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Intro/Outro (00:00:00):
Welcome to Flow Stars candid conversations between Dr. Peter O'Toole and the big hitters of Flow Cytometry brought to you by Beckman Coulter at Bitesize Bio.

Peter O'Toole (00:00:11):
Today on Flow Stars. I'm joined by John Tigges Principal Associate in Medicine at Harvard Medical School. And we chat about his passion for training the next generation of scientists.

John Tigges (00:00:21):
We try to give them things to help their career, not just, Hey, come in. And you're an extra set of hands.

Peter O'Toole (00:00:27):
Why he was actually really great on this episode because of his hobbies, being a DJ and the MC

John Tigges (00:00:33):
Also, it may be very comfortable on a microphone or in setting such as this where it's not, oh boy, you know, who's gonna see this what's going on.

Peter O'Toole (00:00:42):
The importance of sample preparation in Flow cytometry.

John Tigges (00:00:46):
This is the ultimate of how everybody puts it out there. The garbage in equals garbage out. And if the prep is not done in, in a certain way, there there's, there's no chance.

Peter O'Toole (00:00:58):
And his recent promotion,

John Tigges (00:01:00):
I was putting together my Harvard CV recently for a promotion. Um, oh, it's a good time. I will say it. I got it. I'm here. You know, I'm a principal associate in medicine for Harvard medical school now

Peter O'Toole (00:01:13):
All in this episode of Flow Stars. Hi, I'm Peter O'Toole from the University of York and today on Flow Stars I'm joined by the one and only John Tigges, John. Hello. How are you?

John Tigges (00:01:31):
How are you Peter? And thank you for the, uh, overwhelming intro.

(00:01:37):
You know why that is don't you John, because I believe that you are also a DJ and MC, so I thought beyond just your Flow Cytometry, I had to introduce you in no other way than to give you the proper MC introduction. So we will get to Flow Cytometry, John, how did you get into being a DJ and MC though? First of all, we'll start on a personal level.

(00:02:00):
Okay. So that's a, that's maybe not as interesting a story as someone would like it to be. Um, I needed money so while going to, to school, um, you know, one of the gentlemen that I met, uh, he was a friend that I had met through high school, but then we kind of reconnected. Um, as I like to say, I went to, uh, I went to college and he visited, um, and his dad owned the company. So I sat there and he's like, I need help. And I'm like, alright. And I need money. So it was a, it was a perfect match cuz you know, and then it started off with kind of just doing like the, the pub kind of things and helping him there and stuff. And then ever since it's been mostly functions and weddings, uh, and uh, I found that interestingly enough, it was a great opportunity because not only was it fun, I love music. Um, also it made me very comfortable on a microphone or in settings, such as this where it's not, oh boy, you know, who's gonna see this what's going on. It was kind of just a, here it is. Here's a microphone in your hand and you've got control. So power trip

Peter O'Toole (00:03:18):
Right. I would just say we will get to the science in a moment, but I have to ask as a DJ, what is your banker? The track that you'll put on to try and get people onto the dance floor? Cause weddings are not necessarily the easiest gigs.

John Tigges (00:03:32):
Oh, they are not. And they're hard to figure out because you have such a diversity of age, but, um, we, it tends to be surprisingly some like if you play some Bruno Mars and to all feel because it it's newer, but it kind of has that, that Discoe vibe to it. So usually a good start there or, you know, you get a list of some things and you pick out the one that's kind of not gonna be over the top, but really gets a good kind of feeling in people. And um, a lot of times too, they're just anxious because they've been sitting at a table and you know, had dinner and now they're just waiting. So

Peter O'Toole (00:04:14):
Yeah. So I've been to conferences with you, John. I've been to the evenings where it ends up with a DJ and a dance floor. In fact, I remember Leipzig vividly well and Alfonso who we know other episode has got some good pictures of that, but I remember you did not get up and dance that night.

John Tigges (00:04:35):
Uh, yes. Um, well, I, I guess I, I, I tend to try to be a little more refined, uh, amongst colleagues than when I'm, uh, I'm out and about regularly. Um, cuz even just doing weddings and stuff, I find myself at times, uh, you know, dancing behind my, my sacred place there. Uh, and so , I guess this half is all Irish. So,

Peter O'Toole (00:06:11):
So if you go go going back, if you're half Irish, where were you? Are, are, were you born in the US?

John Tigges (00:06:19):
I was born in the US.

Peter O'Toole (00:06:20):
Yeah. So where about,

John Tigges (00:06:22):
Uh, Rhode Island? So we are east coast, um, still live there, um, because it is a, uh, slightly more fiscal answer to the, the Boston area. Um, you know, working in academia and trying to live in Boston is difficult. Uh, it's expensive. It's, it's on par with, you know, the New York, the London's and San Francisco and stuff. So, um, kind of born and raised there. And that's why the accent is this kind of half Massachusetts, half New York, um, or sandwich somewhere in between there. So,

Peter O'Toole (00:07:02):
So, so I know, so you are, uh, the Beth Israel Deaconess Medical Center, which is part of Harvard, is that correct?

John Tigges (00:07:09):
Correct. We are a Harvard teaching hospital look at that, right.

Peter O'Toole (00:07:13):
so how did you get into Flow Cytometry?

John Tigges (00:07:16):
Uh, so out of, out of University, I worked for an immunology lab and so at, uh, it was actually it's called Roger Williams University, uh, Hospital and it's, um, part of Brown University. So I got a position there and then they said here's a Flow Cytometer. It was a FACS scan. Um, and you know, I, anything with electronics and stuff, I guess maybe that's the whole DJ part, the electronics and things were interesting to me. So I said, okay. And I said, do you wanna try? Sure. And I, whether whether it was a fortuitous thing or not, I was really good at it. So had no idea what I was doing, cuz I wasn't taught, I just sat down and started hitting buttons And became very kind of, uh, adept at at it. And then next thing I know they purchased a, what they call now a legacy Moflo, um, and gave me that

Peter O'Toole (00:08:24):
Ah, the Legacy MoFlo,

John Tigges (00:08:28):
Ah, it was like an erector set. Right. Uh, you could put anything you wanted on that breadboard and, and just go. So

Peter O'Toole (00:08:36):
I, I, I was gonna ask later on what is your favorite instrument you've ever had

John Tigges (00:08:41):
Ever had? Um, I've gotta say in between it was my Moflo XDP because of the things that we tried on it. Um, I mean, this is stuff I guess, that that would interest you from your microscopy standpoint. We, we actually placed a chameleon laser on that, uh, case

Peter O'Toole (00:09:01):
As for the multi photon. So for far red ex citational or to, to excite UV dyes, but using the far red for those who are not with flow cytometry and coherent.

John Tigges (00:09:09):
Yeah. So, um, so we had that in a, from a coherent and coherent also gave me a laser stack. So we had a, we pig tailed in 5 61, 5 32, uh, four 50. We were playing with a bunch of weird laser lines, uh, 8 0 8 0 8, I think there was. And then we had that 10 80 and the 10 80. I loved because it wasn't a simple alignment. See it . So for those who never played with a chameleon laser or, or out in the IR range, you just can't see them. So it became very interesting on how we tried to align that we used a, um, a Nikon camera because your little digital camera is when you, when you take photos, do it in an IR. So they can actually pick up the laser beam. So we were aligning through all the focal points and stuff, using the camera and kept taking pictures. So it was a very, uh, long process, but it was all because we were doing, um, opto injection. So it was, uh, trying to kind of get the red blood cells to open insert stuff inside and then close 'em up. So it it's because red blood cells tend to have this ability to, uh, form after they've been deformed. So that's what we were trying to do. And then we later found out that we were successful. We were able to actually get Dye in. It was, uh, very bright Lucifer yellow, what a wonderful died. Yeah. Um, and so we were able to get that into the red blood cells, but we found out, we read an article that then said that opto injection with a, uh, a 10 80 laser had a 2% success rate . And that if we had just used a violet, we would have a 75% rate. So we were able to, uh, actually do something. Uh, and I, I joke with my colleague, uh, I said, yeah, I said, we were, uh, I guess we can call ourselves the one percenters because we at least did it once. Um, but boy were we barking up the wrong tree?

Peter O'Toole (00:11:31):
That's uh, I love the concept though. I got my background is red blood cell biochemistry. So I quite like the concept of actually punching a hole in it and watching it resealed, but

John Tigges (00:11:41):
Things with deformability through channels. So you get them to do this up and down channels and watch. And the more rigid they are, obviously during disease states, they won't go through and they get stuck in stuff. So deformability, um, their vibrations, he believes their vibrations. Actually there is them talking to one another. And so what he did is he sent it to somebody to put it to music so that each of the different vibrations were actually musical notes. So he was trying to get it that say when they got more rigid in like a sickle cell, if they could play the, uh, dark Vader entrance theme so, yeah, so this is what we do scientifically. We may not, um, we're, it it's slightly in unorthodox. Right. And, and I may have found my, uh, my, my kindred spirit in this world because neither one of us thinks in, in a very, um, straight line, everything is the crazier. It is the more likely we are to try it. So

Peter O'Toole (00:12:47):
Isn't that way you get the eureka moments and the big breakthroughs.

John Tigges (00:12:51):
Yes. This is definitely where they happen. Um, and if nothing else, it's a lot of fun. Um, you know, I'm not gonna Flow Cytometry is what we do, but staring at dots all the time is not necessarily, uh, you know, very stimulating over a long period of time. So within a 26 year career, um, I need something to keep the old mind and the wheels turning. So, uh, you know, just that earlier. You know, I forget the, the little details because we're always looking at the big picture. We, I said, we need to put it on the wall, a little sign that says, you know, two things, it's a go or no, go in lacutanesca. Uh, just throw everything at it and see kind of, what's great. You know, the egg roll principle.

Peter O'Toole (00:13:44):
I, I was just gonna ask, when did, when, when, when, when did you use your first Flow Cytometer

John Tigges (00:13:50):
In, uh, let's see. It would be back in 1996,

Peter O'Toole (00:13:55):
1996. And you've been practicing it for 20 years

John Tigges (00:14:03):
Now. 20 it'll be 26, uh, in mid May.

Peter O'Toole (00:14:07):
I'm I'm just trying to strategically work out your age without actually asking straight out. How old are you? But I guess if you're about 47 anyway,

John Tigges (00:14:18):
Yep. I started flow when I, um, when I was just about to turn 21 and, uh, I can honestly say that I've grown up in The world of Cytometry

Peter O'Toole (00:14:30):
So I, I, we are not so dissimilar in age and I, I do know that I I've gone perfectly going perfectly gray you on the other hand, we've got a great reason for why you've not gone gray you've actually just lost hair on the top of your head. Well, why is that? What is your excuse?

John Tigges (00:14:50):
Um, I'd like to say, you know, the, the classic, uh, job stress and stuff like that. But I, I also believe that the, the two young women I have in my life, um, over the years, they, they have challenged me and, and, and stressed me to some extent where the gray shows up here. Uh, and on, on your chin, all this went here, but yes, uh, doing this is my go to when I'm thinking.

Peter O'Toole (00:15:17):
So just rubbing the top of your head.

John Tigges (00:15:19):
Yeah. So at some point I'm pretty sure that I've just rubbed most of the hair out. Um, so the point where it just gave up on me

Peter O'Toole (00:15:27):
And you're blaming your daughters for that.

John Tigges (00:15:30):
Some of it, yes. I, I guess I , they won't won't watch this, so

Peter O'Toole (00:15:34):
No, I'm now gonna make sure they watch or listen to

John Tigges (00:15:37):
This. I'm not, you know, unless you, you, you do some sort of TikTok dance or you're on Instagram, Peter. I don't know if they'll ever see it. So I

Peter O'Toole (00:15:46):
Do, you know, I forgot. What was your favorite dance move on the dance floor? Here's your TikTok moment, John?

John Tigges (00:15:51):
It could be my TikTok, you know,

Peter O'Toole (00:15:53):
,

John Tigges (00:15:53):
Uh, you know, it's,

Peter O'Toole (00:15:56):
How old are your daughters?

John Tigges (00:15:57):
Uh, they are 17 and 15. Well, I have one off to university next year and the other one starting, uh, her sophomore year of high school.

Peter O'Toole (00:16:07):
So what, okay, so your eldest, what are they off to uni to study? Hopefully

John Tigges (00:16:12):
Psychology and criminal justice.

Peter O'Toole (00:16:14):
Ooh.

John Tigges (00:16:15):
So she is, um, she is off to it's Roger Williams University and they are law school. So she wants to, uh, practice law.

Peter O'Toole (00:16:25):
Well, there's a wise lady to start with cuz there's bound to be more pay in that than there is in

John Tigges (00:16:33):
And plus yes. Uh, nobody, I don't think my younger one talks about medicine. So as I work for Harvard she's, uh, she keeps asking me if I can get her into the medical school. And I said, um, said, very good grades. And um, yeah, about all dad's influence has none, as far as Harvard medical school goes, I said, you know, they let me work here, but that even that they consider to be an honor and privilege

Peter O'Toole (00:17:05):
Actually, do you know what she may want to go to medical school? I would say I, I certainly in Microscopists. There's quite a few medics who left medicine and come back into, uh, into science research.

John Tigges (00:17:16):
Yeah,

Peter O'Toole (00:17:17):
No, I, I would say medics only diagnose and prescribe. They are actually useless without the technologies such as Flow Cytometry to analyze and tell you what the results are and actually useless without the scientists developing them, the, the tools to do that and the medicines to cure that. So arguably she just needs to go to, to some sort of life science or technology research if she wants to make a profound different impact.

John Tigges (00:17:46):
Well, so I tell her these little stories and it's funny you say that because just the other day I was talking with somebody and I was analyzing the results and I said, we really should perfect this and really go through some more. Um, and so he said, well, my boss, another MD wants to get this and he needs the results. So he wants to do this. And I said, okay. And he said, it's clinical. So the bar's really low. I just was like, I didn't say anything, he left. And my two technicians looked and went, so the clinic is very low on the bar. I go, yeah, I, Hey, I said, he's a lovely man, but maybe not who I'm going to if I have a medical issue after talking with him. So it's very, it's funny you say that they, you know, diagnosed and prescribe and that's about it to sit there and go, well, the science we're setting the bar low. Um, so, okay. Um, I'm like, well, I don't know how that makes me feel and should I feel more important or less? I'm not sure. So, uh, it's an interesting concept.

Peter O'Toole (00:18:59):
Obviously we are joking about the quality of MDs here.

John Tigges (00:19:03):
Of course

Peter O'Toole (00:19:04):
This goes to diagnose so many different things, but I do think, yeah, and I I'm sure they also appreciate without we are all part of the same family.

John Tigges (00:19:12):
Okay. All seriousness. We have some lovely physicians that I work with and you know, they're doing preclinical work and just being a part of that is the stuff that makes the job worthwhile, you know, to that when someone asks you what you do and you can actually say, well, I'm working with someone to diagnose, you know, to look at ways to diagnose and, or, you know, treatments for this type disease, you know, whether it's a cancer modality or, you know, like we mentioned anemia, they do stuff with malarias and, uh, all kinds of wonderful stuff. And so it's, it's great to be part of that and be able to put the little Harvard logo next to your name. So,

Peter O'Toole (00:19:56):
Yeah, I'm proud of York. I, but, but yes, I, I, I totally get it. Yes. Uh, I, I can't remember where I was going by next week. We just asked about your daughters and where they're going. Yes. As a child, what did you want to be

John Tigges (00:20:14):
As a child? I, I actually wanted to be an architect. Um, as odd as that seems as a child, I, I was always, and, and this is why I love going to Europe is because there's just that, that different architectural aspect where, you know, the US is relatively young. Um, and so the architecture is, is more modernized. And I, I just, I, I loved when we, I went to Budapest and just the, the architecture there and stuff. So that was where I was going. And my first year of, of schooling, uh, you know, in, in college is what I went for architectural engineering. Um, and again, it was one of those situations where I was taking biology and chemistry classes as part of my, my undergrad requirements and got talked into, um, changing majors by my, uh, my biology professor said, you're incredible at this stuff. And I said, that's great. I don't really care . And so, and it was my third year of differential equations that I decided and architectural engineering may not be, what I was born to do. Um, because I, I I'm like this. All this math is really hurting my head. Um, and so I, I discussed it with, uh, those professors again, and changed my major.

Peter O'Toole (00:21:41):
So differential equation sums up why you didn't want to be an architect, nice play on words there

John Tigges (00:21:48):
It was. So, uh, I, I differe equated to a different position in life. And it was funny because it was a professor who was a triple PhD teaching that particular class. And this gentleman was way above my, um, my mind capabilities and, uh, I wizard or of some sort. Um, and it was interesting because when I went back home and said I was changing majors and, uh, discussed with my father, my, my issues with a, the differential equations in this particular professor, my father said, you've gotta be kidding still there. So my father actually had him at a different University and struggled immensely and changed his major. So this particular professor was, uh, very well adept at, at, at destroying the, uh, the entire Tigges's family. So,

Peter O'Toole (00:22:49):
Yeah. Oh, wow. So, so, you know, one, you are answering my questions before I get to my questions. And one of those is, you know, who has been one of the inspirations or most influential on your career to date,

John Tigges (00:23:01):
Um, career wise, influential? Uh, my, you know, obviously I, I do credit family. Um, my father taught me a lot of good life lessons, if not career lessons, per se, um, you know, his little anecdotes I still use to this day. And so, um, you know, and then just actually the, the first and still kind of one of the, the only in the family to graduate from a University. So that was a huge accomplishment. And I was, I was pushed by my dad was very much a work oriented education oriented individual. Um, even though he did not graduate, he was very successful in his career. He, uh, he was a, he worked for a telephone company for 35 years, um, and moved his way up through the ranks. And then once I got into Flow Cytometry in general, it was a gentleman by the name of, uh, Nicola Kouttab, a Cypriot Um, so my first experience with the, the Greek culture per se, and, uh, and he was very influential because he, I think he taught me more how to be a good boss than anything else, because he was very, as long as the work got done, he was okay with anything else. It was like, he told us from beginning, usually when you get someone starting a position, you don't start off by telling them, Hey, you wanna take a two hour lunch, go for it. You know, you need to run errands, go for it. And I was like, oh, okay. You know, being 20, uh, 21 at the time. And, and someone telling you, you know, live your life. It was like, wait a second. What, but he was just, Hey, you know, if you wanna come in at five in the morning and get everything done and then take off at, at, at noon. And so I, I've kind of taken a lot of that teaching into how I try to, you know, manage the, the, the core facility in, in the people that I had, the, the privilege of mentoring and work with me.

Peter O'Toole (00:25:13):
So you had, uh, for many years, a partner in crime

John Tigges (00:25:18):
Ah,

Peter O'Toole (00:25:19):
Yes. Vasilis. Yes.

John Tigges (00:25:22):
It's my work husband. Um, so, uh, my first, the first place that we were at, the Roger Williams that I mentioned, um, I met him there, um, showed up kind of on my doorstep. Uh, I joke with him and say, you were like the, uh, the, the baby that was left at the orphanage orphanage cuz you just showed up one day and no one knew that he was coming to work with us. Um, it's a, it's an interesting story of, um, you know, he had a relative here that, um, had an illness. So his mother, he was supposed to go back to Greece. He was a podiatrist and his mother asked him to stay and, and, you know, stay with her and make sure everything was okay. And his, his aunt, that particular aunt was very friendly with the president of the hospital. So the president of the hospital gave him a job that even my manager did not know was a position . So, and then, um, we worked together, I moved to Beth Israel here and, um, he was in between what he was going to be doing next, uh, was talking with my wife, uh, via Facebook. And I said, I'm looking for somebody. So we came back and so total, I think we worked together over like 18 years And it was weird because when he finally, um, moved on to industry and doing some clinical, uh, work for Beckman Coulter, there was many a person such as yourself that, uh, felt, felt and said, it was like, what, what could be sacred anymore if we broke up so, you know, the, the worries of, oh my, you know, work relationships, you guys were the, were the ones that everybody else felt, you know? Um, you know, now it's, it it's like Peter and Karen. Right. It's I can't imagine ever hearing a time of, of, of Peter without Karen or Karen without Peter. That, that was, that was how we were, um, obviously you were still friends, very close. Um, he's my younger daughter, Kylie he's godfather. So, you know, we we're like we're family, so that's not gonna change.

Peter O'Toole (00:27:36):
No. Um, say hello to him

John Tigges (00:27:39):
When you've been. Oh, definitely. Uh, his daughter, his daughter was at my house yesterday, so yeah, hanging with my wife.

Peter O'Toole (00:27:46):
That's very cool. Uh, I'm gonna change that question. Uh, I'll come back to that one in a minute. Nano cytometry. So actually, I, I, I look, I don't do a lot of background research. I shouldn't need to do background research on you, John. I know you fairly well, but actually in your description is one of your expert areas of development of expertise is nano cytometry. What on earth is nano cytometry.

(00:28:11):
I'd like to joke and say it's really small cytometers um,

John Tigges (00:28:15):
I, I just thought through sending off Mork and Mindi. Yeah.

(00:28:19):
Nano. Yes. Ah, yes. Uh, Robin Williams is one of my favorites, so great reference. Ah, wow. I don't even know where you possibly pull that's that's coming from the, uh, the, the stratosphere there um, so no, in all seriousness, um, we got into nano cytometry about eight years ago. Um, someone approached us and we had no idea. So this is looking at things that are below the one micron in size. Um, they can be known as extracellular vesicles. They have names, you know, the, the whole nomenclature was debated for years and still is, um, what should they be called? So, um, and it's just being able to look at these things. And let me just tell you that there are many a time that I want to go back to just plain old cell biology. Uh, cuz when you start looking that low, as you probably know, when you decrease a threshold that much, you see everything and your job becomes getting rid of all the extraneous materials and that can be simply you find out that people have contaminated, uh, you know, HBSS or any, uh, you know, PBS, I found out our Millipore water is pretty dirty considering, you know, everything else. And, and so it's, I think it's a wonderful, wonderful science. It has brought, um, a lot of recognition to my core has given me lots of speaking engagements. Um, I, I would also say that most of us probably don't know 80% of what we should in order to do this correctly. Um, I mean standardization and flow cytometry itself is problematic that whole rigor and reproducibility, it is a nightmare to try to do this with, you know, instruments that were not built for this. We're basically, we're trying to use a hammer to do everything, you know, to put in the screws.

Peter O'Toole (00:30:29):
You say instruments aren't built for it. Light

John Tigges (00:30:33):
Is not built for it.

Peter O'Toole (00:30:35):
Isn't built for, I became

John Tigges (00:30:40):
Extremely, extremely, uh, doing darkfield microscopy. Um, so darkfield is one of those things that actually kind of works with these guys. Um, when everything else is dark, you can see the little guys that are the only light. Um, and so it it's, you know, for, for those who are not familiar with darkfield I always think about it like starry night, right? When everything there's no clouds, the sun is the moon is completely blocked and you've just got those black nights where you can see the stars perfectly. And there's the big dipper in O' Ryan. Um, it works the same type way and that my microscopy guru, um, that's how he always describes it to us. He's like, it's like the starry night and he's um, it's interesting. I I'll ask you this. Isn't supposed to be about you, but are you also, uh, big into astronomy? A lot of the microscopists, I, I know are love astronomy and the telescopes and

Peter O'Toole (00:31:41):
Not, not massively into astronomy. Uh, certainly I know my consolation and I'll do a bit of star gazing occasionally. Uh, Chris bizarrely, Chris Lintott is a, we have a program called Sky at Night in the UK. Uh, and the presenter of that is Chris Lintott. Who's actually also a guest of a podcast series called The Microscopists with us. And this is, this is the crossover between the astronomy and the scientists and he to actually understand the universe better. He, he started so called Zooniverse where you, you use citizen science to start helping analyze all this data, you know, minds, loads of data and you just a computer can't do you need to train the computer? So you need the human input. So he is got thousands, probably hundreds of thousands of volunteers doing this, but of course microscopy needs to save help. And so then Zooniverse now has a for essentially called Etch a cell with, uh, Lucy Collinson, uh, Chris Peddie, uh, oh gosh, yeah, mark down in, uh, London that do just this and, and they, they, the loads of volunteers out there helping solve counter by just literally drawing round images, cuz if computer cannot do it as accurately as a human. So, so, so no, not as astronomy a little bit. Uh, but it certainly has those relations through and uh, maybe bird watching is more my thing. So I still have the telescope that I can use for both.

John Tigges (00:33:12):
Okay. Cause yeah, cuz one of our big things eventually is to take a pilgrimage to one of these, you know, super telescopes, uh, whether in Africa or I think there the there's one in Peru. Um, so the two of us are like one day, this is where we need to go. So it it's interesting how all these things wrap in like you said, tie into one another and how just the interest in, in optics itself spans so much and just, you know, and how it, it gets the, the juices flowing. I mean, my table's usually cluttered. They cleaned it behind me for today. Um, it's usually cluttered with, with photonics magazines and spectra and all these, uh, these, these little fun things that I think when I'm on the train reading, the, the individuals are, are like, what is going on here. Um, because it's just, I don't know. I guess I certain things nerd me out. Um, I, I realized that this passion when, um, Cyto was in Glasgow, no Edinburgh, but I went over to Glasgow where a coherent factory is. Yep. And when I actually came back from it, I called the family and was talking to my wife and my two daughters and was like, oh my God, I went to the laser manufacturing facility for coherent and they had all these lasers there and they were talking about, you know, how they can manipulate, you know, the neurons and all this fun stuff within the mice and the brain and, and how these technologies can really, you know, with Parkinson's and all this. And I was just going off for like about a 10 minute monologue. And at the end they went, huh. And I'm like, oh man, I'm like, I just kind of nerded out. I said, I gotta realize this isn't your Disneyland. This is my Disneyland. And they're like, yeah, cuz I was like, you need to come and you need to see this stuff. And they're like, doesn't sound like a very fun vacation

Peter O'Toole (00:35:21):
Now, but a very much a busman's holiday. I, I think if you call it in the UK where you go off and do a similar job, but on your holiday, when you go overseas, I liked your analogy to astronomy actually, because even going back to nano cytometry, you know, I, I use the, the analogy of the stars. I, I I've signal to noise. You know, you can see very few stars when there's lots of light being scattered in the sky. In fact, you can't see a single star in daylight just cuz it's the light scattering, but your fluorescence it's still shining going out. Or in the astronomist case, the star is still shining. The same amount of light it's not changed. You just cannot see it over that background. But for nano cytometry, it it's all the other particles that are getting in the way of those really faint stars that are interfering with the image. And this is what scares me about nano cytometry. This is where, this is what I hate. The fact we can get so small and for bacteria and some EVs, we can see them, but there's always going to be particles. It it's, it's not black and white. It's not, they're all of this size. There's a slope into the noise and we can't see them all. And how do, how do I really struggle to cope with knowing I can't see everything on my cytometer or microscope whichever way it is.

John Tigges (00:36:39):
Yeah. I think that is the most frustrating part. And I'm sure that seriously has contributed to some of that hair loss we spoke of, um, is just trying to figure out, I always, always trying to figure out how can I really get that signal to noise to be better? How can I eliminate more dark noise? And this is the ultimate of how everybody puts it out there. The garbage in equals garbage out. And if the prep is not done in, in a certain way, there there's, there's no chance. I mean, there's some kind of, okay, let me see if I can save your sample when you're talking about cellular biology. There's nothing. I mean, if you are not, I like to tell people, I said, you you're trying to get donors, right. But you're you are really worried about, Hey man, what did you eat today? Have you had a latte? Your, your lipoproteins like your APO lipoproteins that you can see cloudy layer that just sits there. And then once you get rid of that, you're talking about Kylo microns, which span the entire size range of extracellular vesicles. And again, more lipoproteins and you can generate my cellies. And so it's just horrible kind of thing. And, and companies are, are on this kind of arms race of who can put out these products. And so size exclusion chromatography has become this kind of go to thing where centrification was ultra centrification was a huge thing. Right. And then people came about and said, no, no there's problems there. And then size exclusion became a thing. And it was like, well, there's problems there. And then it was like, well, what about, you know, uh, flow fractionation and, and stuff. Uh, well there's some problem. So now you read papers that are like, okay, we did three processes. Yeah. And like you said, now I can kind of see things and I know clean, but I lost 80% of anything that could possibly be useful. And so every, every single, I don't know how many of these nano talks that you may have attended or on a webinar, almost everyone in some slide has the iceberg. Um, and the, you know, we can only see the tip of the iceberg. And so I like to think that we're getting a little bit lower and we're starting to see things that, that are more interesting. Um, you know, and I like to think that some of our work has been instrumental to that. Um, why not? You know, why can't, I think that, um, doing science,

Peter O'Toole (00:39:18):
Ah, I've just had a brainwave, but do you know what I'll, I'll talk to you when we finish that, John, on that one, i'll keep that one till afterwards. I've just had a thought on that. I think what what's really, yeah. Nano cytometry and it's not just nano cytometry, that struggles, it shows that technology has evolved really rapidly over the last 20 years. And there's still such a long way to go. There's so much still to develop instrumentation wise, probe wise and to understand so research wise and actually anyone listening, who's, uh, a new sci scientist coming in and wondering where careers are. Wow. There's so much still to be learned and developed in this area. I, I think 10 years ago I was thinking cytometry may burnout. Other technologies were coming to the fore and oh my goodness, it didn't burn out. It kind of just had little plateau and then it's rocketed again. And if you look back in time, it, it took off from the zero and it started with a plateau and then went up then plateau then went up and plateaued and it's up. And the gaps in the market mean there's still plenty of rapid rises still to come. So we are good throughout until our pensions.

John Tigges (00:40:30):
Yeah. I think there's two. When I look at it, I think it's twofold. You, you couldn't, I think one is we're very good sales people. So while flow, like you say, it kind of, there was this kind of just this level is we started getting the immunologist to be like, Hey, without us, you can't figure anything out. And so we need you to push the companies to stop making ways to look at multitude of things simultaneously. So 30, 40 colors, you know, and, and that kind of trend went. And then the flow cytometry said, oh, it's great. You can do Western bloting and some PCR, which even with your little extracellular vesicles, now, if you want a multiplex, you need us. And so we're very, uh, we're very good at adapting selling our, our skill set and, and kind of, um, you know, self-promoting, , you know, self promoting. So, you know, we're, yeah, we're important here. Um, because like you said, there's a lot of things that can be done computationally in a very good mathematician can, can take a lot of the burden off of analysis and what we're trying to do. Uh, and I know I've gotten to be very good friends with a lot of mathematicians. Uh, people that are, are, are Whizzes at computational analysis, um, and that's where things have gone. And then the second portion of that is I, I say to people, we've had a lot of new developments in flow cytometry waiting for that next novel thing. I, I, I kind of caveat that by saying new means that it's just the technology and we're using it for a different purpose, novel, the, the flow kind of world upside down, um, and things that were done between the flow part and just cytometry when it was invented, just looking at air and air, you know, same principles, but there was no fluid to it whatsoever. Um, it was looking at for Aero aerosolization in, you know, things of that, that nature. So I'm like, what would be that next kind of over do we lose the fluids again? Um, you know, the lasers become some sort of other thing. And that's why these conversations with guys like Bill Telford who are laser gurus are so fun, um, cuz of the thought process and even looking at, Hey one really kicked up white laser and lots of pinwheel fun, um, or prisms and you know, that's, I like to say flow is about smoke and mirrors. So

Peter O'Toole (00:43:06):
Yeah. I can't see us putting our electron beam through the flow cytometer just yet, but, but never say never, never say, never

John Tigges (00:43:16):
Say never

Peter O'Toole (00:43:17):
Changing a little bit. What, what has been the most challenging or difficult time you've had in your career to date? I say to date doesn't mean you're gonna get more difficult times coming forward.

John Tigges (00:43:30):
Something I didn't, um, at 11 o'clock, someone will be giving you a little piece of pink paper and a cardboard box to put your stuff in. Um, no, seriously I think the most difficult aspect and I, I, and I think a lot of people have the same thing and we mentioned it is the revolving door. Um, it's great to meet, um, you know, a, a bunch of people, but I'm the kind of person that, you know, I put a lot of stock in relationships and put a lot of stock into, you know, meeting people and, and learning about them and, and understanding their goals and stuff. And so when they leave, it's almost, uh, you know, over the years it's like little pieces of you are taken with them. And so whether it be someone that's worked with me directly within the core, um, you know, I, I was putting together my Harvard CV recently for a promotion. Um, oh, this is a good time. I will say it. I got it. You know, I'm a principal associate in medicine for Harvard medical school now. Congratulations. Yeah. Thank you. Um, and part of that was a list of, of mentees. That was a huge thing they wanted to see and just looking at that list, it kind of, wow. Look at all, all the individuals and it wasn't so much like look at all the individuals that I've had some influence on. It was more look at all the individuals that have been part of my life and now have moved on and, and, you know, that's difficult. That is a very difficult part. Just seeing them all go over time and, and, you know, that's the nature of the beast. I get it. But you know, some have come back, it's been great. Um, you know, seeing them again in different positions and, and being able to interact with them on a different level. Um, but you know, we said, we talk about it later, but you know, individuals that, that you've sent along the way from York, like this gentleman, Andy stone, um, Harry Potter, lookalike contest winner, um, , he loves that one. Um, but he embraces it. He dresses it up like that for Halloween. He said, but individuals like that's

Peter O'Toole (00:45:44):
That's not dressing up is it if you already looks like it? Oh,

John Tigges (00:45:46):
I guess it's not, I guess it's, uh, just, it's just putting on a scarf at some point. Um, but he, he's such a wonderful individual. And I use that as an example that, you know, eventually he will come back to York. He wants to go back to, you know, to England and the UK. And so just the, the six months that I've spent working on nano cytometry with him and, and just getting to know him is unbelievable. I actually have a Christmas card sitting above here of, of Andy of Andy and his, uh, his significant other in his kiddy. So, um, you know, you make relationships with these individuals and, and it just becomes more and more difficult to, to lose them. It's

Peter O'Toole (00:46:33):
And on that, Andy, if you are listening or watching this, where's my Christmas card, I just out of interest.

John Tigges (00:46:38):
Yeah.

Peter O'Toole (00:46:39):
Be back to York at some point. So, so make, get that card sent,

John Tigges (00:46:42):
He said it was in the mail. hold on, hold on. Let me, let me see. He wants to know if he, if you moved, he thinks that may have run the problem.

Peter O'Toole (00:46:53):
Thinking about those. Some of the more challenging times it's sort of losing a bit of you each time they go. I, do you feel the same about, we've talked about this in the past. It's quite interesting to hear your views on this, about your attachment to the facility itself as well.

John Tigges (00:47:07):
Yeah, so I think that's it. We we've started talking about this in the past and I'm sure others have the same. When I came to Beth Israel, we were a, uh, FACS scan and, um, in an FC 500 facility, um, with a, a one employee in, in the, for the Flow core, um, the guy hired and, uh, and you know, hundreds of users for two instruments. And so to this day, you know, we've built, we've moved to a, a new building about 10 years ago. Um, built a, a wonderful facility. We now have, um, it's my, myself and we, we have employees there's one open position still. And, um, I have, uh, three interns all working in the core on projects.

Peter O'Toole (00:47:55):
So John, how many staff did you say you have currently working

John Tigges (00:47:59):
Two,

Peter O'Toole (00:48:02):
Say again, sorry you broke up there.

John Tigges (00:48:03):
Oh, sorry. We have two employees. Yeah. Two technicians, one open position. So there should be three. And then, but there are three interns from a local university.

Peter O'Toole (00:48:13):
So about six staff currently working in the lab,

John Tigges (00:48:17):
Interns though tend to be more, um, they learn the flow cytometry. They, they do, um, work along with the, the researchers, but they have projects that we give them to assist on. Um, and it's great because they, they love it. Um, we have a young lady who's leaving us in may. She's been with us for two years as an intern. Um, she's getting a job in industry, but she, uh, will leave with, um, being first author on two papers and a, uh, co-author on another paper. Wow. So we try to give them things to help their career, not just, Hey, come in. And you're an extra set of hands. Um, and so that's great. But again, even with them, it's like, because of their age and my age now it's like children, right. It's like watching your children. And while, like we said, it hurts and it's a little piece of you that goes, there's also a sense of pride. Um, you know, and, and that feeling that hopefully they, they got something from you and will take that along in their careers. So it's interesting. There's as you said, dynamics and the same thing, if I ever decide to leave this facility for, I don't know what might be greener pastures or someone finally convinces me that the industry is not, not the dark side. Um, and, uh, then I would love to see someone here who, uh, you know, I feel is just as dedicated. Um, because again, the facility itself feels like a child so and brought it from a baby right up into its, its now adulthood. Um, and that they can maintain the, the, the quality and, and even hopefully, as you know, and we've discussed, it's not just about status quo, it's the next person who can push it to a higher level. Um, you know, you always wanna see the people that you mentor go above and beyond. Maybe it's just me, but I always wanna see them better than I am. Um, you know, I have no problem with that. My, I don't have an ego in that sense of, oh my, you know, someone you, I trained is better than me. No, I, I think if they're not, I haven't done my job. Um, and so I see my job as an, an educator and a, a mentor, not just the, the guy who looks at some instruments and they laugh at when I take them apart.

Peter O'Toole (00:50:41):
So I'm gonna, I'm I'm gonna think about this. You've got all this time, some quick fire questions for you.

John Tigges (00:50:49):
Okay.

Peter O'Toole (00:50:51):
Early bird or night owl

John Tigges (00:50:54):
Night Owl

Peter O'Toole (00:50:56):
DJ obvious, neat or messy.

John Tigges (00:50:59):
Organized confusion.

Peter O'Toole (00:51:02):
Organized confusion. Okay. PC or Mac

John Tigges (00:51:06):
Mac.

Peter O'Toole (00:51:07):
Oh, McDonald's or burger king.

John Tigges (00:51:13):
burger king.

Peter O'Toole (00:51:13):
Burger King

John Tigges (00:51:14):
Yes.

Peter O'Toole (00:51:14):
Oh,

John Tigges (00:51:16):
They give out little, uh, paper crowns. .

Peter O'Toole (00:51:19):
Is that not gonna go there? Tea your coffee,

John Tigges (00:51:23):
Coffee.

Peter O'Toole (00:51:23):
Coffee. Wine or beer?

John Tigges (00:51:29):
Beer

Peter O'Toole (00:51:30):
Beer. Okay. Chocolate or cheese. Sorry, give us an herb before it, John, just keep breaking up there. Chocolate or cheese,

John Tigges (00:51:40):
Cheese,

Peter O'Toole (00:51:40):
Cheese. Uh, so sweet or savory. I'm gonna guess savory.

John Tigges (00:51:47):
Savory,

Peter O'Toole (00:51:47):
Definitely. Okay. TV or book. Is that a book?

John Tigges (00:51:55):
Yes, a book. I would. I like,

Peter O'Toole (00:51:58):
So what, what are you reading at the moment? Uh,

John Tigges (00:52:02):
So I'm waiting upon, uh, reading once my, uh, my technician Brandy is done with the Witcher series, so I have watched it, but I have not read the books. Um, so previously for me was any crime novel. Uh, some of them being, uh, so I, I guess whether it's good or not, I have this infatuation with the criminal mind or, uh, you know, your serial killers. Uh, and so sitting there going, I don't know if it's just the, the biology aspects, but I'd love to see kind of brain pulses and scans of, of these individuals. So, and some of the books have it in it. So it's very interesting. It's intriguing.

Peter O'Toole (00:52:50):
So that's where your daughter gets it from, I guess then

John Tigges (00:52:53):
Yes, they have. Um, so my younger daughter is actually, she likes watching, but she's read like the whole Harry Potter series, like three times. Um, and she actually, from last time I was in Dublin and I had the kids with me. Um, she actually bought a book on the, um, the famous Irish criminals and from like Kilmainham Gaol. So, which was very intriguing to me. So, and it was, she bought it with money that she won off of an individual by the name Tedar

Peter O'Toole (00:53:25):
. Okay.

John Tigges (00:53:29):
Winning money. Tedar in a pub was not difficult.

Peter O'Toole (00:53:31):
especially now that yet. No, I can imagine it's not so difficult. No offense Tedar obviously

John Tigges (00:53:40):
No offense whatsoever. It was because of his language. He promised her he would not use foul language. And if he did, he would give her money. Well, let's just say he lost that. Uh, and, and she made out like, like she robbed a bank. So

Peter O'Toole (00:53:55):
Did, did, was that for every word? Is that why she actually robbed a bank at the end? Yeah. She had that amount of money.

John Tigges (00:54:02):
She had that kind of amount of money, so yeah, it was, uh, and she couldn't have been happier. She love to this day. She loves him.

Peter O'Toole (00:54:10):
Ah, he's a great guy though.

John Tigges (00:54:13):
Oh,

Peter O'Toole (00:54:15):
What's your favorite film?

John Tigges (00:54:16):
Day off.

Peter O'Toole (00:54:22):
Ferries Bueller's day off utter. That's

John Tigges (00:54:26):
One, one of my favorite films. Um, along with every Halloween, I watched the Crow with Brandon Lee, the original. Okay.

Peter O'Toole (00:54:33):
So you got, so you got your Halloween movie. What's your Christmas movie, then?

John Tigges (00:54:39):
Christmas movie would be, uh, national lampoons. Christmas vacation.

Peter O'Toole (00:54:42):
Oh, cherry check. Yeah. We, we do watch that every year. Mean we've got people watch every year with the family, but yes, that is one of them,

John Tigges (00:54:51):
Of course, when, uh, so we go to my, uh, wife's cousin house and we tend to watch Die Hard as a Christmas favorite for some reason. I don't know how that became a Christmas movie. Come

Peter O'Toole (00:55:02):
On Christmas movie, isn't it?

John Tigges (00:55:04):
Yes, it is the, the, it may be the ultimate Christmas movie. Uh,

Peter O'Toole (00:55:10):
I it's, no, it's excellent. What, what is your pet hate? What do you really dislike people doing or things in life or something that's happening? What is your pet hate? Start with an, uh, cause you're just breaking up at the start of the sentences.

John Tigges (00:55:23):
Uh, yes. Uh, uh, um, my pet peeve, um, full sense of entitlement, I guess it's because I feel like I've worked hard to earn where I am. And so those that just show up and believe everything should be given to them really gets me going. Um, it's no, I mean, you may have certain opportunities, but then it's on you to make it happen. Yeah. That's, that's kind of a pet peeve. Um,

Peter O'Toole (00:55:58):
Yeah, that, that's a good one. Uh, I, I can see that if you support a football team that I support, I think a lot of our players seem to have that sense of entitlement. They are not delivering. Uh, what about, do you have any bad habits?

John Tigges (00:56:13):
Uh, bad habits is, um, I, I get that, that whole, uh, distraction thing very easily. I tend to be thinking of 10 to 15 things at all times. And this sometimes drives individuals that I need to get things accomplished crazy, cuz they're like, well, you just told me to do. Yeah, yeah. Forget that for the moment. and I just thought of this one and so it's yeah, that's a really bad habit. And then, um, my wife hates the fact that when I, I get nervous or a little bit upset, I chew the inside of my lip. And so she knows and says, you know, she'll look at me and be like, if you're angry, just say so I'm like, I'm fine. She's like, well then why are you chewing your lip? Uh, it's a tell, I guess it may not be a bad habit. It might just be, uh, if you play poker with me, look for the chewing and the lip.

Peter O'Toole (00:57:13):
I was just thinking of that when you've gambled on the wrong hand, just look for that. But now we know that you could be bluffing.

John Tigges (00:57:20):
Yes. There we go. Maybe I just put that out there.

Peter O'Toole (00:57:22):
That's not, that's not so good at that point. I, you, we are out time and I've got a few more questions. I really want to ask you first all, what do you do to chill out after, after a difficult day after, after off all of your, that you keep doing with your daughters or with work and losing your mentees as it were, what, what do you do to chill out at home?

John Tigges (00:57:45):
Uh, chill out for me is hitting the gym. I get to work off all the excess energy and stress. So my younger daughter, she has become a gym rat. So she, when even when I think I don't want to go, I go, um, so she ha does that. And then, uh, I play, I love to play basketball, been playing since, uh, I hadn't picked up a basketball until I was like 14 and it was just on a, someone needed a extra warm body. Um, and then I just kind of, cuz I was always a football player. I was playing soccer here in the states and this is why I still like it. And people think that's weird for an American. Um, I'm like, yeah, but I played the sport, but then I started playing basketball and it was a lot easier to find, you know, a couple of guys to play basketball than it was to find a few people to go and just to pick up soccer game. So,

Peter O'Toole (00:58:39):
Ah, soccer's still better. Sorry. That's a personal opinion. Uh, you, uh, do work now. We started to branch out that, you know, we've talked about teaching, mentoring, but you also thinking community wise networking and being responsible. So you now doing work with the ABRF. So do you wanna describe a, what the ABRF is? I, I, I know what we have the CTLs and actually I've been part of the international committee for ABRF as well, but you, you are more involved with that now. So go just in, just, just introduce ABRF and what you're doing at the moment.

John Tigges (00:59:14):
Okay. So the ABRF is the American Biomolecular Resource Federation, um, very fancy term, I guess, which is, um, a group that kind of, as you know, from your experiences, um, you know, cores in general, so or management and all the particular cores and I belong to the flow cytometry research group, um, of individuals, uh, that we put projects together. So you, you know, same thing as type writing grants, you write, you know, a grant to the society to see about, you know, what you'll actually fund with papers and minds manuscripts. So the thing I'm involved with them right now in is at a technology and technical pathway to a core facility. So working with local, uh, community colleges or, you know, universities to set up a program. So that individuals aren't just coming to a core saying, oh, well I did the biology requirements. It's going to a core having core skills. And that entire entails, which I worked, uh, with the, you know, the group to say, well, we need to add in there the management portion. So that's what we were saying. The individual who not only knows the technology, but knows the people skills knows the salesmanship knows how networking company relations can benefit, um, and understanding budgets. And you know, why can't I hire six people to work in the core? we'd be more efficient. Well, the budget says, no you will not. So, and also understanding how to work that budget to get what you actually need. So it it's a, it's a wonderful kind of thing that I'm, I'm happy to be, you know, involved in. Um, and also the group is working on how to get people to actually do titrations. And also what happens if you're sorting larger cells on your normal sorter. So, you know, people don't, they say there's, which I agree with a hundred percent people don't take in consideration that you're using a, a three micron bead to set up any kind of drop delay. But what happens when you start shoving things? The size of my head through a nozzle, even a, you know, a larger nozzle, how does that affect side streams and everything else? So, yeah, we use that analogy all the time, the size of my head and the brightness. I can describe nano cytometry by how the light hits off the top of my head.

Peter O'Toole (01:01:46):
So that's I, to you, you've got, uh, two laser lights going on as well. So you've got two, two,

John Tigges (01:01:52):
Uh, yes, we, we have two laser lights, so it it's good. I've got, I've got side scatter and backscatter going on. So

Peter O'Toole (01:02:01):
, I, I just going back to the finances side, uh, so many core stuff, don't like the finances ultimately. It's what empowers us. I, I actually think, uh, it it's it's yes, it's a pain. Yes. It it's difficult. Yes. It feels awkward. But I do think when finances are put properly into place, it can make a profound difference to how a facility can really operate. Cause you can justify, you know, users are justifying it. You don't have to have it cuz people say, oh yes, we want it. People are really putting their money. It's voting with their money. If they need that facility, they need that staff it's tremendously empowering. I think for the, a ARBF just like CTLs and actually even with the Royal microscopic Society, we even teach courses on facility management, uh, you know, for, for people wanting to go into this sorts of career path. Cause

John Tigges (01:02:57):
I saw that on LinkedIn for CTLs, that they have an actual, uh, couple of day intensive course on the, the business portions of a, of a core. Um, and I've always been interested in that. Like I said, when we started, I grew up in this or for better or for worse, I've learned, uh, business administration. I've learned, uh, legal, um, studies there because of contracts and you know, and just how things behind the scenes need to operate in order for a facility i.e. A small business to operate effectively. So, you know, I think that's, what's get lost sometimes in, in the research portion is that we are a small business. Um, you know, one of that mom and pop store in the mall, um, you know, you've got the big entities of, of the clinical and, and things that are bringing money to the hospital. And you've got your little portion of it as people are walking through and you're trying to get 'em to come into your little shop and stay relevant in, in, you know, financially sound. So it's an interesting, I always tell people that and they go, that's an interesting way to look at things. And I'm like, well, we are, if we don't have butts and seats, we're gonna fail. And so you need to figure out how can you build those relationships? And, and the other thing that has been relevant on the Purdue site even lately, and I've said it from beginning of time when I started this job, we are the bartenders of the science world. Um, I know things about people that maybe I should not know but here I am. Uh, and you know, I've been asked for advice on so many different topics, you know, that we could do an entire other session just on us going back and forth on what's the strangest thing you've ever been asked.

Peter O'Toole (01:04:49):
Yeah. The bartender, the agony aunt of, yeah, very much. So John, we are up to the hour. So I'm gonna say actually, I, I dunno you said the bartender, but I also say through listening to this and the diversity of your career, your job is I've also got the teacher, the businessman, the legal man, the accountants, the DJ / MC the family man, but maybe most importantly, the scientist, uh, that's there, John, it's been great catching up with you today. It really has

John Tigges (01:05:21):
Great. And I can't wait to actually be able to see you in person. Wow. Um, yes. So we can share, share some more stories in a pint together. Ah, um,

Peter O'Toole (01:05:31):
I know there's no point coming over on the RMS facilities meeting because you know it already, we need to think of another event we'll hook up soon. Uh, yes, but for those who are watching the Flow Stars do go back, watch the other ones in the other series that are out there as well. Some great speakers, including Alfonso, who you've heard, uh, a lot about today. Don't forget subs, forget to subscribe to the channels and John as always utterly. Brilliant. Thank you.

John Tigges (01:05:59):
Thank you too, Peter. It was an honor.