 And we're here in Tessaloniki at the Nanotextology Conference, and who are you? Hi Nicolas, nice to meet you. I'm Antigone Alexandro. I will be presenting a talk, an invited talk tomorrow, on ultra sensitive in vitro diagnostics. So what does that mean? So in vitro diagnostics is to put it in a simple way is what you do when you get blood sample and then you use that blood sample to detect different molecules inside the blood sample and to determine if you have certain different types of disease. So what we are doing is doing the same thing, only we can do it in a much more sensitive way. Something like 1000 more sensitive than standard techniques that are used in a medical analysis lab. So now people use a lot of those glucose meters or something like that, but there's some other things people do also that are much more advanced, like when you do blood tests, right? Yes, exactly. And when they do the blood test, you have a technology potentially to do much more detailed. For example, there are cytokines which are involved in inflammatory diseases and they are so-called biomarkers because they sort of tell you if the disease is there or not. And in many cases, these biomarkers, the current technology is not able to detect them with high enough sensitivity. So the concentrations are there, but they are too low to be detected. And what we propose with our nanoparticle-based technology is to be able to detect these biomarkers, these cytokines, which currently it's not possible to detect them unless you have extremely sophisticated technology. So what do you show here? Okay, so this explains our technology. It's similar to the so-called ELISA technology. What you do is you have antibodies on a surface. Then you incubate your sample, your surface here with the antibodies. You incubate it with the blood sample, which contains different kinds of molecules, which are shown here with different shapes and different colors. Then you rinse and let's say you want to detect cytokine indicative of inflammation. So let's say this is a cytokine. There is a few in your blood sample. So it is recognized by these antibodies, which capture the biomarker. And then you can come and reveal the presence of this biomarker, this green molecule, by another antibody, which will recognize a different part of the same molecule, which is called the capture antibody. Now what you need is a sort of probe to be able to detect the fact that the antibody has bound and the fact that this biomarker is there or was there in your blood sample. And this is where our technology enters. Usually people use techniques that are not very sensitive. But what we propose is to use these lanthanide ion based oxide nanoparticles, which are extremely bright, extremely photo stable. And they give you the possibility to have a very high signal to noise ratio so that you can detect very low concentrations. So if I show you the next slide, this is what you can typically do. There is a notion in this field is the limit of detection. Limit of detection is the lowest concentration you can detect of this given molecule, this given biomarker. And if you use a standard technique that gives you a low signal, if the signal is below this limit of detection, you cannot detect the concentration you are interested in. But in our case what we do, we have a huge increase of the signal due to the very bright probes, luminescence emitting, light emitting probes that we have, the emission that we excite with the laser. And the signal becomes now in our case with our technology much higher than this limit of detection level. And so even a very low concentration becomes detectable. So right now there are clinical studies which use samples and they do the measurements and for certain cytokines, 90% of the samples, they measure nothing. That is, they measure a value that is below the threshold. So they don't know if there is none of the specific cytokines or if the concentration is just too low to detect. And with our technology we will be able now to detect these concentrations and really be able to say if there is some concentration there or not. And in particular we will be able to detect a combination of biomarkers and in that case current molecules that are known to be representative of a disease cannot be used to biomarkers because their concentrations are too low. But now we will be able with this technique which from our laboratory we have commercialized them with a start-up that we co-founded with Max Richley called Lumydix, we will be able to bring new biomarkers to the market because currently biomarkers are there but we cannot detect them. If we are able to detect them there will be new ways of detecting disease more early. So earlier detection means better prognostics for the disease because if you get detected earlier you can treat it earlier and so there is a huge impact that we expect to have with this technology. So these bright dots right here, is it like a new kind of invention that you are able to do this so it's based on established science that's an old way of doing things? Well actually we've been working on this technology with a colleague Thierry Gacouin also working at the Colpol Technique, my institution in Palaisone near Paris. So in my lab and in his lab we have been working on these nanoparticles for about 15 to 20 years now but we've been working on more basic applications like how to label single molecules in the cell membrane and how to track them and see how they diffuse inside the cell membrane and how they are confined in certain cases and how that influences the functioning of the cells. And then at some point we realized that the properties of these particles the fact that they are synthesized easily, they are synthesized in water the fact that they are very photo stable and very stable in dispersion so they don't aggregate. All these properties make them very good candidates for this present application to detect biomarkers in blood or in other human urine samples for example. So how far is this from the market? Actually this startup Lubevix that we co-founded started last year was created last year in July 2018 and we're currently working on the industrialization of the technology and hopefully by next year there will be products in the market. And what are the other things you're working on? You're working on many things or? Well actually we're working with my team together with my colleague Cedric Bouzid. We have a team that is also Nicolas Olivier. We have a team at the laboratory of optics and biosciences in Ecole Polytechnique which is working on let's say as I mentioned earlier understanding cell function and also being able to detect and maybe I can show you another slide being able to detect for example hydrogen peroxide. Hydrogen peroxide, well people used to use it to remove nail polish. Now you don't do it anymore because it's a bit toxic, not highly toxic but you have less toxic solvents that you can use now. Yeah it's just no problem. Maybe you can tell them to speak a bit less loudly. It's just coming right here. And what was known for a long time is that hydrogen peroxide is used by immune cells to kill bacteria so it's a bactericidal activity and you also use it to clean solutions at home sometimes. So what has been found out more recently is that this molecule, hydrogen peroxide is actually also present not only as a toxic molecule but it's also present in very low concentrations, in a very localized manner in time and space and it has a role of a signaling molecule. Now what is signaling for a cell? Signaling is, well cells need to communicate with each other and they need to communicate with the external environment. So there is a signal, there is a molecule coming from the outside. It's detected by so-called receptors on the cell membrane and then there is a transmission of signal so the receptors' role is to detect the signaling molecule and then tell the cell to do something in response to that. For example if you have vascular cells, what their role is to contract for example in response to certain signals. So if these vascular smooth muscle cells, if they detect a molecule called endothelium, this is the signal for the cell to contract. And so the cell has a machinery to transform this outside signal into a cellular response which can be in this case contraction or migration or differentiation or apoptosis. And for that there is an intermediate molecule which is like what is called a cell has a signaling machinery and there is an intermediate molecule. In our case it's also hydrogen peroxide and it's involved in the signaling cascade of detecting the signal from detecting the signal all the way to creating the cell response and in the middle there is this hydrogen peroxide. And what we do is try to understand how this signaling pathway works by detecting this hydrogen peroxide in a very efficient way. Quantitatively we can measure concentrations and resolve in time and in space. And actually it turns out that we use the very same nanoparticles that I mentioned before for ultra sensitive detection. We use these very same particles in a somewhat different situation and the conditions to detect this, to detect with, and so we have a nano sensor. Before we had a label, now we have a nano sensor. And maybe I can show you a slide on this. No, it's perfect. So I can show you how, for example... Is this something that has to do with blood samples also or where do you do this? Actually we do that in cells and that's for basic research so we want to understand what the concentrations of the signaling molecule, what the involved concentrations are and what the time dynamics is. Because we have been able to show with our sensors that the dynamics is different. For example, here is one example. Depending if... Okay, here is our molecule that activates the receptor and then this receptor leads to production of hydrogen peroxide, this signaling molecule that I was mentioning before. And depending on the type of the stimulation of the cell, this is a molecule inducing contraction and here this is a molecule plate-blade-derived growth factor that induces migration. And you can see here that the time scale, the dynamics, the time dynamics, is very different between these two stimulations. So the secondary messenger that we use, that the cell uses is the same, hydrogen peroxide, but the time dynamics is different and the concentration is similar but the time dynamics is different and we are, thanks to this nano sensor based on European ions, we were the first to be able to detect this. And now the next step... So what do you detect? What can we do? Well, we detect as before the luminescence of these nanoparticles. The only difference is that before we would use them. So we remove electrons and then this molecule has oxidative properties. Sorry, we add electrons and this molecule has oxidative properties so it removes electrons from the nanoparticles. So when we add electrons, I don't show it here, but what happens is that the luminescence signal decreases. Then we stimulate the cell and then hydrogen peroxide is produced. Here are the nanoparticles we... Oops, I'm sorry, which we have managed to put inside the cell by relatively standard technique and here is the white light image of the cell and here are the nanoparticles emitting signal luminescence inside the cell. So we follow the luminescence of one single nanoparticle. So the luminescence goes down when we add electrons and then when we stimulate the cell we produce this signaling molecule and then it oxidizes the nanoparticles back to their initial state and then we have a rise in luminescence. And from this rise we can extract very precisely, quantitatively, and time and space results. Time result because you can see we can measure the concentration of hydrogen peroxide as a function of time but we can also measure locally because each nanoparticle, here, each single nanoparticle that you can visualize with a high-performance luminescence microscope gives you the information of the hydrogen peroxide in the very specific location of the cell where the nanoparticle is located. So we have time resolution, space resolution and quantitative information. So what could this possibly help with in the future? Okay, so what I've been saying is more trying to understand how the cells work but it's one very important issue in basic science is you never know what application it will lead to so it's very important to have basic science if you want to have applied science later down on the way and so here is the application. And this is relevant to many different materials, many different particles? Actually, for now, this is the only nanoparticle that we have identified that has all these multiple properties localized and time result detection and quantitative detection. Other nanosensors don't have all these combination, this combination of properties and the next thing, so I've told you this is what we call physiological signaling what happens when the hell is healthy but then this process can get deregulated and then you get deregulated hydrogen peroxide production you get what we call oxidative stress you've probably heard about that it's often mentioned in everyday life about oxidative stress and you have to use antioxidant to remove the oxidative stress it has entered everyday commercials and then you can have if you have deregulation of this hydrogen peroxide production you can have neurodegenerative diseases you can have inflammation, chronic inflammation which is related to cancer and many diseases which are related to inflammation like diabetes or as I said neurodegenerative diseases and so the next step is to try to do this detection in vivo, in vivo meaning in living organisms so we start with mice and so this has not been published yet but what we did was of course there's an ethical committee that validated this research program beforehand we did that with Gustave Roussi which is a hospital which is one of the most well-known cancer hospitals and research centers in France and in Europe and what we do is we have a mouse model of inflammation so we inject the particles in the mouse ear and then we apply a molecule called methyl salicylate which induces inflammation of the ear actually it's what also some people use in sports to heat up their muscles okay in this case you generate an inflammation and here, okay we can see here in this image you can see the particles the emission of the particles that have been injected in the mouse ear and then when you do this okay first we check that we indeed have an inflammation with other standard techniques I will skip this part and then as I showed you here this is the signal of the nanoparticles so I will start the movie now and when I start the movie you will see in the beginning nothing happens the signal, the luminescence signal oops, I'm sorry let's just go back so in the beginning nothing happens then okay I will go back because it goes a bit fast oops, I'm sorry yes, so in the beginning nothing happens then we apply the induction of inflammation so that's when the mouse ear shifts a bit and then if we wait a little we see the increase of the signal that I told you which reflects the production of hydrogen peroxide and other oxidant molecules so here again it's one of the first measurements of this oxidative response which is a pathological condition especially if it becomes chronic and so we have been able to measure as a function of time and for different mouse ears the increase of signal and how it is and from that we can extract the production of hydrogen peroxide so if you remember what I showed you before here the signal is I didn't show it actually but the signal here is much higher than before because the concentrations and the signal related signals for physiological mechanism is much lower here we have a pathological mechanism it's an inflammation it's a reaction of the body to this application of an external molecule and we have a higher signal higher production of hydrogen peroxide so ideally we would like in the long term to be able to do this in humans but this will still require some years of work to come and so these signals and detecting those signals can be useful for many things for yes, as I mentioned there are several pathologies and there are many pathologies where inflammation is involved cardiovascular disease is also involved as an inflammatory response neurodegenerative diseases cancer, many cancer types are related to chronic inflammation like for example the gastric cancer comes from ulcers and ulcer comes from helicobacter pylori bacterium it creates ulcers it creates a chronic inflammation and if nothing is done the chronic inflammation can turn into cancer so all these diseases it's important for all these diseases it's important to be able to measure as many parameters as possible of the disease to be able to do what is called precision or personalized medicine because, okay currently you measure one or two biomarkers but if you can also measure what is the concentration of hydrogen peroxide for a certain patient and how that correlates with the severity of the cancer or with the metastatic activity of the cancer or you can correlate it that we hope to do in the future how it correlates with the efficacy of a certain pharmaceutical treatment so this tool measuring the reactive oxygen species that are produced of which one of the main ones is the hydrogen peroxide this can give a valuable new parameter for a huge number of pathological situations so you can get an injection of something and then all the cancer cells will light up or how does it work well we are currently starting to work on that we want to avoid injecting, we want to avoid injecting the nanoparticles in the human body because there are always toxicity issues so we would like to be able to just introduce the particles do the measurement and then remove them so maybe in the next conference I can tell you more about this and you in Paris, right? Yes, I work in south of Paris at the Ecole Polytechnique which has now merged with four other institutions to form the Institut Polytechnique de Paris and many students in your group or how does it work? Yeah, we are three permanent researchers one technical staff and graduate students postdocs and undergraduate students so typically a group of about 10 people but we are a part of a bigger lab with 50, 60 people the laboratory for optics and biosciences and our team is called nano imaging and nanotechnology is a big deal yes, nano imaging and quantitative biology yes all these nano tools are now almost everywhere in all aspects of life and of course we have to make sure that we use them only where they are really necessary so that the public does not develop Nano addiction? Not nano addiction but an apprehension of these technologies in some cases they are used in sunscreens for example in some cases they are not really necessary so scientists have to be really careful to in terms with interaction with society have to be justified why we use the nanoparticles and that we use them only when they are really necessary because of course there are also human toxicity issue but also toxicity issues to the environment are used at a massive scale and if they are released in the environment you can have issues like we have today with plastic maybe if we don't pay attention we'll have similar issues with nanoparticles so hopefully not but scientists in this field have to be very cautious about issues like that so you'll have a presentation about this tomorrow do you have lots of networking with people from nanotechs? this is really a great occasion to meet very exciting people in different fields in my field of nano biomedicine and nano bio biology but also to see some very exciting plenary talks on other fields of nano sciences so I'm really delighted to be here and I thank the organizers for inviting me