Aarati Asundi (00:12) Hi everyone, and welcome back to the Smart Tea Podcast, where we talk about the lives of scientists and innovators who shape the world. I'm Aarati. Jyoti Asundi (00:20) And I'm her mom, Jyoti. Aarati Asundi (00:22) And mom, I'm really curious if you've heard of the scientist we're gonna talk about today. His name is Roger Tsien. Jyoti Asundi (00:31) C-H-E-N? Aarati Asundi (00:32) T-S-I-E-N Jyoti Asundi (00:35) Hm. It rings a bell but I'm not able to place it. But I am pretty bad with names, you know that. Aarati Asundi (00:43) But you know what? This is exactly what this podcast is about because I know for a fact that you have worked his discovery or what he's known for literally every single day of your life. Jyoti Asundi (00:56) This is becoming more and more intriguing by the minute. Aarati Asundi (00:59) So Roger Tsien along with Osamu Shimomura and Martin Chalfie jointly all won the Nobel Prize for their work on fluorescent proteins. Jyoti Asundi (01:12) Oh! You are right. I do and I have extensively used fluorescent proteins. It is such a beautiful way to track cell-cell interactions, things happening within the cell. And you can tag proteins with different colors so that you can then have beautiful pictures of what various proteins are doing with respect to each other, where are they located with respect to each other, what happens when something is added, and so then they change and align differently, do things differently. Wow, I did not know that they were responsible for it. This is going to be so much fun. Aarati Asundi (01:58) And I also thought in order to add to the fun, we should play a game while we're going through this story. So every time I mention a Nobel Prize winner, we're gonna take a shot or a sip of water because this story is just littered with them. There are Prize laureates everywhere in this story. Jyoti Asundi (02:18) This is fantastic. So is work from a very high level. Aarati Asundi (02:23) Oh yes, almost immediately Roger is surrounded by amazing scientists and role models. and so he's basically set up for greatness from the beginning. We're gonna be well hydrated at the end of this story. Jyoti Asundi (02:38) That sounds good. I like- I like being hydrated. Okay. Aarati Asundi (02:42) So we're starting with his amazing father, his father, Hsue Chu Tsien, was an aeronautical engineer, and he was serving in the Chinese Air Force when he met Roger's mother, Yi Ying Li, who was a nurse. They started a family there and they had their first son named Yongyou. Then during World War II, Hsue Chu was ordered to go to the US as a liaison to try and get more aid for the Chinese military. But because there's a war going on, he had to zigzag all over the world in order to avoid enemy forces. And so by the time he landed in the US, the war was over. Japan had surrendered, and now Hsue Chu is in the US. And he realized that since China was on the losing side of the war, if he went back there, it would be tough to build a life because China would be trying to rebuild from a war that they had lost. Jyoti Asundi (03:44) Yes, that's right. Aarati Asundi (03:47) And so he instead made arrangements for his wife and his young son, Yongyou, to come and join him in the US. Jyoti Asundi (03:55) Oh! I'm that it happened it seems to have happened fairly easily. Because at this point would have thought that the US would think of China as the losing side and therefore the other, the other basically. Aarati Asundi (04:09) The enemy. Yes Jyoti Asundi (04:10) The enemy exactly. And yet they allowed a Chinese family to be reunited. Aarati Asundi (04:17) Yes, so they were able to immigrate, but to your point, life was still a struggle for them exactly because of that. Hsue Chu had a really hard time finding a job as an aeronautical engineer because he couldn't get the security clearances that he needed to work in that field. Jyoti Asundi (04:34) Yeah, that actually makes a lot of sense. Wars leave behind a lot of mistrust especially if they will be looking different, they seem to be different. I can see the human prejudice coming into play here. Aarati Asundi (04:47) Yes, very strongly. And so instead, in order to get by, he started a small import export business and then later started a engineering consulting company which, you know, allowed them to kind of get by. They weren't super wealthy or anything, but they were managing. The couple had two more children, son who they named Yonglo and then their third son, Yonchien, was born on February 1st 1952, in New York City. And he's the star of our show today. Jyoti Asundi (05:12) Ok. So Roger Tsien was actually born Yonchien? Aarati Asundi (05:27) Yes. So around this time, their first child, Yongyou, was just starting school. And the school officials told him he needed to pick an American name. Jyoti Asundi (05:41) Okay. Oh this would not fly today. Aarati Asundi (05:45) Yeah, yeah. Jyoti Asundi (05:46) Okay. All right. But a small point here one of the things I have as I have navigated my way in America for so many years is that a lot of Indians maintain their name. If they change it at all, it is extremely small change. So for example, you know, somebody be like Raj Kishore or something and they'll just shorten it to Raj. Or something like that, you know. So something... Aarati Asundi (06:12) Yeah. Which is what you do in America also anyway, you know? Jyoti Asundi (06:16) Anyway, correct. Aarati Asundi (06:16) Like, yeah, like Jennifer becomes Jen. Jyoti Asundi (06:20) Exactly. However in contrast I always noticed that people who came in from China would change their name drastically. Exactly like you say they would say Yongchien or something like But call me Roger. And I would just think to myself, but where is the connection? Like that doesn't make any sense to me. So maybe this is it. Since the Chinese immigrants came in much earlier, they had been conditioned to do this. Like you come into America and you change your name because your name is not pronounceable to the rest of the Western world. So you do it like this. But Indians came a bit later where there was more openness already building in America. People were like, hey, I appreciate your culture. I embrace it. Tell me your name. I'm going to work hard my hardest to pronounce it correctly. Just tell me how to do it. And that's what I faced always. I would I would tell my name and then people would say, me again how to say it and then I would say it and then we would work it out. So this and I always wondered why, why, why is this happening? This gives me a big clue here. Aarati Asundi (07:33) Yeah. So in this case, it was kind of random how they came up with their names. So the oldest child Yongyou was just told he needed to pick an American name, and he said he wanted to be called Dick. And the school explained to him that that was a nickname for Richard, so he became Richard. Their middle son became Louis, and then when asked what Yongchien's American name should be, Dick said it should be Roger because he liked the actor Roy Rogers, who was a cowboy film star. Jyoti Asundi (08:09) Okay, yeah Roy Rogers is a hero. I want my little brother to be a hero. Okay, yes. Aarati Asundi (08:12) Yeah. Yes. And so then their Chinese names became their middle names or some translation of their Chinese name. Jyoti Asundi (08:19) Okay. Some some variation. Okay. Aarati Asundi (08:22) Yeah. So Roger's name now is Roger Yonchien Tsien. Very early on, Roger got interested in chemistry, particularly in experiments that were colorful. He shared two examples in his autobiography. The first is what he called silica gardens, which is where you drop a colorful metal salt into sodium silicate or liquid glass, and the salt forms this kind of jelly-like coating, which would then grow long tendrils growing upwards. And so over time you get kind of this weird colorful underwater garden looking thing. Jyoti Asundi (09:06) Oh interesting! Aarati Asundi (09:07) Yeah. I'll post a video on the website so that people can go and see it. It's really cool looking. Jyoti Asundi (09:14) Yeah! This this sounds like a fun little experiment for any middle schooler or even elementary schooler to do. Aarati Asundi (09:20) Yeah, so he really liked it. And depending on what color salt you use, you get different colored, seaweeds or tendrils growing upwards. Jyoti Asundi (09:27) Yes. Aarati Asundi (09:29) The second experiment he enjoyed a lot was creating a strongly basic solution of potassium permanganate, which has a very strong purple color. And when you pass this solution through a paper filter, the potassium permanganate steals electrons from the cellulose in the paper. And this changes the way that the permanganate absorbs light. So once it gains an electron, potassium permanganate turns green So you're pouring in this purple liquid and it comes out green on the other side. Very cool. Jyoti Asundi (10:04) Oh my goodness! Oh I wish I knew these experiments when I was in middle school it would have been fun to do this. Aarati Asundi (10:12) Yeah. And actually Roger, like later after he won the Nobel Prize and he was talking about these experiments that are what piqued his interest in chemistry, people are like, we've never heard of that and he's like, "What? You've never heard- you've never done this before? Let me show you." And he like demonstrated it. So he was pretty surprised that people hadn't heard of these experiments also. They're really cool. Jyoti Asundi (10:32) Yes, yes. Aarati Asundi (10:34) And actually potassium permanganate is really cool. I was looking into it and it's called the chemical chameleon because if you keep on giving it electrons and keep on doing these redox reactions, the potassium permanganate will turn into manganese dioxide, which has a brownish yellow color. And then eventually if you keep going, keep on doing a redox reaction, keep on giving it electrons, you'll end up with just the manganese ion, which has a very pale pink or clear color. Jyoti Asundi (11:04) Oh wow. Aarati Asundi (11:04) So you can transition through this whole- from purple to green to yellow to pinkish. Very cool. Jyoti Asundi (11:12) This is fabulous. Chemical chameleon. Aarati Asundi (11:14) Yes. Very cool. Jyoti Asundi (11:17) Yes. Aarati Asundi (11:18) In 1950, when Roger is seven, his father started working for the Radio Corporation of America, or RCA. And so the family moved to New Jersey. His parents found a new housing development that was being built in Livingston, which was perfect in terms of his dad's commute to work, and it was in a good school district for the three boys. But the developer refused to sell them a home, saying "They could not permit Livingston to become a Chinatown, nor could they afford the likelihood that other customers would refuse to buy houses next to a Chinese family." Jyoti Asundi (11:56) Oh! Discrimination raises its ugly head. Okay. Aarati Asundi (11:59) Yes. So the Tsiens eventually had to appeal to the governor of New Jersey, Robert Meyner And he wrote a letter to the developer saying that what they were doing was illegal racial discrimination. They couldn't refuse. Jyoti Asundi (12:13) Yes, yes. Yeah, I thought there are laws against it. Yeah. Aarati Asundi (12:18) Yes. And so finally the developers compromised by selling them a house that was completely surrounded by other houses that were already sold. Jyoti Asundi (12:28) Yes, because then nobody else can refuse to buy a house next to them. Aarati Asundi (12:33) Yes. Roger was a very scrawny, non-athletic kid who suffered from asthma, so he spent a lot of his time indoors. But he really tried to integrate himself into American culture. He didn't take any interest in learning Chinese traditions or the language or eating their authentic foods. So I found that interesting. Jyoti Asundi (12:55) Okay. But I can- I can imagine that with the kind of discrimination they're facing, it's not making you unique and memorable, it's making you a target. And I can see how as a young child you don't want to waste your energies on that. You just want to kind of go with the flow. Aarati Asundi (13:15) Yeah, and like you were saying, like I'm sure he already stuck out because he obviously looked Chinese and so, you know. Jyoti Asundi (13:21) Yes, yes. Aarati Asundi (13:22) But he did keep up his intense interest in chemistry, turning his basement into a lab. His dad eventually moved jobs again and started working for Exxon Research and he would secretly bring him home chemicals and glassware or help him special order equipment that he needed for doing his experiments. Jyoti Asundi (13:43) This is how he had access because that was a kind of question bugging me at the back of the head. That solves one mystery for me. Okay. Aarati Asundi (13:53) In high school, he entered a summer program sponsored by the National Science Foundation, where he was assigned to work in a chemistry lab at Ohio University under Professor Robert Kline. He worked on a project involving thiocyanate, which is a compound that's used to neutralize cyanide. So Jyoti Asundi (14:12) Okay. Aarati Asundi (14:12) So like if we humans ingest small amounts of cyanide from certain seeds or environmental toxins, our body converts it to thiocyanate, Jyoti Asundi (14:23) Ah I see! Aarati Asundi (14:23) ...which is not toxic and... Jyoti Asundi (14:24) Not as toxic. Okay. Aarati Asundi (14:26) Yeah. So Professor Kline was interested in whether thiocyanate could bond to different types of metals at the same time. And so Roger's summer project was a lot of making different compounds out of thiocyanate and taking measurements. The compounds he made were not very well defined and his results were inconclusive. But the following year, he entered a nationwide competition called the Westinghouse Science Talent Search. And as Roger says, "For lack of any alternatives, I wrote up my Ohio University project, trying my best to draw some conclusions from a mess of dubious data." Jyoti Asundi (15:10) Been there, done that. This is what research is all about. Even now it's not necessarily as a high schooler, even after years and years you conduct experiments and.. Aarati Asundi (15:24) It's just yeah, so confusing. Jyoti Asundi (15:24) ...you have to just look at the mess of the data and say, " Oh there is so much noise. How do I draw a clear conclusion out of this?" Aarati Asundi (15:33) Yeah. And so he tried drawing some sort of conclusions in order to, you know, because he's entering a science competition, he wants a nice story. But even Professor Kline was not really willing to back up any of the conclusions Roger drew. And Roger was like, "Yeah, fair enough. Like, this is not great." Jyoti Asundi (15:50) Yes. I understand, yes. Aarati Asundi (15:53) Yeah. But he entered this science competition and you know, he was like not having very high hopes because he was like, you know, my god, everyone else's project is so much better and cleaner than mine and so nice. And he was also very intimidated by the fact that the competition was judged by a Nobel Prize laureate, the first one in our story, Glenn Seaborg, who won the prize for discovering 10 elements on the periodic table, numbers 93 to 102, all of which are radioactively unstable. Jyoti Asundi (16:30) Glenn Seaborg, the one who found ten unstable radioactive elements. And I'll go for my water too. Hang on. Aarati Asundi (16:37) Yes, take a shot. ⁓ Take a sip of water. Jyoti Asundi (16:38) There it is. To send to Glenn Seaborg. Aarati Asundi (16:40) Yes. Nevertheless, Roger went on to win the entire competition, to his intense surprise, taking home a $10,000 scholarship prize. Jyoti Asundi (16:54) That is big money in those times. It probably funded his entire college education in those days. Aarati Asundi (16:59) It might have, or a big chunk of it. Jyoti Asundi (17:02) Good for him. Wow. Aarati Asundi (17:02) Yeah. But the thing that gave him the most satisfaction was that when he brought home the prize, the housing developers that hadn't wanted to sell their family that house started using Roger's award picture in their advertisements to show what a good school system they were part of. Jyoti Asundi (17:22) Are you serious? Are you serious? Aarati Asundi (17:24) Isn't that great? Jyoti Asundi (17:26) Hypocrisy at its best. This is really hilarious. They not only had to eat their own words, they had to advertise it to the to the entire world to let people know "Okay forget that. We never said 'no' to this wonderful, wonderful family who wanted to join our school system, who is so amazing." Aarati Asundi (17:50) Yes. Jyoti Asundi (17:52) This is karma at its best. This is what karma is about. Aarati Asundi (17:54) Right? This is this is hundred percent I was like, that's karma. Yes. Jyoti Asundi (17:58) Yes, this is karma. Yes, good, very good. Aarati Asundi (18:02) Roger now graduated high school and he is facing a choice of four colleges. His choices are Columbia, MIT, Caltech, or Harvard. What to do? What to do? Yeah. Jyoti Asundi (18:15) Whoa, can't go wrong. Well, the only thing I can say is with those kind of choices, it doesn't matter. You can close your eyes and toss a coin or spin a wheel or whatever you want and you can't go wrong. Aarati Asundi (18:30) Yep. So here's how he made his decision. His dad told him not to go to Colombia because recently there had been a lot of student protests there against the Vietnam War, and so all the unrest made him uneasy. And Roger also was like,"Eh, Colombia's too close to home anyway." So that's out. Both of his older brothers had gone to MIT, and so Roger's like, "Well, I wanna be different. I don't wanna do that." Jyoti Asundi (18:58) Yes, because everybody will remember, "Oh you're Richard's younger brother!" you don't wanna be in that situation, always being compared. Aarati Asundi (19:04) Yeah, he didn't want to be compared. Jyoti Asundi (19:06) No, you're always- you're setting yourself up to compared. It's not fun. Aarati Asundi (19:11) Yeah, so MIT was out. He was very drawn to Caltech, but ultimately decided against it because Nobel Prize winner Richard Feynman was no longer teaching intro to physics there. Jyoti Asundi (19:24) Whoa! And that brought Caltech down in his eyes. ⁓ wow, wow. Aarati Asundi (19:28) Yes. Yeah. And so he's left with Harvard. And overall he had a great time at Harvard. The culture was great. There was a lot of diversity in both students and classes. Ironically though, he hated his chemistry classes. Jyoti Asundi (19:47) Oh no. Aarati Asundi (19:48) And I think specifically it was organic chemistry, which was, you know, the thing that he really disliked. And I can understand that. Like everyone in my major and when I was doing biochemistry as my major, everyone was like, "Oh no, the dreaded O-Chem. We all hate O-Chem." Jyoti Asundi (20:05) You know, O-Chem I think gets a bad rap because people who teach it do not know how to teach it correctly. I think you faced that as well when you were learning O-Chem. Aarati Asundi (20:17) I had a couple of good teachers, but I had a couple of bad ones too. Jyoti Asundi (20:21) However, I think you did best once you were able to get that kinetic model with which you were able to work. Aarati Asundi (20:29) Oh yeah, a very hands on learner, yeah. Jyoti Asundi (20:31) And I think O-Chem is one of those need to have multiple media integrated into teaching. Aarati Asundi (20:39) Yeah, and I think a lot of the professors immediately jumped to teaching us mnemonic devices to remember things rather than like actually how nature works and I'm terrible with that kind of thing. Jyoti Asundi (20:49) Yeah, no, that's, that's just memorization without understanding at all, and that doesn't work either. Aarati Asundi (20:54) Yeah, so he hated his chemistry classes and he started exploring other courses in molecular biology, quantum mechanics, astrophysics, but eventually he landed on neurobiology... Jyoti Asundi (21:09) Okay. Aarati Asundi (21:09) ...in part because of two professors, Professor David Hubel and Torsten Wiesel, who- Both of them would go on to share the Nobel Prize in nineteen eighty one, along with Dr. Roger Sperry, for understanding how light signals from our eyes are processed into visual information in the brain. Jyoti Asundi (21:31) Wow. Is it enough to just take a sip of water? Do we have to geneflect also? Aarati Asundi (21:32) Yes. Yes three small sips maybe. Yeah. Jyoti Asundi (21:38) Should we should we also do some geneflecting here? I don't know. Aarati Asundi (21:41) Yes. So these two Nobel Prize laureates who would eventually be... like at the time that they were lecturing and teaching Roger, they hadn't won the Nobel Prize yet, but they were just so enamored by neuroscience and the way that they lectured to their students, saying like neuroscience is the next frontier and we need to understand how the brain works. And Roger was just completely taken in by that and he was like, Yes, that sounds fascinating. Jyoti Asundi (22:09) Yeah. Makes sense. If they went on to win the Nobel Prize, obviously they were highly passionate about it and they were able to communicate their passion for their subject to their and basically ignite the same passion in their students as well. Aarati Asundi (22:26) Yes. So Roger decided I'm gonna go into neuroscience. So he asked Professor Hubel if he could join his lab, but he wasn't taking on any undergraduate students. And so he suggested that Roger apply to the lab of Dr. Nelson Kiang instead, who worked on auditory neurophysiology or how sound signals from the ear go to the brain and are processed. So Roger did that, and then for graduate school he wanted to continue working in neuroscience. So he asked Professor Hubel and Professor Kiang which were the best schools for neuroscience. And they told him, Cambridge or Cambridge. So either stay at Harvard in Cambridge, Massachusetts, or go to Cambridge in the UK. Jyoti Asundi (23:12) Cool, what a nice way to put it. Aarati Asundi (23:14) And so Roger was like, "Oh well, I'm up for a new adventure. Let's go to Cambridge in the UK because I've done Harvard, let's go see something new." Jyoti Asundi (23:24) See what something else has to offer, yeah. Aarati Asundi (23:26) So he applied for the graduate program there and he got in and he was told that he would be working under Dr. Richard Adrian, whom he had never heard of. So he called his older brother Dick, was now a professor of cardiac electrophysiology at Yale. Jyoti Asundi (23:45) Wow. Aarati Asundi (23:45) Yeah. And Dick said, "Oh yeah, I know Professor Adrian. He is the son of Dr. Edgar Adrian, who is a Nobel Prize laureate. Jyoti Asundi (23:58) Wow, he is in such a rarefied atmosphere. Aarati Asundi (24:03) Yes. Jyoti Asundi (24:03) This- how often does this happen to anybody in the world? Aarati Asundi (24:06) I know. I was not kidding about this. Jyoti Asundi (24:08) Just wherever he goes... yeah, wherever he goes, he's surrounded by these extremely knowledgeable, passionate scientists. Amazing. We forgot our sip. Aarati Asundi (24:18) Yes. So yeah, take a sip. So Dr. Edgar Adrian won the Nobel Prize for understanding the electrical activity in the brain. And so Dick told Roger that Richard Adrian, the son, was a skeletal muscle electrophysiologist. And Roger was like, "Oh no, I don't want to study muscles. Muscles are backwater research. I wanted to work on the brain." Jyoti Asundi (24:46) Okay. Aarati Asundi (24:47) But Dick told him he would be okay. Professor Adrian would probably let Roger choose his own thesis topic, so he could probably work on the brain if he wanted to. So Roger decided to take a chance and he went to Cambridge. And when he first met Professor Adrian, Professor Adrian asked Roger, are you Roger Tsien? And Roger was surprised that Adrian had pronounced his last name very correctly. Jyoti Asundi (25:14) Yeah. Aarati Asundi (25:15) Professor Adrian then asked if it was true that Roger thought that muscle research was a backwater field of research. Jyoti Asundi (25:21) No, wait, wait, how did he find out? How did he find out? Aarati Asundi (25:25) And Roger realized that his brother Dick had met with Professor Adrian over the summer and spilled the beans. Jyoti Asundi (25:32) His brother sold him out? Aarati Asundi (25:34) Yes. Jyoti Asundi (25:35) That is so mean. Aarati Asundi (25:37) That's such an older brother thing to do, I feel like. Jyoti Asundi (25:40) That is so terrible to set him up in his brand new place where he's going. He doesn't know anybody. He's leaving one country to go to another and his brother sets him up with his new professor. Aarati Asundi (25:53) Yes. Jyoti Asundi (25:54) I would want to- I would want to pound him into the sand when I came back home for Christmas. Aarati Asundi (25:58) Yeah. I would want to kill him. Yeah. Jyoti Asundi (26:00) I absolutely would. Aarati Asundi (26:02) Right? But Professor Adrian was very gracious about it. Jyoti Asundi (26:07) Of course. That's I mean, I'm sure that's why the brother able to open up as well. I- I get that. I totally get it. I'm sure the his... Aarati Asundi (26:15) Yeah. I think the older brother was just kind of being mischievous and was teasing Professor Adrian like, "Hey" Jyoti Asundi (26:18) Yeah, they're probably the two older guys... Aarati Asundi (26:22) They're probably good friends. Jyoti Asundi (26:24) You know, are probably rolling their eyes at young wannabe scientist who is like, "Ew, muscle research is so ew. I wanna be in the brain." And they're probably rolling their eyes and making fun of him typical older brothers and friends do. But not fair! Not fair! Aarati Asundi (26:41) Yes. Not nice. Yes. But Professor Adrian was gracious about it, and he confirmed that he would even let Roger transfer to a neurophysiology advisor for his thesis if he wanted to. But Roger quickly realized that actually neuroscience research meant doing a lot of electrophysiology work, which he did not enjoy. So basically, when a neuron gets excited enough, sodium channels on the membrane of the neuron open up, allowing a huge influx of electrically charged sodium ions into the neuron, which creates an electric pulse called an action potential. The action potential starts in the main cell body of the neuron and then travels down the axon and ends up at the axon terminal where the neuron forms synapses with other neurons. Jyoti Asundi (27:33) And thus the signal gets transmitted. Aarati Asundi (27:35) Yes. Here in the axon terminal, the action potential causes calcium channels to open And calcium ions enter the neuron. And this signals a release of neurotransmitters that go and signal other neurons or other parts of the body. Jyoti Asundi (27:51) Okay. Aarati Asundi (27:52) And so the way that Hubel and Kiang at Harvard were doing neuroscience was by putting a micro electrode into the brain of the animal, and then they would record the activity of an individual neuron. So they would give the animal some sensory stimulus and then see if that neuron, that individual neuron that they were measuring, lit up in response to the sensory stimulus. Jyoti Asundi (28:15) Got it. Yeah. So you hone in on your most likely neuron up in the brain and give a signal let us say on the foot or the finger or something like that or create a create some sort of a... Aarati Asundi (28:27) Well, they were doing visual things, so they would probably give the animals something to look at, yeah. Jyoti Asundi (28:32) Yes, give something to look at and see if the neuron lights up, but it could potentially be the wrong neuron that you're chasing, and so you have to go and do the same thing again with your next neuron now. And so on and so forth. Yeah. That okay, that makes sense. Aarati Asundi (28:42) Mm-hmm. Yeah, and you had to do that like a bunch of times until you came up with a pattern or some network that made sense. Jyoti Asundi (28:50) That's right. Okay. Aarati Asundi (28:52) And Roger was like, that is so tedious. Jyoti Asundi (28:55) Absolutely! Aarati Asundi (28:55) That's so slow. Yeah. we know that the brain is super complex, trillions of neurons all firing and signaling each other. And so he thought, what's a better way to do this? And he said, "Ideally, one would stain the neuron with a dye that would visibly light up or change color whenever and wherever a neuron fired an action potential." Jyoti Asundi (29:20) Ah! There! That's the idea coming through. Okay. Yes, yes. Aarati Asundi (29:24) Yeah. So in 1972, in graduate school, he decided to try and design a dye specifically to see neuronal activity. He thought the best way to do would be to either create a dye that would become visible when the sodium channels opened up when the neuron initially got excited, or maybe a dye that responded to the electrical changes that happened across the neuron's membrane. Jyoti Asundi (29:51) Got it. So the dye is inert, but then it lights up because of its interaction with the sodium channel or because now it is being stimulated by the electric impulse. Aarati Asundi (30:02) Exactly. Something like that is what he's thinking. But either way, in order to do this, that meant he had to study organic chemistry, which were the classes that... Jyoti Asundi (30:11) Back to organic chemistry, which he hated it for probably good reason, okay. Aarati Asundi (30:16) Yes. But he was very determined and so he found a mentor, Dr. Ian Baxter, in the chemistry department to help him. And he realized that actually if he was studying organic chemistry for his own purposes, he enjoyed it a lot. So kind of like back to what we were saying, you know, like if you teach it the right way, you give some a student the proper motivation, they will learn. Jyoti Asundi (30:38) I think that's really what's missing sometimes with these complex teachings. You can't just go on droning about something, draw some structures on a blackboard and expect everybody to pick it up. Aarati Asundi (30:49) Yeah. So although he was able to create molecules that did bind to sodium channels, he wasn't having much success with them acting as an indicator of whether the neuron was getting excited or not. But then he read about a new dye, arsenazo III, that was able to bind to heavy metals and calcium. And he was like, hey, that could work too, because once the action potential reaches the terminal, calcium ions come in and trigger the release of the neurotransmitters. Jyoti Asundi (31:18) That's right. Yes. Aarati Asundi (31:22) So maybe targeting calcium is the better way to go. Jyoti Asundi (31:05) That makes sense because the interaction with sodium is not creating that different signal that you're looking for. Aarati Asundi (31:31) Mm-hmm. Yeah, and so maybe time to switch targets. And actually there are plenty of dyes, hundreds of dyes that could bind to calcium. The problem was, though, that inside the cells, these dyes would also bind to magnesium ions, which were about four times higher in concentration than calcium ions. Jyoti Asundi (31:51) Yes okay. Aarati Asundi (31:53) And so what he did was very clever. He found another molecule called EGTA, which was a colorless buffer, but it had selectivity for calcium, meaning that it strongly preferred to bind to calcium rather than magnesium. And then with a bit of playing around, he found a way to kind of combine EGTA's selectivity with a dye to make a new molecule called BAPTA. Jyoti Asundi (32:23) Ah! BAPTA is well used today. Aarati Asundi (32:26) Yes. Jyoti Asundi (32:26) Wow. I didn't realize that's how it came about. Aarati Asundi (32:30) So his version of it was a bit rudimentary and later other scientists took that concept and refined it to make other versions and that's probably actually what we use today. But that initial concept, it was the first dye that could selectively bind to calcium inside a cell and therefore tell you whether a neuron was excited or not. So it was huge. Jyoti Asundi (32:52) That's right. Yes, yes, that I can see this. Wow, exciting. _______________________________________________________________________________________ Grow Everything Podcast Advertisement Aarati Asundi (33:34) Hi everyone, Aarati here. I hope you're enjoying the podcast. If so, and you wish someone would tell your science story, I founded a science communications company called Sykom, that's S-Y-K-O-M, that can help. Sykom blends creativity with scientific accuracy to create all types of science, communications, content, including explainer videos, slide presentations, science, writing, and more. We work with academic researchers, tech companies, nonprofits, or really any scientists. To help simplify your science, check us out at sykommer.com. That's S-Y-K-O-M-M-E-R.com. Back to the story. _______________________________________________________________________________________ Aarati Asundi (34:22) So that was basically his PhD project. You know, he did that for his PhD then he stayed at Cambridge for his postdoc. And during this time he started collaborating with another new faculty member in the physiology department, Dr. Timothy Rink, who was also interested in making calcium selective electrodes. Jyoti Asundi (34:43) Okay. Aarati Asundi (34:43) They became really good friends, and Tim would invite Roger over to his house and you know, invite him to parties where his wife was there. And in 1973, at a Christmas party, Roger met Tim's sister-in-law, Wendy Globe, and they hit it off immediately. They started dating kind of secretly at first, and then several months later, when Tim and his wife found out, they were shocked at how well they had managed to set up these two without even meaning to. Jyoti Asundi (35:15) Meaning to. Inadvertent matchmakers. Aarati Asundi (35:20) Yes. Yeah. So after his postdoc fellowship was up, he tried his best to get a job in Britain because he wanted to stay close to Wendy. But finding a research position at the time was very difficult. So he turned back to the US. But there also he had a hard time. And the reason was kind of interesting. He was an interdisciplinary scientist with expertise in both biology and chemistry, which was weird for the time. Jyoti Asundi (35:52) They are unicorns today who are highly prized. People recognize interdisciplinary is the way to go and- and break down those boundaries of science. Aarati Asundi (36:05) Yes. But back then biology departments were rejecting him because he was too much of a chemist, and chemistry departments were rejecting him for being too much of a biologist. Jyoti Asundi (36:15) They wanted a purist. Aarati Asundi (36:16) Yeah. Jyoti Asundi (36:17) That's not how science works anymore. Aarati Asundi (36:19) He was way ahead of his time. Finally, though, through some network connections, he was able to land a professorship at Berkeley, but his startup funding package was extremely low, just a few thousand dollars. So to set up his lab he had to rely on old equipment or create some very strong justification for getting new things. Another professor who worked in a lab next door donated an old fume hood to him, and so for the next seven years all the chemical reactions that were done in the lab had to be done in that single donated fume hood. But still, he was able to make improvements on his calcium ion indicators and synthesized a sodium ion indicator, so back to the sodium channels. Jyoti Asundi (37:07) Oh back to sodium, okay. Aarati Asundi (37:09) Yep, and take images of both of these indicators inside individual living cells. And in 1982 he married Wendy, and so she came to the US to live with him. Jyoti Asundi (37:21) Okay. Aarati Asundi (37:22) But after several years he outgrew his lab at Berkeley. Not only was he constantly struggling with limited and outdated equipment, but he also felt like his research was being boxed into just imaging ions, like just looking at calcium ions or sodium ions, and he wanted to look at bigger biochemical molecules and their interactions. Jyoti Asundi (37:45) Yes. Aarati Asundi (37:46) He was particularly interested in cyclic AMP, which acts as a messenger inside cells. Jyoti Asundi (37:54) Yes. Aarati Asundi (37:55) So, once a signal comes from outside of the cell, and that signal could be either a neurotransmitter or a hormone from some other stimuli, cyclic AMP runs around inside the cell, activating pathways that regulate the cell's response. And these could be pathways that are metabolic, gene expression pathways, enzymatic pathways, all sorts of things. Jyoti Asundi (38:18) Okay. Aarati Asundi (38:19) And actually in 1971, Dr. Earl Sutherland had won the Nobel Prize discovering that cyclic AMP worked as a messenger inside the cell as a response to epinephrine. So it's a hugely important molecule. Jyoti Asundi (38:33) We have to take a sip now. Aarati Asundi (38:35) Yes, because I had to mention. Jyoti Asundi (38:37) Sutherland. Aarati Asundi (38:38) Mm-hmm. And so in 1989, Roger and Wendy moved to UC San Diego, where he was able to get a much bigger lab and he was able to, you know, pursue this line of inquiry. At UC San Diego, there also happened to be another scientist named Susan Taylor, and she was an expert on cyclic AMP and its role in activating protein kinases. Jyoti Asundi (39:08) Okay. Aarati Asundi (39:09) So really how cyclic AMP works is once a signal is detected from outside the cell, cyclic AMP goes and binds to a protein kinase, usually protein kinase A. Jyoti Asundi (39:23) Okay. And protein kinases are the ones who actually move the phosphate around on the various molecules, to either activate or inactivate them. Aarati Asundi (39:33) Yes. So protein kinase actually is made up of two smaller subunits. So once cyclic AMP comes, it binds to one of the subunits of protein kinase, but the other subunit goes and starts, you know, phosphorylating things and dephosphorolating things and turning on and off enzymes, altering gene expression, metabolism, all of that. So cyclic AMP is just acting as the secondary messenger that once the cell senses something, cyclic AMP goes and says to protein kinase, hey, the cell has sensed something. Jyoti Asundi (40:10) It's really like a relay race. Like out there, all the way out there came a signal, a hormone or something. And somebody on the cell surface got tickled. And tickle then goes in and is transmitted over to the cyclic AMP, who is very agile and he goes running around, bounding away goes to the protein kinase, which is more internal to the cell. And now it's like the protein kinases now "oh I need to activate X pathway or Y pathway. And the way the pathways get activated or deactivated is through the phosphorus that is attached or removed." So that's how yeah, that's how it's working. Aarati Asundi (40:49) Yes. And so Roger comes up with a really cool now knowing this about how protein kinase and cyclic AMP interact. He thought instead of trying to add a dye or indicator to cyclic AMP, what if we add an indicator to each subunit of the protein kinase? So he added a green indicator called fluorescein to one subunit and a reddish dye called rhodamine to the other. Jyoti Asundi (41:20) Yes. Aarati Asundi (41:21) And so now when you look at the protein kinase, if it's intact, if both subunits are together, because the green fluorescein and the red rhodamine are so close to each other in space, you get a phenomenon called fluorescence resonance energy transfer or FRET. So any light that's coming from the green fluorescein gets absorbed by the rhodamine and is emitted as red light. So when the two pieces are together, you won't see any green light because it's all being emitted as red light from the rhodamine. Jyoti Asundi (41:56) Okay, due to the fret phenomenon, yes. Mm-hmm. Aarati Asundi (41:58) Yes. But when cyclic AMP comes along and breaks apart the protein kinase, then the two dye indicators will separate from each other because the subunits have separated. And so now you will see both red and green light in different places in the cell. Jyoti Asundi (42:14) Yeah got it, got it. Aarati Asundi (42:17) And so this not only tells you whether the protein kinase has been activated by cyclic AMP or not, but by measuring the ratio of red to green, you can also tell how much of the protein kinase has been activated. Jyoti Asundi (42:29) It is quantifiable. Okay. Yes, yes. Aarati Asundi (42:32) Yes. And so although this was great, they got some good papers out of this, they were able to publish, this process still had its drawbacks. It was very labor intensive. You had to purify the two subunits of protein kinase so that you could attach the dyes in a way that did not destroy the function of the subunits and then inject those two subunits back into living cells. And so Roger was like, we need to get away from using dyes if possible. They're such a pain. Instead, what if we could fuse the proteins we're interested in with a naturally occurring fluorescent protein? That would be ideal. Jyoti Asundi (43:14) Yes, we are inching closer to where we are today. I can see it. Okay. Aarati Asundi (43:19) Yes. And so he started turning to examples of naturally occurring fluorescent proteins in nature and mostly he was looking at bioluminescent algae, jellyfish, fireflies, fungi. Roger's like looking at all of these and studying how their bioluminescence came about, what proteins were causing their bioluminescence and how could we co-opt these for our own research purposes? Jyoti Asundi (43:49) Absolutely. Mm-hmm. Aarati Asundi (43:51) Then in 1992 he came across a paper from Dr. Douglas Prasher at the Woods Hole Oceanographic Institution in Massachusetts. Dr. Prasher had successfully been able to clone a bioluminescent protein from the Aequorea Victoria jellyfish called green fluorescent protein, or GFP. Yes. Jyoti Asundi (44:13) GFP. I was I was biting my tongue to not say GFP because I knew I would preempt you. So I did not want to say it. Don't say GFP. Aarati Asundi (44:24) Yes! Here it is. Jyoti Asundi (44:25) Green fluorescence protein. How many beautiful images they have created with that GFP. Aarati Asundi (44:29) Beautiful. Yes. So GFP had first been isolated and studied back in the 1960s by Dr. Shimomura. But no one had looked at whether it could be used in other animals or cells. So then in the late 80s, Dr. Prasher wanted to take up the study of GFP, but the NIH and NSF both declined giving him grant funding. Isn't that crazy? Jyoti Asundi (44:59) Sounds crazy today. Aarati Asundi (45:02) Yes. Jyoti Asundi (45:02) It's like one of the fundamental backbones of research today. It's so cost effective for ongoing research to use GFP. You have the sequence in every lab, every cloning lab has the GFP sequence ready to go to tag on to anything that people wanted to tag on to. And the highest institutions of science in America are like, "Nah, this is not interesting I don't want to fund it." It reminds me that lady from Alaska, Sarah Palin, was scoffing at research on fruit flies she was scoffing and making fun of it, "Who wants to study fruit flies?" Without the least it was it was obvious she did not understand that it was the model organism on which we could base a lot human complexity could be decoded using this... Aarati Asundi (45:52) And that's bad enough. I mean she's a politician, which doesn't give her a pass at all, but at least she was a politician. This is like the NIH and the NSF saying this GFP is not worth researching and it sounds... Jyoti Asundi (46:05) Incredible, incredible. Aarati Asundi (46:07) ...just crazy to hear that today. Jyoti Asundi (46:10) Yes, yes. I'm glad it was documented somewhere that they rejected these grants. From the way you have set up the story, it is pretty obvious that this is going towards a Nobel Prize. And so I'm glad that the naysayers were documented as being naysayers. Aarati Asundi (46:21) Yes. I think it's important to point it out. Jyoti Asundi (46:29) Yes, it's like you made a foolish decision before. I cannot trust you anymore. Aarati Asundi (46:33) So the people who are the heroes of the story who did give him funding are, surprisingly, the American Cancer Society. They gave him a small grant and using that funding, Dr. Prasher was able to clone GFP. The GFP gene. So essentially he was able to make copies of the gene and able to isolate almost the complete DNA sequence. Today that would be I think like figure one A of a paper, maybe, but... Jyoti Asundi (47:05) Or even or even in the supplemental, yeah, correct. But that was that was the groundbreaking. Aarati Asundi (47:08) Yeah, but back then... yeah, that was groundbreaking. That was huge with the technology limitations that they had, huge achievement. Jyoti Asundi (47:17) Even to actually sequester out the DNA. The techniques were so bad, it was all radioactive based in those days. Aarati Asundi (47:24) So Dr. Prasher was able to write a paper on this and also suggested in that paper that GFP could be used as a marker for other molecules, but before he could take his work much further, his funding ran out. Around this time, Roger read this paper and reached out to Dr. Prasher, as did another scientist, Martin Chalfie. Martin Chalfie was interested in putting GFP in different animals, and he was successful in expressing GFP in E. coli bacteria and in C. elegans, my grad school model organism. Jyoti Asundi (48:02) Yes. Your grad school work. Yes. Aarati Asundi (48:05) Yes, my little worm. And so Martin Chaflie was able to show that GFP could in fact work as a fluorescent tag in living animals. Roger, on the other hand, was more interested in expanding the toolkit. Remember, his work with protein kinase and cyclic AMP had required two colors. He had the green fluorescein and the red rhodamine dye. Jyoti Asundi (48:29) So now he needs a red guy. Aarati Asundi (48:32) Yeah, so GFP gives us green, and Roger's like, let's make more colors. In order to do that, he first had to understand how GFP worked in the first place. A lot of the other bioluminescent proteins that they had found in nature from other organisms usually required some other enzyme or cofactor to actually fluoresce. Jyoti Asundi (48:56) Okay. Aarati Asundi (48:57) And that's what kind of made them bad for science, because then you needed to have like two things that you're putting into this cell. Jyoti Asundi (49:00) Okay. Yet another- yet another component. Yeah, it becomes -the system becomes more and more complex. You're add already adding these fluorescent proteins to understand a complex system with various moving parts. And you're hoping just by adding one tag you can decode it. But now in order to decode the tag, you need another decoder. So it becomes even more complex. So you need to keep at least the fairly simple. Aarati Asundi (49:29) Yes. But Roger found that GFP was special because it worked all on its own as long as oxygen was present. If oxygen was there, you could get this oxidation reaction that would make the GFP glow. Jyoti Asundi (49:44) Okay. Aarati Asundi (49:44) He also looked at the crystal structure of GFP and found that when it folded, it formed a special structure that almost looks like a cylindrical lantern with a glowing light on the inside. That glowing part on the inside is called a chromophore and it absorbs blue light and emits green light. Jyoti Asundi (50:06) Yes. Aarati Asundi (50:07) And then the rest of the protein structure of GFP that forms the cylindrical lantern protects that light from water or any other unwanted chemical reactions. Jyoti Asundi (50:17) So pretty! Aarati Asundi (50:18) Yes, it's really cool. And so now because he had spent so much time understanding the structure of GFP, he and his lab could start to make small modifications to the DNA sequence and start to make alternate forms of GFP that either fluoresced at different wavelengths. So for example, they were able to make YFP, yellow fluorescent protein... Jyoti Asundi (50:43) Yes, yes. Aarati Asundi (50:44) BFP, so blue. Or CFP cyan. So they were just able to shift the green wavelength yeah. Jyoti Asundi (50:52) The wavelengths at yeah, the activation and the emission wavelengths have changed a bit. ⁓ Aarati Asundi (50:58) Yeah, so he was able to shift it a little bit left or right to make it yellow or blue And they were also able to create brighter versions of GFP that gave a stronger fluorescent signal than the wild type. Roger and his lab tried for some time to get a red fluorescent protein, but they couldn't figure out how to do it. They couldn't shift the, emitted wavelength that far away from green. That is until another scientist, Dr. Sergei Lukyanov's lab at the Russian Academy of Science got involved. They discovered a species of red coral called discosoma that got its red color from a homologue of GFP. Jyoti Asundi (51:41) Another Brother. Aarati Asundi (51:43) Yeah, and no one had looked at the coral before because the coral itself wasn't bioluminescent. But it turned out that the protein that makes it red is a fluorescent protein. And so finally now Roger and his lab were basically able to like look at nature's cheat code and see like how did nature do it? And it turns out the red fluorescent protein, which was called DS red, DS for discosoma, it had a chromophore that looked exactly like GFP, but with an extra structure attached. And they were now able to look at that s structure and study that structure and see... Jyoti Asundi (52:19) Yeah and now you add that to GFP and make.. got it. Aarati Asundi (52:23) And make it shift to red. Jyoti Asundi (52:25) Amazing. Nature... the lesson stays the same. Aarati Asundi (52:28) Can't beat it. Jyoti Asundi (52:29) Can't beat nature. Learn from nature, observe nature. Aarati Asundi (52:33) And that's actually what Roger was saying too. He was like, none of this would have been possible without the jellyfish. People are all saying, you know, my gosh, you're so amazing. You won the Nobel Prize. And he's like, "The jellyfish is the one who did it." Jyoti Asundi (52:46) Yeah, it's the jellyfish who is amazing. Nature that's amazing. I just borrowed her cheat sheet. Aarati Asundi (52:52) Yeah, exactly. So now that they've figured out DSred, again they're able to start making mutations and they start making slightly different reds. They got like a more orange-ish red, a deeper maroon red, a scarlet, all these various shades of red. But what can you name them all? You can't name them all RFP. They're all slightly different. And so that's when they started getting creative with the names. And that's how we get the names mCherry, mStrawberry, mTomato, and mRaspberry. There's like- they have all different fruits. Jyoti Asundi (53:28) Yes. I have worked with so many of these and it's only now that I know how the names derived because until now I have only focused on what my needs are and how different I need the colors to be and what are the filters- excitation and emission filters- that I have available to me for my experiment and then choose the appropriate color based on that. Aarati Asundi (53:52) So he created this whole rainbow of colors, basically. Jyoti Asundi (53:56) Nice. Aarati Asundi (53:57) And by the way, if you're why it's "m" Cherry and "m" Raspberry, the "m" stands for monomeric. The original DSred in the coral formed tetramers, which would have messed up our experiments and so Roger's lab did a lot of work getting the proteins to work as monomers. Jyoti Asundi (54:15) That's a lot of work they have put in. So many steps. Aarati Asundi (54:17) Yes, yes. Oh my gosh, I am skating over so much of his work. I am skating over so much. Jyoti Asundi (54:20) You're just saying it all. Yeah, no even every step that he's taking, I can see the amount of effort. Like multiple people have to spend years making it happen. We are just able to say it like click, click, click one sentence here and two sentences there. But no, this is intense failure and no, that didn't work, and let's try this and make this happen change this and change this and until you finally get where you are. And the perseverance that is required for that... Aarati Asundi (54:51) Yeah, and I was listening to Nobel Prize speech, and in that, when he like he's describing his work, it's just incredible the amount of work and the depth of knowledge he has. He was like, we mutated this serine to a threonine and then we- that didn't work, and so we then went back and we're like, but the serine is important, so why don't we try making the glycine instead? And he's like got the whole amino acid sequence down, the whole DNA sequence down. He knows exactly how things fold and which amino acids interact with each other. And if you change that one, you can't change it. Otherwise it screws up everything. But you could do whatever you want to that other one. So we were messing around with that other one. And I was just like, my God, this is just, you know... Jyoti Asundi (55:30) Wow, just the ultimate master who knows exactly how to move his little chess pieces. Beautiful, beautiful. Aarati Asundi (55:39) Yes, yes. So really amazing. I'm skating over so much. I'm like, you know, just summarizing so much work. Jyoti Asundi (55:46) Yes, otherwise we'd be here for the next three episodes just talking about this person. Aarati Asundi (55:50) Yeah, for real. And so finally he's like achieved his dream. He's got red, he's got yellow, green, blue, and now scientists could use all of these colors to make these beautiful scientific really enlightening images that show us very precisely different structures, which proteins are interacting with each other, where cells are located, all these sorts of things. In 2008, Roger got woken up by a call in the middle of the night from the Royal Swedish Academy saying that he had won the Nobel Prize for his work on fluorescent proteins. Jyoti Asundi (56:29) Wow. Aarati Asundi (56:30) He shared the prize with Dr. Osamu Shimamura... Jyoti Asundi (56:34) The one who actually found the GFP to begin with. Aarati Asundi (56:38) Yes, all the way back in the sixties. And then Dr. Martin Chalfie, who was the one who tagged it in bacteria and C. elegans. Who used GFP to tag proteins. Yeah. Jyoti Asundi (56:53) Oh okay, okay. So the one who actually isolated it, Prasher, didn't get included in this? Aarati Asundi (57:01) Correct. And so that was actually a bit of a controversy for this Nobel Prize. He was the one who cloned GFP and was left out. The prize can only be shared by up to three people. And after his funding had run out, after he had cloned GFP Douglas Prasher had struggled to find a job in science and he actually ran out of his savings and he ended up leaving science altogether and becoming a shuttle bus driver for a Toyota dealership. So that's what he was doing when he heard about the Nobel Prize being awarded in GFP. Jyoti Asundi (57:41) Oh that's heartbreaking. I'm heartbroken. Aarati Asundi (57:45) He said he was very happy for the winners, although he was somewhat surprised that the fluorescent protein from a jellyfish ended up being that important that it went on to win the Nobel Prize. All three of the Nobel Prize winners thanked Douglas Prasher profusely in their Nobel Prize speeches, saying that his work had been invaluable. And Martin Chafie even said they really should have given his spot to Douglas Prasher and just left him out of it entirely. Jyoti Asundi (58:15) Actually, that is my sentiment as well, although I am not the right person to talk about these kind of things. I'm too- I'm speaking too big for my boots. But ⁓ yeah, I think Prasher... Aarati Asundi (58:24) Yeah, like I don't know how they made these decisions, but yeah. Jyoti Asundi (58:28) Correct. Yeah. I think the fact that he had to give up science played a role there. The committee might have found it "Oh he's divorced from science now, so let's leave him alone" or kind of thing. That's not fair. Aarati Asundi (58:42) Yeah, maybe. It might have been. Jyoti Asundi (58:43) Very difficult. Very difficult. Aarati Asundi (58:45) I don't know. Roger invited Douglas Prasher to the Nobel ceremony as his personal guest and paid for all of his expenses to travel there and stay there. Roger also invited Douglas to join his lab at UC San Diego and Douglas finally accepted that offer in 2012. Jyoti Asundi (59:03) Okay, scientists looking out for each other to the best of their ability. Aarati Asundi (59:07) Trying, yeah. Roger continued developing fluorescent proteins, even making an infrared fluorescent protein. He started collaborating with medical scientists to find uses for his proteins, like using them to light up cancer cells so that surgeons could more easily remove tumors. Jyoti Asundi (59:29) Yes. Aarati Asundi (59:30) And using them to understand long term memory storage in the brain. He also found that fluorescent proteins, surprise, surprise, were a great way to get high schoolers interested in science because they got to play with pretty bright colors. Jyoti Asundi (59:42) Yes, who doesn't like to play with colors? Que young Roger who played with the potassium permanganate. Aarati Asundi (59:50) Mm-hmm. Exactly. That's what got him into science in the first place, you know, like seeing all the beautiful colors, watching them. Yeah and so things really came full circle. Roger started getting involved with outreach programs and student lunches students who are interested in science. Sadly, in 2016, Roger died suddenly while biking on a trail in Eugene, Oregon. By this time, he was a part-time lecturer at UCSD, but he was spending time doing a sabbatical at the University of Oregon. Jyoti Asundi (1:00:29) Okay. Aarati Asundi (1:00:30) He was only 64, and so his death was very unexpected. Although his family did not disclose the exact cause of death, it may be important to note that he had survived a stroke and a battle with prostate cancer a few years earlier. Jyoti Asundi (1:00:45) Okay, so there had been some problems beforehand. Okay. Aarati Asundi (1:00:50) Yes. And the fact that he died while, biking, I'm assuming a strenuous trail, you know, it sounded like he was maybe pushing himself really hard that day or something, and you know... Jyoti Asundi (1:01:04) Could be. Yeah. Aarati Asundi (1:01:04) Maybe something gave out. Not sure, regardless, he was taken from us too soon. But there's no doubt that he absolutely changed, the scientific world. And if you go to Wikipedia and you look at the number, the sheer number of awards that he got, starting all the way back in high school with him winning that $10,000 scholarship prize, you know, throughout his life, it's just like this huge list of awards that he's won. And yeah, that's the story Roger Tsien. Jyoti Asundi (1:01:40) Amazing. And before we forget we should take a sip in his name as well and for the all the three scientists who actually won the prize. Aarati Asundi (1:01:43) Yes. All the three scientists. Yes. Salud. Jyoti Asundi (1:01:52) More power to science. We need more people like Roger, and we need more people like Prasher also. Aarati Asundi (1:02:00) Yeah, some of these people are just really amazing and really big people, you know, like it doesn't seem like he harbored any bitterness really at all, or not that he showed to outside world. He was always just very happy, you know, when Martin Chalfie and Roger Tsien contacted him about his 1992 paper saying, "Hey, we read your paper, we wanna study GFP some more." He gave them everything. He was like, "Here's all the information I have, here's some samples that I created, here's, you know, some protein that I've isolated. Take it all." Jyoti Asundi (1:02:35) So generous. Aarati Asundi (1:02:36) Yeah do everything, you know, do what you need to do. Jyoti Asundi (1:02:39) So Chalfie and Roger basically built their work on his shoulders... Aarati Asundi (1:02:44) Mm-hmm. Yes. Jyoti Asundi (1:02:45) ...and yet he didn't get recognized. Aarati Asundi (1:02:48) Yeah. And they were fully cognizant of that. Both Roger and Martin Chalfie were like, you know, he really deserves to be up here with us. Jyoti Asundi (1:02:56) Yes. Wow. Amazing, amazing story. Thank you for sharing. Aarati Asundi (1:03:02) That you probably never knew about GFP, right? Like who knew all of this history behind it? Jyoti Asundi (1:03:05) I did not know. I did not know this much information. Like I said, I'm so focused on how to make this work that I don't really often have time to deep dive into "Who might have thought of making this happen?" Aarati Asundi (1:03:22) Pretty amazing story I thought. So yeah. Jyoti Asundi (1:03:24) Yes. Awesome. Thank you so much. Beautiful story. Aarati Asundi (1:03:28) Thanks for listening. If you have a suggestion for a story we should cover or thoughts you want to share about an episode, reach out to us at smartteapodcast.com. You can follow us on Instagram, TikTok, and Blue Sky @smartteapodcast and listen to us on Spotify, Apple Podcasts, YouTube, or wherever you get your podcasts. And leave us a rating or comment. It helps us grow. New episodes are released every other Wednesday. See you next time!