples. So instead of, well, comparing to the normal strategy, whether we need to physically slide in thin slices to cutting, then we can image the entire volume and get this virtual section in any direction you want. So what could this be useful, well this can be useful for obtaining a much better understanding of three dimensional structures and this is an example from cutting. It's a sexy foundation in the long run, so I think it's from our attention. Getting the 3d information to high resolution is really something where we're just just doing normal micro CT but one step further so we can do even better resolution even better contrast so as you said Leo it's just the next step that we can do this. And then of course when we have all of this information we can also do data extraction so we can do statistics and this example here in the corner is the sun axons. We can measure the diameter and the shape of these axons for individual actions and then we can extract all this data from the entire volume so so we get lots of data out that we can do statistics on. So, better statistics and better volume metric understanding of microstructure structures. These are the main advantages of doing singleton based microscopy. So when I say microscopy is kind of opposed to to the to the envy imaging. I don't know if there's a clear distinction between microscopy it has to be extremely evil. And but of course it's it's it's why we, we, we would normally use other types of my first piece in the visual light relation microscopy or. The other message for for for looking at structures that are well in the in the micrometer range. So, so this is a message needed to do to do this in three dimensions. So I, I look a bit at the workflow, I mean going from from in the imaging and into using what would use the singleton radiation. We can use the, the singleton for in legal micro CT, and then maybe the resolution limit there would be a micron or a few microns because then we get into the problem of too high radiation dose. So if you want to go to higher resolution and do microscopy, but then we have to usually do it ex people and then we are in this virtual here of normally when we do microscopy. This is what we do what we expect some tissue and we dehydrate and embed in some medium. And then we slice it, but just before the slicing, we could do this extra micromography and get the volumetric information. And then we can continue with our work that will do the slicing and do this. And we can even do the rest of the process. So that's the method that I didn't talk so much about yet, but that's good. I will come back to the fluorescence based engine. Here's an example here. I can write it black. So an ongoing study. Here is to show you the difference between micro CT with and without the face contrast just using the high coherence that's an extra beam. We can do face extraction so this would be the image to the left is without the face retrieval. So basically the only looking at the normal attenuation and then you see the difference in the contrast again can be at the singleton. So we can get really a lot of contrast in the soft tissue. Which is, of course, a big advantage when we, I mean the image would solve before from the, from the, looking at the bones in the micro CT with that's something you can do because of bones that could contrast to the soft tissue. But if we wanted to see features in the soft tissue, then we need to have this space contrast. And while hearing the example of corroded like what we have been doing MRI images of the extractive class so this is human class that have been extracted. And we can see the resolution is somewhat limited. So we can go to the to do micro CT, we can image again the entire flat. So this is the diameter of five centimeters or something like this. And then we can zoom into regions that we can interchange so maybe the contrast here is not so much in particular with the sun that coming in. So we can really zoom in and get cellular resolution in the intact sample. So, so without physically slicing it. Well, it's currently black. Yes, it's from the archery from a patient. So when you have like here, then patients are operated to to remove the risk of the road trip to black and and the black that has been taking out of patients these are stored in the bio bank. And we can study them without any ethical problems with with the patients and it's actually these samples here are from for living patients but have been removed. Yeah, so here's a bit better resolution so so you can see here in this study we have the entire fact that has been removed from the patient. So here's the length here. Well, three centimeters more. And then we can do high resolution study but we use the entire thing and then we can zoom in on small regions and do even higher resolution to study particular regions. So here's a corner where there's a risk of rupture. We can look at the, at the exact methodology here and compare it then to so afterwards we can do the slicing and then we can do the normal staining and so on with the strategy, and then we can do one to one comparison of our symptom data. So I would say this is, this is a direct extension of micro CT but just with better contrast better images higher resolution. And then of course we can, because we have the volume metric information, besides comparing to the 2d slices that we can study. And we can also make make renderings and videos and yes video. So what you see when we're going through this volume gear. This layer here is this elastic membrane. And when you can image this multiple slices, then we can segment it out and do a visualization to, to see the three dimensional morphology of these structures. So this is a membrane as I said, another example of the new vessels. So again, this is the corner here of the canvas. And inside here we can see lots of different structures that's many different, I mean, we cross this and that looks like in this also we give you where they have been new vessels forming, so we can segment out these new vessels and image them and we can then get an idea of where do the business come from and why do they go to something that's completely impossible to see with them. But it's just truly slicing. And this was done without any staining of the sample. And then the last half of my presentation will be on fluorescence imaging. So basically, using this other method that when we're exciting the atoms in the in the sample with our x-rays, well then we can also get actually fluorescence out. There's also a very powerful method and again it's, you can compare it to fluorescence microscopy that you know visible light. Except that here well we're post sensitive to the elements that are already there and we can also mark with specific markers and look for particular elements. So, so here this is from the Australian Symmetra. So it's a tadpole and the other imaging software Symmetra and see in this color coding here, so red, blue and green to trigger the distribution of these elements in the tadpole. But I will show you some more examples from our studies here as well. So the, what you need to be aware about these. H-ray fluorescence is that this is now no longer full field imaging, but it's a scanning probe. So we're focusing now the H-ray beam to a small spot and that's the scanning and then we can get fluorescence out in the very same way that you can also do with an electron microscope. So you have a rest scan of our sample and then we can have fluorescence detection and we get these milios out and at the same time we can also record the transmission so we get also the H-ray transmission images. So here's again for the same samples before the chromatic plaque and then after we have done the sectioning and the histology, then we can identify regions that we want to look at with the high resolution H-ray fluorescence microscopy. And then we can map out some, here's one particular region, the S1 region here and we see the image up here to the right, that's the H-ray transmission. So we see this something that will show up in the microstatic image as a healthification. So these are some absorbing structures and they are sure enough mostly calcium but also a lot of iron you can see. And we can see the distribution of zinc and sulfur and other elements that are there. Without adding any steaming, so this is images up here they say but this is a neighboring slide. And they're quite specific, that's a high sensitivity and high physical sensitivity. So this is measured out in the frames grams for micro use graph. I don't know if it corresponds to a medium or a micromobile but it's something that can be figured out because it's a quantum method. So this is the resolution here. Now when we're doing the scanning probe and it's extremely then we can also go to very high resolution so we can do a titographic imaging so this is a several years ago now, what have been doing high resolution imaging of the single cell and then inside the single cell mapping out the distribution of the phosphor, calcium, calcium and sulfur. So, again, it's, I don't know what you would call functional imaging but it's this distribution of the elements that are involved in the functions in the cell. So, and then you can, you can do this both in truly and in three dimensions. I just threw in this slide as well during the, the morning session, because there wasn't talk about staining so this sample has to stay with us. And then also staining with the catadydinium and look at why does the catadydinium end up if we have turned some MRI based and then we can see if the catadydinium city inside the cells outside the cells of where they sitting so so this is for us in city because the sample was made for electron microscopy. So again, it's like the nerves that I showed you the one of the pre-disposed that have been cut and then these thin sections have been the image at the nanomax line with the fluorescence. And again, we can look at the other elements as well. But as I said extend this into two three dimensions so we can do scanning for imaging of a specimen that has not been cut, rather than being punched in some smaller size. So samples that are millimeter in diameter on this range, you can actually do mapping in three dimensions of the elements and see the distribution. And yeah, it's really that silicon, phosphorus, chlorine, calcium, magnesium, all the different elements that you can come out. And yeah, I think this is what I chose to include for now but I mean as I said you can do anything where you could do spectroscopy you can also when you have a scanning probe so you can raster scan and do imaging as well. So there are many of these methods that are not only spectroscopy but also imaging as if you choose to do it like that, and that you can do the solution. Yeah, so basically Michelle, Jason and 3D and some of the key points that you can really benefit from. Thanks. We have two excellent questions from Char from online audience. Iran, a PhD student from medical faculty asks, I was wondering about all metric analysis, if it is really accurate since the processing also causes tissue shrinkage, is there a method to normalize for that shrinkage? Yeah, the shrinkage comes from the fixation, I mean, so it's one obvious answer that is to do the imaging before for the fixation which is also possible. It's possible to get on frozen tissue. Yes. Yeah. So I mean, in most of these studies we're doing it on the samples that are already fixated and in paraffin because these are the samples that are accessible to us in the Biobank. And so they're very easy to get access to, but it's perfectly possible to do it also on the ratio of process samples. So there is a cross the top during the measurement. I mean if you need to be frozen during the measurement you will need to have a prior setup. There is another patient. I wonder if it is safe to use in return to image patient compared to death or CT. Well, I mean, imaging patients, you cannot do it at this high resolution in any case. So, so you're you're really limited by by the variation dose. So there are, I mean, for instance in the single team in the novel, they have been doing imaging patients. And sure enough, because you have the single team, you can choose a single energy, and you can be more, more dose efficient because when the you get rid of all the low energy set that because variation dose. So, so there are some studies showing this, but but I think most students say that that they're not meant for for patients. So it's a big hassle to get the patients to the to go to the cemetery. Now we have questions. I was just wondering. Well, this is very cool. I was just wondering, like, what is the main advantage of this technique. Against the, or in front of the regular standard. You just show that the black. There was not so much difference at some point you need to distinguish for example, except themselves. I think that the older the methods have their own advantages. They are all unique in the sense that if it's a particular thing you want to see that's probably a method that can show you that. But the point is here that well, I mean, if you're doing slicing, then you don't get the free information for the clock. So, for instance, you cannot do spicy because if you have communications, the communications will be pulled by the knife and destroy the spices so that's a high risk of the great petition. So basically what you do is, is you decalcify the tissue before slicing it and then you can see the world that will cost you. But when you do a non-destructive imaging, without cutting, then then you can see the solution. For example, but what's holding the community back to not use this. I think what is what is holding I mean the major barrier is that it's a little bit difficult to get access when you don't know what it is and what to do and how to do it. I mean, I think that's basically what we did today to lower that barrier so that we can get more people. Because it's really not so difficult. Martin and I have been working with lunch together. And I think when you're working on branching structures like the lung, which is difficult to understand from a 2D section. I mean, you can see the view that you get for the syndrome, which you get really high resolution in a cube that's 3.5. So, I mean, you can follow vessels and you can see details. And then you can section and then you can do inside the head of the patient or things. I'm just curious about your or the thinking at Max for when it comes to your image and being once in some environment. If you want to do for example, in the end without limitations and restrictions. Well, I mean, I think at Max for Well, we have put in a proposal to have this biomedical in really be mine and has come through the, the whole proposal system of Max for and that roadmap, and this be mine is the top of the project. So, so basically what is missing now is this funding to build it, which is a significant structure. But I mean, it's perfectly possible and that's definitely a will to do it. And right next to Max for is the CMU animal house, so that would be a very good possibility to to have this change between animal house and the syndrome. So I think it's something that's perfectly possible. But we're talking about animals. Well, I mean, I think the large animals is probably wrong. Live animals. Yeah. So it's excise tissue. That's what's going with Ryan. Yeah, I mean, if it's excise and it's much easier to handle, which is why that's what we're doing now we can easily bring the samples to our system. I mean, it's just easier, but it's definitely possible. You know, at that max, they don't have a possibility to handle. Yeah, I said, it's possible. So, so they're taking the process now I think by the fan max. There's also been a discussion for them. It was quite clear that can do it. Because that was also being built as a material. But it's quite clear that there's quite a large interest in life. So the direction of the interplanetary process has changed somewhat to be able to influence life sciences. But if you haven't been able to control a bit of it. And this is why it's so important to have the team for life sciences. And actually, the idea is that you have two end stations, two different types of emotions. My studies, but also what issues. And then to complement it also with the other things that we have. Yeah, thank you. Thank you. So I can appreciate you're setting up an experimental condition for the character study takes a long time. But my question is, once it is established, established, what is the disability to use that method to scan through the different samples of the biobank that does exist? I mean, is it feasible to look at many what's the time link for the analysis. Any questions for that question. No, I mean, I'm not working with traffic tax for working with non samples, but usually if we go more into the context, we have 40 hours to be able to scan the 100 parking blocks. Okay. And then we scan maybe three areas. We usually section of the top section to the side of this area. We get a lot of data. I would say this is difficult to feasible. And so right now you're going through the normal proposal around so is every half year we apply to be in time and then you can be happy later. But I mean, also mentioned that you could do this on a faster track. I mean, maybe soon to say coming up with different proposal schemes and you can get easier access to. One of the scans that we do takes only two minutes, three minutes, with the rest of it at the moment. With a micro seat, you could get close to that. We'll take overnight. That's amazing. We have more time in the kind of discussion. Thank you for asking questions for the panel discussion. Thank you so much for your talk.