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Intro/Outro (00:00:02):
Welcome to The Microscopists, a Bitesize Bio podcast hosted by Peter O'Toole, sponsored by Zeiss Microscopy. Today on The Microscopists.

Peter O'Toole (00:00:19):
Today on The Microscopists I'm joined by Markus Sauer of the University of Wurtzburg. And today we discuss things from the study of combustion to studying single molecules.

Markus Sauer (00:00:33):
Very all of us in, in this, in this department have been working on combustion and I started to focus now on biological molecules and imaging in microscopy and single molecule techniques/ Without any knowledge. So it was a one man show in a physics department basically.

Peter O'Toole (00:00:54):
Why his winter meeting always starts at 4:00 PM just to allow for a day skiing or is it?

Markus Sauer (00:01:02):
Uh, we are always looking forward to, to meet in closeness every year for skiing and, and, and In science exchange.

Peter O'Toole (00:01:10):
His attempt to get us to eat our vegetables.

Markus Sauer (00:01:15):
There are a few vegetables, I didn't like that much. It's not that I hate vegetables. I eat vegetables on a daily basis

Peter O'Toole (00:01:23):
Where he thinks further research is needed.

Markus Sauer (00:01:27):
We have to find ways to efficiently label sites and proteins with super small displacement.

Peter O'Toole (00:01:39):
All in this episode of The Microscopists.

Peter O'Toole (00:01:47):
Hi, I'm Peter O'Toole from the University of York and today on The Microscopists, I'm joined by Marcus Sauer from the University of Wurtzburg Germany. Marcus, how are you today?

Markus Sauer (00:01:57):
Good. Thank you very much. How are you?

Peter O'Toole (00:02:00):
I'm I'm really good. Thank you. And thank you so much for accepting this invitation and joining me today. I remember first. I remember the first time he, hearing you talk which was at an ELMI meeting and I was blown away by your research, but I was also, what was really impressive. Yeah. We've read the papers. We know the research. We kind of know the field. What was really good is I'm listening, criticizing and get in my head thinking. Oh yes. But what if, and then within two slides you had the what if it's like, oh, he's answered it. I don't think, oh yes. But what if two slides later? Boomf, that's it? It's like, oh my goodness. I think I can say, ah, yes, this all sounds great, but, and then you say, well, this might sound great, but look at this. So actually we can't believe it, but it really we've done it. Oh yeah. I was really taken aback by how comprehensive, how you criticized your own work and addressed it in the presentation. And that was a talk, I think you were counting around a nuclear pool and counting the rings or similar to that, which was immense. So that doesn't come naturally that skill. So where did you start your career? What was your first degree?

Markus Sauer (00:03:29):
Where did I start my career? First degree graduate school in Pforzheim. That's hidden city Southwest between Stuttgart and Karlsruhe. This is where I'm growing up there. I did my graduate school and well and I, I, I graduated 1984 and if I remember correctly, I even de-select it from my courses physics and chemistry at the end. Yeah. So you could select courses where it would put more focus on or less. And I de-select the chemistry and physics at the end because I thought it's super boring. And on the other hand also quite complicated. So I de selected that and well, but simultaneously I was very fascinated by medicine and how it could be possible to, to manage or heal diseases yeah. Or treat cancer, or find new drugs or new methods to, to help people and to fight against some of these big problems in medicine and things like that. And, and this is also what, after, after I graduated, I also tried to, or started actually to study medicine for two semesters for about one year. And if you are studying medicine, you have to work in hospitals, kind of practical seminar, practic, practical courses, and things like that you have to do. And while I did that there was a professor of physical chemistry professor in the hospital. And I told him about my dreams, what, what, what is fascinating me and what I would love to do later and things like that. And he told me, Markus, if you're really want to do that with your talk, telling me here do not, you should not study medicine. You should study physics or chemistry and then develop new technologies methods, which have a chance to be translationally applied in medicine in a way to help people and to fight against disease or to find new drugs. And this really changed my mind. It was mind blowing me. And because I always also was quite frustrated during the first two semesters medicine, because it was like a memorizing course to learn, learn, learn. And then next day people ask you and get to remember and, and give exactly what you're memorized. I mean, this was also not what I aimed for. So I started then to study chemistry. And so this was the start of everything to start chemistry. And then after one of studying chemistry, I was a bit bored about all these chemical formulas. So it focused more and more into the direction of physical. Chemistry. And then a very important step was then that I ended up in a group. So my PhD supervisor Jürgen Wolfrum was, is his name. He's still alive 80, 82 years old now. Exactly. And he was doing combustion diagnostics. So he was using lasers and investigating diesel or gasoline engines so that the combustion process yep. Doing imaging of radicals, which are produced during combustion and the distribution and how to optimize that and things like that. So it was one of the leading persons in the field. And, and I thought that's interesting lasers and imaging, and it's a bit of microscopy and it's photo physics of radicals and for photochemistry. And so I started my PhD in his group. And after one year of doing PhD in his group, he came back from a trip to US where he was, I think he was in the advisory board of the us government that time. It was when was that? 1991 92, something like that to put, or to develop lasers for defense system in space. I remember something called STI project or something. Yeah. And he was on board of that. And while he was there in us for such a meeting he, he met just by chance. I think Rex Keller, maybe this name doesn't tell you anything, but he was, he was at Los Alamos. He was head of the group responsible for seeing the molecule DNA sequencing. In early times already in the eighties, they started projects to develop or find ways to sequence a single DNA molecule. And he came back and told me super fascinated about the ideas of the Keller and told me, Markus, you have to write a grant in it to start doing research this direction. And this was finally the beginning of my way to, I mean, I was, I was the XO, the very, all of us in this, in this department have been working on combustion. And I started to focus now on biological molecules and imaging and microscopy and single molecule techniques without any knowledge. So it was a one man show in a physics department. Basically.

Peter O'Toole (00:09:02):
I'm just going right back to the very start where you found chemistry and physics, not so interesting, boring and not so challenging. So you went into biomedicine, you then get told by a, quite an insightful medic that you should do chemistry and physics. If you want to make a difference that those impacts that you wanted to do. So not only did you go into chemistry and physics, but you find yourself in a combustion engine group, which has nothing to do with human research, nothing at all. And then somehow you start working on DNA side of combustion engine

Markus Sauer (00:09:37):
DNA. So he pushed me to, to try to find ways to detect single molecules by fluorescence. And, and I tried to find literature and there wasn't much literature at that time. So it was basically two or three papers from Los Alamos from the Keller's group. They tried to do that in a flow system. Yeah. And then there was a connection between Heidelberg where I did my PhD and Göttingen and to, to the group of Manfred Eigen and Rudolph Rigler. And it was the same time they started to,uto, to, to use FCS fluorescence correlation spectroscopy in confocal microscopes to increase the signal to background ratio, to be able to detect fluctuations at the same molecule level in the first signal. Well, finally, we're joined forces between Göttingen and, and Heidelberg. And this is, was also the first time when I met Petra Schwille

Peter O'Toole (00:10:38):
Petra was right at the forefront of,

Markus Sauer (00:10:40):
Yeah, she was at the forefront of this FCS business with Manfred Eigen and Rudolph Rigler. And I was a one-man show in Heidelberg. And for some reasons we joined forces and, and I started, which was then my speciality to, to learn about the photophysics and how of dyes and how to, to which dye you have to use for which application. And, and also things like, can you use photophysics advantageously to, to monitor interactions of thighs with biomolecules and things like that. Then this was then my specialty. U

Peter O'Toole (00:11:21):
Markus actually the one question I have for you talking to Petra. Don't you go skiing once a year with Petra.

Markus Sauer (00:11:30):
Yeah, well we are going skiing once a year in general, early of January, there is a a conference called the winter seminar. It has been originally introduced by Manfred Eigen, our Nobel Laureate from Göttingen in the late sixties as a kind of group seminar, the Alps,

Peter O'Toole (00:11:54):
I presume this isn't the group.

Markus Sauer (00:11:57):
Oh, this isn't the group. No, this is my group. And at the, I think it was 2008 or 2009. I have been asked whether I would like to take over and organize the seminar together with a colleague from Berlin. And I finally agreed to organize the seminar. And since then the last 12 years or 11 years, I'm organizing this, this annual winter seminar in the Alps January it's hold in, Switzerland in Klosters close to Davos uin a hotel. And then we offer 10 to 14 days every January and Petra is coming regularly. I mean, she did a PhD already, Manfred Eigen school. I met her there every year. I'm meeting other colleagues there every year, more or less. We always invite people from around the globe. Just, I haven't had an invite yet. It's a very private format. I mean, we can do nothing but webpage. I mean, maybe we should do that. So there is about 150 people coming per year, but they do not all stay for two weeks. So I stay for two weeks. That's good for me. Yeah, exactly. I go skiing with friends, with colleagues and so on. So these are a few colleagues in the background there. I see a few colleagues from,umost of them are on Michael Groll from Munich is there. And,uMike Heilemann from Frankfurt, I see in the background and a few people who regularly join,uthe seminar, but also two older,ucolleagues older, sorry,ufrom, from Göttingen. And so this is,uwhat we are always looking forward to, to meet in closeness every year,ufor skiing and in science exchange. So it's, it's, it's a very,ua very easy format. So we have basically two to three, four talks starting at 4:00 PM. So there's enough time for skiing the whole day. And,uthen a few talks and, and drinks at the bar and quite relaxed atmosphere,

Peter O'Toole (00:14:19):
I would say this for anyone who's not into conferences. Most conferences are very intense periods of silence. Right now you're selling this two to four hours of seminars. And I think I can see exactly why this is a good conference and it's not, and it's not because of the social I'm presuming the, the free, the free mind talk, the ability to think you have two or three big lectures, big, you know, they

Markus Sauer (00:14:53):
Are really big. They're really big. They are one hour each with 15, 20, sometimes 30 minutes of discussion about it afterwards. And

Peter O'Toole (00:15:02):
Then in the bar, I'd imagine there's a lot more talk about the science,

Markus Sauer (00:15:07):
But I mean, if you want to start a cooperation, I mean, there is nothing better than going skiing with the person for one day.

Peter O'Toole (00:15:14):
Yeah. Yeah. Do you know, this is you're in the zoom world as we are today. It's great. And the existing networks exist, but to create networks is really tough. You know, it's a bit like any conference it's down the bar when you're eating that you make new contacts that you can talk more freely unguarded and come out with nonsense ideas, but those nonsense ideas, and you can

Markus Sauer (00:15:44):
Also address the questions you, you do. Usually not address after a talk. Maybe more critical questions are just, maybe you think your question might be too stupid, so you don't ask it after the talk, but you might ask after a beer or during a beer or so. Yeah. I mean, this is the advantage of these conferences. And then I have another ski seminar in March, usually every year with my group, but only for one week, that's it to, who's the best skier. Ah, the guy orange vest in the orange. He's a professional free ride skier, but he left the group a year ago, but I learned a lot of him from him in the last five years because he's really professional doing free rides skiing. And wow. I mean, I was a good skier, but I didn't do that much free ride skiing, but thanks to him, a few members, including me, we learned free ride skiing with avalanche rucksack and everything, fully equipped to do a few things, which are really scary, but fascinating if you so love

Peter O'Toole (00:16:54):
You'd like your so you like the thrill, something scary as well. These things, I

Markus Sauer (00:16:58):
Really love, not, not all scary thing, things. I mean, there are other things I don't do. Yeah. but skiing free ride somewhere in the Alps. This can be also, doesn't have to be too dangerous. I mean, you are equipped with protectors with with this with this avalanches on your back and things like that. So, I mean, you're not stupid. Yeah. But it gives you a kind of freedom.

Peter O'Toole (00:17:30):
So, and in that, so that's great. So you get a thrill outside of work with your skiing in the winter. What about the summer?

Markus Sauer (00:17:37):
What about the summer relevant motorbikes?

Peter O'Toole (00:17:44):
I've got to say that is an awesome looking, British racing green,

Markus Sauer (00:17:51):
Exactly. British racing. Green Triumph. I mean, I had motorbikes when I was 18, 19 till 22 or so I was riding motorbikes, but I never could afford to buy a Triumph. Yeah. but then, I mean, I stopped motorbiking for 30 years, almost 25 years because my wife thought it's too dangerous. But then I convinced her that in order to ride in routes, work between the different institutes without parking lot slots. Yeah. Without parking places, I need a motorbike again. And finally I could convince her to buy. First of all, I bought a smaller one. And then I could realize my dream of riding a triumph for special design and everything. Yeah.

Peter O'Toole (00:18:35):
I, I'm not, I'm not, I'm not a motorcyclist, but that does look really, really gorgeous. It was

Markus Sauer (00:18:42):
A special design with all Catholic Whitman and blah, blah, blah. Well, now I am not a rich person, but I think now it's much easier to realize a few dreams you had when you had been 20 years old. Yeah. And otherwise

Markus Sauer (00:18:59):
I love the fact that your wife didn't let you do it too dangerous. And up to 25 years of marriage, she said, she ain't go for it, please.

Markus Sauer (00:19:11):
No, that was not what she said, As I told you, my argument was to drive too, because I started with several collaborations, but two medical doctors at the university hospital. And in order to commute between my Institute and the university hospital, it's much easier if you either go by bike, but let's book is very hilly. So no way without an e-bike at least, or you buy them buy motorbike,

Peter O'Toole (00:19:37):
But that leads us back into work quite nicely. Because again, you still have to be with that chemistry, physics, not liking it, then going back into chemistry, physics, and then you come back full cycle. And now you are working with the medics, you're working, collaborating with the research. So what, what are they doing with your techniques? What, what are you bring into them that they couldn't otherwise do?

Markus Sauer (00:20:01):
I mean, it's, I mean, there have been, I don't know how many super-resolution microscopy techniques developed very successfully and they produce impressive results in biological research. Like you mentioned, though, we talked just briefly about nuclear pore complex is to, to, to unravel that the molecular architecture of multi protein complexes and things like that with no-one, nobody's succeeded so far to really translate these technologies or use them advantageously in, in medical sciences. And, and I think it's started here 5, 6, 7 years ago when a discussion with the director of the university of hospital of University Hospital of Würzburg. [Inaudible] And he, we were in discussion about immunotherapies, and he's developing immunotherapies to treat multiple myeloma and leukemia patients is a international expert on that. And he told me that he told me a few stories about, so they address tumor associated antigens on the surface of these tumor cells. So they are attacked by antibodies, for instance, immunotherapy, with antibodies or what they do today. So-called car T cell therapies that they take the T-cells out of your body. And then they genetically manipulate the cells that they expressed a few binders on the surface, which are directed against these tumor associated antigens. But he totally about side effects of this therapy that some people pass the pass away under this immunotherapy. And they don't know why and people discuss about the possibility, but that these receptors might also be expressed albeit at low expression level on healthy cells and things like that. And this is when I, it there, we can help me to stay motivated, sensitive imaging of tumor cells and of healthy cells and label these receptors with antibodies and tells you something about the number of basic, how many receptors are expressed. And this is what we are doing now in, in, in several big approaches together with university hospital here and in Berlin [inaudible] and other people to use this super-resolution and single-mode techniques too. Yeah. Pump a little bit to try to improve this immunotherapy P applications.

Peter O'Toole (00:22:26):
I've got a question that would be really interesting to hear. How did you find this group? How did they find you? Where did you meet

Markus Sauer (00:22:37):
The director of the university hospital? I found here, I met here at the meeting of the university. I think I gave a talk and he gave a talk. I think there wasn't, it was just by chance, I mean, this is always chance just by chance. You, you have to meet somewhere where you have to be open-minded. I mean, this is the major, problem. The people, I mean, yeah. Usually you might leave the room and grab a coffee. If a medical doctor is giving a talk after you talked. Yeah. But maybe if you stay in and then start discussions afterwards with him and he listened to your talk, then there might be possibilities to bring these two sides together efficiently. And this is really fascinating me a lot, I have to say. So this is one side of my fascination to really use this, these technologies and these microscopes yeah. Patience on patient samples. Yeah. And then the other thing is still photophysics of dyes and how to use that to, I don't know, to understand a few things. Yeah.

Peter O'Toole (00:23:43):
I think that's yeah, really important. And it highlights it going back to the skiing trips and the meetings and the conferences. It does bring people together from different disciplines. There's always going to be someone who's a bit different outside the field, right? It's those meetings and the relaxed atmosphere that enables those connections you made, but you're right. You stayed in you listen to a talk that wasn't relevant on paper wasn't relevant, but when you listen suddenly it's like, well, actually I can contribute to that. But if you weren't at that meeting, you wouldn't have picked up his scientific paper. I

Markus Sauer (00:24:18):
Wouldn't never, ever, never, ever

Peter O'Toole (00:24:21):
The importance of those meetings. It's really nicely exemplified by how you, how you met so to speak. It's quite nice that that, that came through on that. So talking about going back to the, the more, the more fundamental research I had a question from earlier that I didn't ask how much fluorescence is there inside a combustion engine.

Markus Sauer (00:24:41):
There is a lot of florescence. These are radicals O H radicals, nitrogen, oxygen radicals, and the distribution of these radicals tells you something about the, the exhaust how many nitrogen gases and, and, and other gases you do not want to have today are exhausted how to prevent it.

Peter O'Toole (00:25:06):
So is that, that multidisciplinary approach, just what we can learn from this assist the engineers, material scientists

Markus Sauer (00:25:14):
Slowly, this, yeah, this was super multidisciplinary surrounding that time. So my colleagues, I mean, I jumped out of this combustion business pretty fast, but my colleagues, they moved with lasers and, and equipment to Mercedes in Stuttgart or to Peugeot in Paris in Paris. Yeah. To do measurements at their location in Paris, on their engines there, and try to tell them something how to improve the combustion process. So this was very, very interesting, I think, but it hasn't had nothing to do with biology or medicine. Yeah. So this, this took me really a hard time at the beginning. I mean, imagine you're working alone in a field and nobody else is working. Nobody can help you. You can already ask for advice or, or help. I mean, there was one person at [Inaudible]. He was, he's not 88 years old, I think, but still alive. Um and he was in, he lives and he was at the university of Siegen in Germany, but he was he's did really seminal work on, through for development. He worked for Kodak in US for example, and develop most of the Rhodamine we know today in use today, he was involved in the development of these dyes and many other things nearfield interactions. He did seminal work and he was my Co course supervisor of my PhD. And I could learn enormous. And I learned all I know about Fluorophores in dyes. I Learned From Him And There Is Still A still people are reading. If you read today old books from [Inaudible] of the seventies of last century. And I promise you will learn a lot of things. This is a secret tip, read the very old papers of [Inaudible]. And you, you will learn everything about triplets states and quantum yields and why the quantum yield is higher for this dye or lower for that dye and crunching processes and possibilities for quenching. This is really fascinating. I could learn a lot from it.

Peter O'Toole (00:27:28):
I said, now I have to go back and read that third promise I've made on these podcasts. I think I promised Richard Henderson a lot breaking bad, and I've started Elizabeth Hellman. I'd probably start cleaning my oven and I've done that. That's ticked off now. Cool. So now I got to read more about fluorescence nd I thought flourescence was pretty good, but actually I would love to get deeper with really, so I will take you up on that. I will go for that. My next question is what was the first microscope you used?

Markus Sauer (00:28:01):
Yeah, well, the first microscope was a microscope. I built myself during my PhD. I mean, my, my idea had been, or I was told to detect the fluorescence of a single molecule. And there was no paper published about that by confocal microscopy or what, what is standard today? A widefield microscopy. I mean, I found a launch for the dyer in the lab an objective water immersion objective with a high end A and so I put a drop of water with dyes diluted to ten to minus 10, 10 to minus 11 molar, put it onto the objective lens placed the other loss. Photodiode in the imaging plane of the oven of the objective, and then irradiated from the site with a lens. I focus the laser into the droplet, into the water droplet. And I saw on the APD some fluctuations. And I didn't know what it is that it was my first thing market signals. I'm pretty sure you just do the [inaudible] meter, but then due to the cooperation with Göttingen, we learned from Rudolph Rigler that you should better take a confocal microscopes. So the confocal principle, and I mean, we immediately, I immediately tried that and worked out much better. Yeah. So, but I re I will never forget these times when we aligned the first confocal microscopes to detecting molecule signals. Why, why did they transfer through the laser, why they pass through the laser-focus? And we were sitting until 5:00 AM in the morning in front of all the monitors and seeing some fluctuating signals, but you never knew what is it, is this the signature of seeing molecules or not? And then you try to sleep and to close your eyes, and you still saw all these fluctuating signals and today it's so easy. It's so super easy today.

Peter O'Toole (00:30:10):
Don't you miss it from time

Markus Sauer (00:30:13):
To time I'm missing it? Yeah. I mean, I haven't been in the lab for 10 years or so. I would destroy everything probably or misalign or I don't know

Peter O'Toole (00:30:27):
But that Unknown, you know, not know what the signals even mean and prove and get that that's or increase the concentration, dilute the concentration.

Markus Sauer (00:30:35):
Sure, sure. That's easy. I mean, for example, I think I'm pretty sure that we monitored and recorded for weeks, the signal impurities in the oil of an oil immersion objective without knowing it. I'm pretty sure many, many, many scientists did that in the early years.

Peter O'Toole (00:31:00):
Oh yeah. Okay. Moving on. So that was your early days, but obviously you've made significant impacts moving forward. I will dare to say maybe dStorm being one of the biggest to differ. So yeah,

Markus Sauer (00:31:15):
But this was, this was a difficult story. This time was really a difficult story. I mean, I, I was in the years of 2000 to 2005, my work mainly focused on phot physics of dyes at the single molecule level to investigate flora for interactions and blink behavior of dyes and with the aim to use it maybe at the end for the data storage at the single molecule level. So these have been the things we have been dreaming of. Yeah. And then we discovered just by chance that this copper signing dyes scifive and Alexa 6, 4 7 that they show this kind of addressable reversible blinking in some real buffers. But these buffers were not that weird. I mean, there have been papers published in nature already 1990, 1988 or something already by, by by Yanagida, a Japanese scientist. But he was very popular and famous in in the last century for his work on ATP, a single molecule rotation measurements and tracking of Meyers in molecules and things like that. So the first thing, molecule measurements, and he always used designing dyes in an order to improve the photo stability. He used files and a captor methylamine or my captor methanol. Yup. He introduced that. And then there came the time of a single no Fret measurements. So this was the time of human rights and take CIPA. When I was in shamans lab, I was there in 19 8 97. It was just the last few months of take Japan. And yeah. So where the overlap of a few months there, I will never forget. And there we did this first thing, molecule Fret experiments, and there people also use this methylamine or methanol buffer. And from time to time, people reported then afterwards that they saw a strange behavior in the Fret efficiency so that the fret efficiency seems to be high. And then it drops for a few seconds to zero and then it recovered back disappears completely. And then it could recover spec if you monitor trajectories. So seeing him immobilized through force, and this we, we, we w we thought about let's investigate this in more detail. And this is how we discovered that these dyes can be reversibly for, to switch by means of light and fire buffer between on and off. And then we published this, but not for localization microscopy, just for, I mean, just for scientific reasons. I mean, this is maybe a way to, to, to do data storage at the single molecule level on, off, and everything the same when we publish this paper,uuwe send it to science, but they rejected it and it ended up then in Jacs anyway. And in the same week, when this paper appeared in Jacs, there was this other paper from Harvard uwhich appeared in PRL and they discovered exactly the same. And they described identical results. Yeah. The only difference was we publish this for site five, how scifive can be switched. And the other piece of paper,upublished that agreeing for, for activator is required in order to get scifive switched. So, okay. We thought about it. Okay. No explanation for that. Why maybe they also listened to this talk of the fret people and they just took the fret pair to investigate this in more detail. Yeah. I mean, this would make sense to me anyway. And then of course it was the, the, the, the work of Harvard and [inaudible] and the people in Harvard to then,uuse it for the first time advantageously for storm microscopy. There is no question. This was their Uand, and, and justified fully justified. Yeah. Everything. But I, I met then in these years shall why several times. And I told her, I mean, it's much easier why you do not have to use this activator shower. Why, why aren't you using it? I mean, we published this in the same week, in another journal. Yeah. That they were ignorant. I don't know. It was kind of ignorance. No, no. We do this complicated labeling of antibodies with the green and the red dye. And this pushed us to the point that we said, meanwhile, I I'm grateful for that. Then we said, okay, then we'd publish our own work. And in order to honor this work of Harvard, we didn't give it another acronym. We just added a d because we thought that's quite fair considering these two papers, which appeared simultaneously, but this brought a lot of problems with Harvard, right? This is, was not so nice that the following years then, because of this fight between,udstorm and storm and anyway, but we all survived.

Peter O'Toole (00:36:30):
But from an end user perspective.

Markus Sauer (00:36:33):
Yeah, yeah. It has always been my argument, but it was delicate. That is caching and delicate that time.

Peter O'Toole (00:36:43):
It's interesting to hear that the politics you know, you're not in the same groups, you're competing scientists, but you still talk to them. You, you didn't do it without knowledge.

Markus Sauer (00:36:59):
We, we, we, we, we, we, we took a technique developed by others, but this was not the truth. I mean, a PALM, a microscopy, or for the activated localization, microscopy has been invented at the same time much earlier. If you have a look into the submission dates of these papers, but anyway, I mean, mean while everything is super relaxed and cool, but the first years after that, because there was a rush, it was clear that there might be a Nobel prize given into this direction. It was not so nice. The situations in meetings, there were fights, there were fights. It was the time when fight started between Eric Betzig and me and there were everybody tried to claim something. Yeah. It was interesting.

Peter O'Toole (00:37:47):
I think the field, I think it's amazing for people who are not familiar to see that these politics and they were fights, so they're quite public, but yeah, that were going on. I think that's less. So the case today I think the film is more collegial maybe. And,

Markus Sauer (00:38:06):
And do you guys do it, I'm really happy for the fact that the Nobel prize has been given. I was in the first four, four super-resolution microscopy in the first, second. I was really shocked about it. I thought, oh, that's too early. Yeah. It's much too early. What happened? Yeah. But finally, I think it was a good thing because, because it made super resolution microscopy even more prominent and known. And so we also profited from took it. Yeah. It helped

Peter O'Toole (00:38:33):
That. And I think putting the letter at the start a storm has been followed, but in many times overnight by different techniques, putting adding letters to, to the, the original acronym.

Markus Sauer (00:38:42):
Yeah, exactly. Yeah. This was a same molecule, active control microscope lobster. I really know a few of these acronyms. Very cool. It's really cool time. I have to say. Yeah, but very powerful technique at the end, but was then DNA paint. I have to say, this is very powerful, restricted to turf microscopy, but it's super powerful. And it definitely achieves a higher resolution than the storm technologies.

Peter O'Toole (00:39:14):
And that that's where the future goes. Because those as we, most of these techniques are limited in one play or small defined space and it opens up over time. So you mentioned as well that you sent it to science. And then so you went to Jacs, which is a very good journal, but you know, obviously it's a high very high impacting publication in the longer term. How easy do you find your publishing?

Markus Sauer (00:39:41):
This is still still something I do not completely understand. I mean, from time to time, I mean, just historically speaking in my last, 20, 25 years when I wrote a paper and I was super fascinated and thought this, science or nature. It never ended up there it never ended up there. I got it immediately back. And with editor's opinion, this is not new. This is boring. This is whatsoever. But others, other papers where I personally thought, well, it's good, but you could publish it in a nature journal. Yeah. Without any problem, so this is unpredictable, this is super unpredictable. It depends on the personal opinion of the editors. And then of course, on the reviews. But I think I do not have to complain.

Peter O'Toole (00:40:50):
I think it's, it's Not a lot of me, but it is.

Markus Sauer (00:40:52):
It's not a lot. Exactly. This is not what I wanted to say. That it's a lottery. Good luck. Yeah.

Peter O'Toole (00:41:02):
My time. Yeah. Yeah.

Markus Sauer (00:41:05):
Yeah. There's also a saying that says, if you publish really excellent work in the lower impact journal, it will be seen by everybody as well. And

Peter O'Toole (00:41:17):
Increasingly, so it's easier and easier to, to, it doesn't make to a degree it's less important where it is published, but I think universally people still pride themselves on where it gets published. I think that aspiration is good because it keeps people and it makes their sites to be as excellent and as adventurous and as impactful as possible. So it's not a bad thing to Rita Strack, one of the podcasts too, really nicely about Nature methods and the challenges of saying no or accepting it and the fun and the under the, the heart from their side, it's pretty tough.

Markus Sauer (00:41:55):
It's Super fast. So, so, so, so I know Rita, well, dare I say very well, but I mean, we're in contact from time to time. And, and I mean, I see that they are doing really complicated job yeah, which gives them a hard time sometimes to reject papers. Yeah, no, I, I always accepted these decisions.

Peter O'Toole (00:42:24):
So what is your favorite publication that you've ever authored or co-authored?

Markus Sauer (00:42:31):
Well, this is very difficult, but I think I'm still fascinated by phot physics of dyes. And there have been some photo physics papers which we published in the years, 2000 to 2005, including this first photo switching paper or Scifive, which I'm still, I think these were where the best papers published from my group personal opinion, of course, personal opinion that these don't things nice. Super nice yet. But for example, what I'm really I'm not angry about it, but I mean, the nuclear pore complexes we published the first paper on the storm on nuclear pore complexes, 2011 or 2012. And it was really nice, fantastic images we produced about the eightfold symmetry of this nuclear pore complex. Wonderful. But I didn't even submit the work to science or nature. I thought, I mean, there is nothing new we learned zero nothing. So let's submit it to a medium impact journal anyways, kind of strange to see afterwards that people could publish nuclear pore complexes two years later by storm dstorm in science. So this was then frustrating. Not only me, mainly my students who did the work. Yeah.

Peter O'Toole (00:44:01):
Yeah. Cause of course it makes a big difference to their future careers. You know,

Markus Sauer (00:44:06):
Exactly. It's a, it's, it's a problem and they are suffering from these rejection things. And there are also papers. You do, you, you fight to publish for two years, you send it from journal to journal and I'm used to that. Or we all experienced scientists all used to that, but for, but for PhD students, I mean, this is clearly frustrating them.

Peter O'Toole (00:44:30):
I only some quick, quick fire questions. Okay. So what's your pet hate in life? You can't say podcasts.

Markus Sauer (00:44:42):
No. No. Hate, if people are telling you the truth, if they just tell the only, the positive sides of stories just in order to, to, to receive more attention or I don't know, to get for scientists to get people, to get papers sooner published or more efficiently published. But which, which are finally of no help for the audience or for the community. Because useless, nobody can cook it again. Yeah. This is what I really hate. Things like that. Okay.

Peter O'Toole (00:45:29):
What is your, what is your favorite item that you own? Quick fire.

Markus Sauer (00:45:43):
Motorbike.

Peter O'Toole (00:45:45):
I had a feeling you'd say your Triumph. What's your favorite food?

Markus Sauer (00:45:52):
Uh fish.

Peter O'Toole (00:45:55):
What's your least favorite food?

Markus Sauer (00:46:00):
Hmm. There are a few, vegetables I didn't like that much. It's not that I hate vegetables. I eat vegetables. Even

Peter O'Toole (00:46:11):
You sound like a small child. Love your meat. Don't want my vegetables,

Markus Sauer (00:46:14):
No Salad, no salad

Peter O'Toole (00:46:17):
I want to ride my motor bike.

Markus Sauer (00:46:19):
No, no, no. That might give the wrong impression. So I'm from time to time. But there are a few things I don't like that much.

Peter O'Toole (00:46:27):
I am joking. Beer or wine.

Markus Sauer (00:46:29):
Wine

Peter O'Toole (00:46:32):
Oh, okay. Oh, this picture, if it's wine. So

Markus Sauer (00:46:37):
This is wine in my hand. So this is, I mean, I'm not a Bavarian, I'm not a Bavarian, but I'm, I'm at the University of Regensburg and Regensburg is North Bavaria. And they are this festival every year where people wear even students, they wear these typical Bavarian things like lederhosen and Dumble women. Yeah. And we are doing that day. And from time to time with the group to go for an evening, Yurts tent directly translated and try to have some fun, but you see that I'm drinking wine. If you carefully look

Peter O'Toole (00:47:11):
Yay. Yeah.

Markus Sauer (00:47:13):
Or is it Beer.

Peter O'Toole (00:47:14):
It's beer Isn't it all.

Markus Sauer (00:47:16):
Then, then I, then it's

Peter O'Toole (00:47:19):
It looks like a large glass of white wine.

Markus Sauer (00:47:22):
I don't know. But maybe it's no, it looks like white wine to me on the left. It's water. Two girls and mine. I think. I don't know, but I prefer definitely wine. Okay. If I had too much beer in my life,

Peter O'Toole (00:47:38):
Would you rather read a book or watch TV?

Markus Sauer (00:47:42):
Oh, I read non-scientific book. I read maybe once a year TV. I also do not watch TV that often that maybe I turn on TV at 11:00 PM or 1130 and watch the news and maybe just I'm scrabbing through the program. So that's what I'm typically doing sports. Of course. Soccer I like

Peter O'Toole (00:48:10):
So who's your football team. So who do you support?

Markus Sauer (00:48:17):
I don't know. I don't know. No, no,

Peter O'Toole (00:48:22):
No favorite football team.

Markus Sauer (00:48:25):
If, maybe Bayer Munich maybe, but usually I try to be to be objective. Yeah. As long as long as you can. I mean, of course, European soccer tournament or the world championship for the German team. I mean the last two, few years it was a disaster. So

Peter O'Toole (00:48:50):
As a Brit, we know what that feels like for many

Markus Sauer (00:48:58):
This wasn't pity, this wasn't pity. I'm so sorry for that. It was so deserved. It was so deserved that you won against Germany. It was the most deserved win I've ever seen yet.

Peter O'Toole (00:49:07):
Yeah, but I've got to say Italy were very good winners of the Euro 20.

Markus Sauer (00:49:13):
Okay. They were also good. Let's say that they were also good by the way. We are still, I'm still playing soccer. I once a week we're playing soccer at 8:00 AM in the morning from 8:00 AM to 9:00 AM with my students.

Peter O'Toole (00:49:25):
I don't have a picture that

Markus Sauer (00:49:29):
No, sorry.

Peter O'Toole (00:49:32):
I don't have a picture of you playing football

Markus Sauer (00:49:34):
We usually don't take pictures when we play soccer.

Peter O'Toole (00:49:37):
Yeah. Okay. I guess in today's selfie world, I'm surprised it's already there where that's right, right. Okay. So next Elmi we'll have you on the team at Elmi. Okay. The Elmi is always a football game pre Congress. So I mean the commercial companies. It's good. Fun. It's good. Fun. What about music? What do you like listening to

Markus Sauer (00:50:06):
Mixed sometimes classical music, time to time classical music, but usually I hear radio, the radio station though. Okay. Typical pop songs. You're

Peter O'Toole (00:50:20):
Okay. So popular music.

Markus Sauer (00:50:22):
I was with hard rock, like status quo and, and, and whatsoever ZZ top. And maybe if you, if you, if this tells you something ACDC as well, but today,

Peter O'Toole (00:50:36):
And you said status quo,

Markus Sauer (00:50:38):
Status quo as well. But in the early days when I was 15 years old, 14 years old, something, yeah. I guess rates afterwards.

Peter O'Toole (00:50:46):
I think this is your grandchild.

New Speaker (00:50:49):
That's my grandchild Is exactly in my office. Jolie so she's now 11 months old and she's really the pride and joy I have, even though I was, I was, I was not angry, but I was, it was kind of strange feelings. You hear that you're going to get grandpa now and you don't feel like a grandpa. So in the first moment it was not a shock, but it was like, oh, how will I handle this situation? If somebody is calling me grandpa. But finally, I mean, it's real, real, super happy with her

Peter O'Toole (00:51:26):
And I'm sure she'll be super proud of you except for confessing. You like status quo.

Markus Sauer (00:51:32):
I mean, this was when I was 14. I will never sport. So later on it was deep purple rather.

Peter O'Toole (00:51:37):
I wish you'd never, so I've got the song, I've got some of the status quo songs going in my head now, Are you a night owl or early bird.

Markus Sauer (00:51:50):
Ahh depending, but I would say rather early bird.

Peter O'Toole (00:51:54):
Okay. And who who's been your inspirations inside and outside of work

Markus Sauer (00:52:03):
Inside of work my inspiration is discussed discussion with a few people who are, I mean, I'm a trained chemist. Yeah. I love to, to discuss my opinion of things then with biologists on the one hand side and with physicists, what I call hardcore physicists on the other side, because this is quite inspiring me because they have a completely different view than I have for many things. Yeah. So this is in work and at best case scenario, we do that with a bottle of wine outside family, friends sports I mean the best ideas come when you go jogging after 20, 30 minutes.

Peter O'Toole (00:52:53):
Okay. We talked, I guess we talked about some of the most challenging times through before the Nobel prize was awarded and the difficulty with peers at conferences and the fights. So I'm presuming that's one of the most challenging times you've had. But what about the most fun time in work? What parts of your career, if you could relive a moment in your career, which moment would it be?

Markus Sauer (00:53:17):
It would be the moment when we started to, to try to detect single molecule for instance. And maybe I, I likely remember a time where we joined forces three post-docs. We won three German postdocs Claus Seidel, maybe, you know, Claus Seidel from Dusseldorf now. Okay. No worries. Claus Seidel, kristoffersen. And we met, it was 96 that time, 95, 96. We met always in Göttingen at the Max Planck Institute for two or three weeks. And then we work day and night, day and night, I hadn't never lost for the tires Claus in Gooding and had a Thai surf laser. The other guy had a oil immersion objective and was an expert in alignment of lasers. So we always, we joined forces and we worked about 20 hours every day. And we're, it's so much fun even though we detected only the impurities in the oil, but it was so much fun in these nights trying to detect the signals

Peter O'Toole (00:54:23):
Told you you'd miss that at time earlier on I said, but you miss being back in the lab. And that moment, that, that period is one of that, that innovation, the uncertainty of it. We, we all come up to an hour, mark. I think I have a question for you. Where do you, where do you see the future going? What do you think is the next big thing that has to be solved? The next big challenge? What's the next technology that needs,

Markus Sauer (00:54:54):
This is from my point of view, but I think what is, what is needed now is a technology which allows you, I mean, in the, in the last two years super-resolution microscopy developed into direction that higher, faster, further pushed the localization precision down to a few nanometers. But I mean, this cannot be translated in special resolution at the end of the day because we do not have the labeling technologies. We do not have the it's not efficient enough that the detection probability is not high enough and things like that. So we will always end up in a diluted signal and therefore never be able to achieve real sub 10 nanometer resolution in a biological sample. So I think we have to find ways to efficiently label sites and proteins with small displacement. So there is this way of unnatural amino acids, for example, to introduce them. But the other thing is to find ways that allow standard labs buying a standard microscope from a company or commercial microscope to achieve three colors, 3d 20, 30 nanometer and biological samples,

Peter O'Toole (00:56:20):
They're getting there slowly.

Markus Sauer (00:56:22):
Yeah. And I, think the way to do that is by combining the technology we already have with expansion microscopy. So this is what I'm, I bet. Oh, I'm sure it will be realized. Sooner or later for everybody,

Peter O'Toole (00:56:38):
But then we want to do it in a live scenario. And actually I was a bit cool. I said, we getting there slowly. I think that's a bit harsh of me actually. I think the pace over all the time, actually, I think microscopy had, has been extremely fast moving over the past 20 or so years. But yeah, we, we still want it live, I think as well. Could we take expansion? We won't tick that box yet, but actually my students actually today is playing with expansion microscopy. So I look forward to seeing How that's going.

Markus Sauer (00:57:13):
You're focusing a lot on expansion microscopy in the moment I'm remembering I have now what is called as synergy grand running from the ERC on expansion microscopy together with Ed Boyden and Silvio Rizzoli from Göttingen in to push expansion microscopy really to the molecular level, we'll see what we can achieve. But but this is of course not live. I mean, live cell, I think it's super fascinating Leica lightsheet methods. They made a real change to me. And we also set up a Leica lightsheet. So we, we, we, we, we use the in principle, we, we built the same system, Erik published in the science paper, but there are also now companies on the market. They sell quite reliable, lightsheet microscopes and

Peter O'Toole (00:58:09):
On the invert as well. So again, I overcome some of those problems with that executive

Markus Sauer (00:58:14):
Samples, sample preparation, because this is library, this is the pain in the neck to do, to place all the samples on these tiny cover slips and then move them somewhere in. And this is well, but this is overcome. Now, this has been overcome by this inverted Leica light sheet microscopes. I did measurements on it. I don't have one in my lab, but I did measurements on it. And I have to say I was quite impressed by the speed. But then the next problem comes with data handling, I mean this sheer size of these terabytes, but

Peter O'Toole (00:58:47):
We share those problems with the structure IOM, microscopists, and yeah. Yeah, for sure. Did you know, you make fields to metabolize the mass spec field? Its problems are short term because computing is always used to go faster than us. So I think those file size size of the file would have been very problematic as they were for confocal back in the early nineties. Yeah, the files got so much bigger of its surface storage thankfully. Marcus. We are on the hour. So I'm going to say thank you very much for joining us. I've got to say it's been a real pleasure. You actually, a lot of previous guests, you chimed really nicely with a lot of things that are being said from a lot of the different guests in the previous podcast, which has been really nice. So thank you once more for agreeing and joining and I love the pictures. Thank you everyone for watching and listening, please. Don't forget to subscribe to the channel and go back and go, go and listen to Petra Schwille herself. Talk about it, to listen to Eric Betzig, talk about lattice lightsheet. There's so many people that were very similar, but different aspects, different angles and different viewpoints. Marcus, thank you very much.

Markus Sauer (00:59:59):
Thank you please. It was my pleasure. I really enjoyed it, especially the atmosphere. Thank you so much.

Peter O'Toole (01:00:08):
If you enjoyed today's episode of The Microscopy, check out some of the previous ones featuring. So Markus's colleagues and friends, including Petra Schwille, who he skis with who discuss his work life balance

Petra Schwille (01:00:23):
You get a bit addicted to having kids. Once you have the first then,

Peter O'Toole (01:00:29):
And the episode with Nobel Laureate, Eric Betzig, who covers the challenge is in his career.

Eric Betzig (01:00:35):
I was an abject failure up to about age 45. Okay. So why should I waste a lot of faith? You know, when I was 45, I think there's an alternative universe close to a year in which I'm an unemployed, divorced mechanical engineer living in Michigan.

Peter O'Toole (01:00:51):
While, Richard Henderson discusses the perils of the Scottish winter.

Richard Henderson (01:00:57):
Then it turns out riding a motorbike or a scooter in Scotland in the winter. It's not very clever is a dangerous thing actually.

Peter O'Toole (01:01:07):
And listen to Elizabeth Hillman chat about her favorite publication.

Elizabeth Hillman (01:01:12):
It was like totally out there. And it was a real struggle as well to get that published and then a real sort of delight to have a lot of people come and be like, oh my gosh, you know, you, how, where, where did you come from and how did you discover that yes.

Peter O'Toole (01:01:26):
Visit bitesizebio.com/themicroscopists to watch or listen to all these episodes. Really some truly inspiring stories, facts to learn about. Thank you.