 Some of you may wonder in sitting in the audience, I mean, this is a workshop dedicated to synchrotron imaging at the end. So why are we talking about all these other imaging modalities all the time? I'm sure there's at least one thinking that. And the reason for that is to, I mean, at the end, we want med max at max four. And we have to find arguments for that. And one argument, I guess, is to engage with different pharmaceutical companies to promote the funding of med max. And to do that, we have to offer them something they don't really, they don't already have. So that is kind of why we're having this stop of the workshop and the working group to make a common knowledge of what they actually already have and which they don't want to pay extra money for. And you see, I pressed the button. So we're making progress in many respects here. So this graph again, as Leif pointed out, there is many imaging activities going on in the pharmaceutical companies. They have their own equipment. They can set up their studies as they do. They have a very skilled personnel to do the imaging. So on the shrink control, we need to do something quite different. And in order to know what is quite different, it's my duty to learn or to teach. You can say what we do with the normal, normal or if you can call it the standard model imaging modalities. And the reason why we have imaging at all in the pharmaceutical industry is because they are adding some value which other method doesn't do, like astrology, for example. So one very important point is that it's in vivo. And I know if we talk about synchrofront imaging, obviously there's lots to do ex vivo. And once it comes to in vivo, everything gets a little bit complicated. But that is what this working group is kind of aiming for, the complicated in vivo phase. Because you don't have to kill the animal which can make these studies to be longitudinal. And also the imaging provide this three dimensional data. So you get a lot of data at the same time. And if you want a good imaging modality, it should both be able to assess and classify disease on animals and then on patients as well. If it's good and also the treatment response so you can see the effect of new drugs. Try new drugs, see the effect. And the translational aspects mouse to man. So I'm starting with the three different modalities to kind of show this is what we can do with normal imaging equipment. So this is just the micro CT. And the micro CT provides contrast like X-rays, but tomographics to get the three dimensional volume. In this case, we see just a one slice out of that. And obviously there is high contrast in the bone and there is high contrast in the lungs. And then there's no so much difference between the different soft tissues in the CT image. So bone is something which looks very great and have the high contrast. So that can be explored with micro CT. In this particular study, they have a spatial resolution of 18 micrometer, which is quite good. So if you wanna do something better, it has to be certainly higher spatial resolution than this. And what they do is to measure the force of the mouse and then they extract the bone. And by that we can measure the volume of both the soft tissue and the bone in this region. Why would you do such a thing? Because there are one disease called rheumatitis where there is growing bone and it's a very common disease. And maybe this is the thing is you want an animal model to replicate what's happening in you. And that they have. So this is the scar and then different weeks after they introduce the disease because this is kind of typically, you have when you buy animals, they are healthy. That's the point of course. And then you have a disease model or a disease mechanism that you introduce. So first you have to make them sick in order to treat. Then you can see that it's going worse in the bone. And it is all measured by procedure. So what is the study then? Well, you measure the volume of the bone and you can see it's what a naive mice, nothing happens. They're just healthy. In the sick group, you get the bone volume goes down in the base of this disease. And then when you start the treatment, you see that there's a difference between the treated group and the un-treated group. And if this was a study in the pharmaceutical company, there would be a fork, which is the new drug which you compare them with, is that better than traditional, which is steric? If you look at the literature, you can of course never find these studies where the fork is unknown for the drug is because that is seldom published. It's a few one, but then it's long after the drug is being explored. So that's how you do. And this is quite efficient with a simple micro CP which you can have in-house, which is of course a lot cheaper than the synchroforms. So with this example, I would argue that you can do all these things from, it's in vivo, you assess the disease, the treatment response, and it's obviously longitudinal. So that's the micro CP. What's the limitation then? What can we do a lot better? Obviously there is the spatial resolution, which in the micro meter range, that can be improved a lot with synchroform. Limitation as such is that from the CT, it's only anatomical data and not functional. And then sometimes the radiation dose is a problem. In this particular study, it was one grain, which is a lot, but it was only to the feet. So it doesn't really matter that much. But obviously, if you irradiate the whole animal, like in the stomachs of the lungs, you have to keep it the dose of less than one grain, till the 50 for mice is about 4, 6, 5, so to give you a point. Then we have nuclear medicine. So nuclear medicine is something we do with pet cameras and spec cameras and the idea is that you have a radio-nuclide, could be many different ones, and that gives you the signal, the thing you measure. And then you have the probe, which you attach on that radio-nuclide and the probe decides where the radio-nuclide go. And then you get these images, uptake of the specific probe, what that is going. And very common applications are, of course, the glucose of 18-4 chlorine that goes to, actually, anything that consumes energy, which tumors do a lot, they like the sugar, so you can see that. But it can be a lot more specific. So a probe like the analyte that goes into your brain can be used in studies of Alzheimer's disease, or a chol-A1, which, Karin, did you, did you show that? No, I didn't. No, you didn't, okay. That goes to chol-A1, which can be interesting in fibrosis study, or PSMA, which is a prostate-specific membrane probe that goes to tumors in the person. So by selecting the probe, it can be very specific. And also, it shows the function. What it doesn't give you is the anatomy. So you combine it with a CT in order to know where the uptake is going. So one example I would give here that we have been working with, although I will show images from a paper, is the peptides, which is specific for chol-A1. So which is something which show where the lung, in this case, lung fibrosis is building up. So instead of looking at something where the fibrosis is already there, we can look at where the fibrosis is building up. But this is how that works. In the left-hand graph, they gave the probe, but it's a healthy animal, they consider it's no uptake in the lungs. Then they have a gliomycin-exposed animal, which have inflammation and fibrosis going on. And then you consider it's a great uptake. So how do you use such a model in the drug testing? Well, here's one with the antibody therapy. So again, there is the gliomycin with probe, a lot of uptakes. And then they treat with antibody therapy and there is low uptakes. So this is another example of how you can assess the disease. You can, oh, I forgot this one. You can also do this in the human. So here's the CT, you can see the fibrosis. Similarly, the uptake of this compound here. So about PEP, then again, PEP is functional. It's not anatomical. That's one of the main differences. And it also provide all these, not that you wanna have from in the human level, assess disease treatment and so on. And what's the limitation of the nuclear medicine techniques? The radiation dose to the animals here is not really an issue, but the injecting volume is an issue also because you want high activity in to make images, but you can't inject milliliters into small products, which is a challenge that you need very high specific activity. Also, of course, you need the supply of radio-nucleides. Many of them you can buy, if it's a spec, but if it's PEP, you have to be close to someone who produces them. And if you wanna do funny stuff like this probe for Colleen 1, you need a radio chemistry. So that's a challenge. And also the spatial resolution is not that great, but since this is a functional measurement, maybe the resolution is easy. So, look, a lot of good news or good facts about the nuclear medicine, but there's also certain limitations. And then we have the magnetic resonance imaging, which always is a little bit more about the complicated one because you can do so many different things. And it played over the show picture of the mouse heart. We can measure the reaction fraction, but you can really appreciate the soft tissue contrast to get in the abdomen of the rat here, to distinguish the different organs that wouldn't be possible with this way. Or you can use a technique to make angiography of the mouse brain with rather high resolution. And in this particular case, you can see the mouse here going into the magnet. And then I will show some older slides here from my former career. We started inflammatory bowel disease and had a mouse model for that. So this is the colon, the small ring here in a healthy mouse. And this is the colon in a mouse which has this particular disease that we introduced. So instead of work doing, well, we are competing with this. So obviously this is a dead animal because we have the colon outside and we can measure the stiffness, the edema, the bloodiness and the thickness. And that is very operator dependent. So if you want to say, is this animal sick or not sick? Well, it's too late because we have taken it out. So we don't, that's too late to make a study. But that's one thing we can do. We can actually, by using imaging, say, that we have animals with the disease in our groups, which is important. And you can also do contrast and house MRI. So this is now the coronal region of the mouse. So this is the colon. And you can see when we have injected contrast, it's brighter. So, okay, now we found what's over, now it was. So we can actually assess different kind of inflammation in the mouse with acute front and healthy. And what is fun or not fun, what is good is you can do this in humans as well. So it's an incorrect way. So summarize MRI in pre-clinically. You have all these possibilities that we want from a 3D pre-clinical method. What's the limitation with MRI? Well, there is no radiation dose. So that's not an issue. But the spatial resolution, it's really hard to say what it is because you can kind of decide what spatial resolution you want, just by measuring longer time, other equipment, special coils. But if you're not gonna spend too much time on each animal, you're probably in this range here. So if you want higher resolution, it's really long acquisition time. No sensitivity in the cells compared to pets when it comes to looking at different molecules. So ultrasound, I'm not gonna say too much about that, but what differs ultrasound from many other techniques is that they all often have to tell what you're looking at for an audience that are in depth. But this is a beating heart. Which is not healthy. But it gives you the anatomical information and it's direct functional. I mean, cardiology can say how the function works when developing these pictures and they can also do different kind of mission. It's not the technique if you want really high spatial resolution. And again, it's a little operator dependent. And one thing about doing animal is often that you have to shake the animal in order to get good contact with your probe. And that can check how you set up the study because they particularly don't like that. And they don't want to be used for me to know. Okay, optical was shown this morning. The real good thing about the optical is that you can design a very specific probe and put on these molecules that give you in the sense. So that I would say is the benefit of optical. The resolution can be high depending on how you do your measurements. If you want to do in vivo of the whole animal the penetration of the light can also be limitation. So that's a drawback. So to sum up, we hope we have functional imaging. It's the patent specs. They are direct functional, low resolution MRI is indirect functional. You can measure function but not specifically. Spatial resolution is also a limitation. And we have anatomical imaging. We have this excellence of contrast of MRI. Spatial resolution limitation, micro CT gives you anatomical imaging with very high spatial resolution, I would say. But it's also only anatomical data. So if we look on this graph, we have nuclear medicine techniques here with their resolution going down to CT. About this range, 0.01 millimeter. And then we hope to have synchro form out here. So we need experiments that need high spatial resolution, I would say, or something which need to be very, very quick. Sensitivity on contrast media obviously that's packed with our traces. That's sensitive to me. But I guess you can use very many non-aparticles or whatever probe here with in combination with synchro form to make it very sensitive for experiment.