 try to develop 3D flights and imaging techniques in order to visualize the biological process better with higher spatial resolution and also be able to imagine longer and faster to capture the details of how biology happened at protein level or cellular level or multicellular level. That information is a fortune. I think it can lead us to a complete understanding in bioscience and in related to our health and for clinical applications, disease diagnostic etc because it's tissue right and then that means you can correlate you can connect this to billion dollar even trillion dollar business and that is the impact I think all the future I'm looking for. That's why we are giving our entire design software everything to research for non-commercial use. We are happy very happy actually we are honored to do that to return our knowledge to the society not only to to China I mean to to everyone in the world. I know someone's opinion may contradict yours. Where's my friend Alan? It's all about your perspective. Who are we and what is the nature of this reality? Five, four, three, two, one. What's up everyone? Welcome to Simulation. I'm your host Alan Sakyan. We are on site at the beautiful Westlake University in Hangzhou, China. We are now going to be talking about Tiling Lightsheet Microscopy. We have Dr. Liang. Hi everyone. Hi Liang. Thank you so much for joining us on the show. Thank you for having me. Yes. Really appreciate you coming on. Thanks. Thank you very much for those who don't know Liang's background. He's an associate professor at Westlake University pioneering new optical methods enabling rapid high resolution 3d imaging of expanded biological specimens and you can find the links in the bio below. Liang let's start things off with one of our favorite questions we like asking our guests. What are your thoughts on the direction of our world? Well that's a big question. I think definitely we are trying to understand our life. We are trying to understand each other better and we are trying to build a better world for everybody, for ourselves. I think that's a goal and the direction of the world. Of course there are a lot of back and forth possibly but in the end I think we are going to reach there. Basically we are going to reach a place that is that work better for everyone and for everybody hopefully can get their right destiny. Yes. And what do you think is an important skill for us to embody to ensure that we continuously improve our civilization? I think one of the most important things is perhaps again let's say so basically of course in order to achieve something you have to basically be regulated and be regulated means too regulated on two part of our life. One is regulation on others the second is regulation ourselves. So basically we perhaps cannot put a lot of hope in how to regulate other people's behavior but what we can put our efforts in or what we can do better certainly is we can regulate ourselves better and if we truly understand the meaning of life understand what we are where we are marching on and what's the goal of our life I think we can regulate ourselves better and if meanwhile if they have better understanding better communication make sure that everybody understand each other and in the end I think we can reach that goal by better regulation of ourselves. Yes perhaps yeah if we think about what's the most important part perhaps that's a very important thing yes. Yeah yeah that's definitely in the top some of the oldest advice know thyself and also do really good job at regulating your emotions your goals your aspirations your family relationships your career the community around you this is very very important critical like that a lot yeah focus regulation of the self. Exactly. I love that. Let's talk about you as a child so let's talk about your journey where were you born how did you get interested in optics and science. Okay so I was born in Shanxi province in China and at one year old my parents moved to most western part of China because of their job the province called Xinjiang province of China so with a lot of minority people and then I moved there at one and then I grew up there until I was 17th after that I was admitted into Qinghua University in Beijing and then I went there for college I spent seven years in that university my major was called it's called precision instrumentation so I built I learned how to build instrument I got my bachelor and master degree in that university so it's a good university at some point let's say in most people's mind it's a very good university I do agree with but there are something I think I didn't learn during that period time I was trained in a lot of how to optimize a lot of details I was training electronics mechanics everything but in the end I don't know what kind of instrument I want to build I have very little understanding on instrument industry because it was not so let's say developed in China at that period of time in maybe year of 2000 and then I was thinking about why I'm doing this and then I searched around I searched what kind of areas need more instrument and at that point in the time I learned actually analytical instrumentation is a very let's say important industrial area and they do need all kinds of instrument and to analyze the chemicals we are dealing with every day and one of the techniques called mass spectrometry so basically it's analyzed molecular weight and by doing that we can know what chemicals are there in the samples and then while I thought that's a great direction I mean while my wife went to the US and she went to the Purdue University and fortunately there was a very famous professor called Graham Cooks in the chemistry department of Purdue University and he was an expert in mass spectrometry especially instrumentation mass spectrometry basically how to build better mass spectrometer and my major was precision instrument so it's a perfect match and then I think while I can go to that department and learn what's a real instrument that people need and I can maybe find a better goal of my life learn what I want to do in the rest of my life to reach a better career right so I also applied that university and indeed joined Graham Cooks lab and doing mass spectrometry during my PhD that was from 2005 to 2009 until I graduated so indeed during that period time I built let's say the first class let's say the instrument I built called miniature mass spectrometer the reason that direction is important is because the conventional mass spectrometer are huge very large mean usually typically weights about 100 kilogram or even more meanwhile there are a lot of applications you want a very quick measurement fast fast response measurement of the chemical environment for example in the industry side for example another important application is in astronomy so we send the satellite to different for example to Mars right but we do want to know what's the chemicals on Mars so the best one of the best way actually indeed send mass spectrometers to Mars to measure the chemical components that planet and then of course you need to build a very small but precise mass spectrometer in order to do that and so that's a major motivation of my research during that period time and I indeed build very small mass spectrometers one of them is only five kilogram and it can do a lot of awesome things and actually I invented a very important device called the discontinuous atmospheric pressure interface to introduce ions from atmosphere to vacuum space efficiently with a very small equipment and actually that device was used on a on the latest mass rover built by I think European what's that EPS so basically it's X for Mars 2020 it's going to be launched in yeah next year and then yes that device was actually used in the latest mass rover going to be sent to Mars but anyway so that was what I was doing during my PhD I think it was pretty successful I learned many things in that lab and then of course in 2009 I graduated from Purdue right let's let's stay on the subject for a little bit longer than sure yes so miniaturizing mass spectrometry yes so taking the machine size from 100 kilograms right and bringing it down to five exactly so it can do things like go on satellites to Mars exactly because it's expensive to launch things into space exactly yeah and so that's one of the relatable examples and then also just this move in general so your your wife was first right first to produce yes exactly following okay cool cool and then what was it like when you first arrived and you were immersing in the United States culture what was that like and also what was it like when you discovered that you could miniaturize some a machine that was 100 kilograms down to five how did you come up with those ideas right so let's first let's talk about what's my feeling when I first went to the US so basically I until I went moment I went to the United States I haven't been traveling to any other country so the entire world is quite that's a very fresh very mystery place to me I so I was very excited to be basically everything happening there when I arrived is very attractive very new to me and I think I adapted to the environment quickly because I think let's say one of the most impressive things I think everybody was so nice gentle and plight that's perhaps one reason I could adapt into the environment quickly and also the other thing I realized is I had a lot of freedom to do what I want although I was paid I was supported by Purdue University by Dr. Graham Cooks with research assistantship so I need to do research in Islam but actually I was not required to do some specific thing I do have the space to think about my own ideas to find the topic that I think interesting and what I need to do so that's something I most impressive to me so basically the freedom and how nice how supportive the environment was when I arrived in the US and also the other thing is I think because at my period of time even I think even by now when we think about the most advanced technology we always think about the United States right so at that point also when I was at Tsinghua I think we did some pretty awesome project but one thing we are not sure is we don't have the we didn't have the confidence about the level of our project was we don't know what's the what the world world first class project what's the best project what's the frontier really frontier problems people are working on and we have no clue of that but after I went to the United States and joined the Dr. Graham Cooks lab because he was a leader in that area I knew wow this is the the the most frontier project that people are working on and this is the cutting edge technology and then I have the confidence of what I'm working on represents the highest level of this field I think that's very important for the development of young scientists basically establish the confidence of your own ability your own vision your own judgment I think that's something I gained I mean from the experiences in Purdue and after that I mean because you know you are working on the most let's say frontier project and you do can release your your mind right you can think about whatever crazy ideas to work on instead of suspecting whether this is something makes sense at all right because you know you know the status of the frontier so anything that is new into your mind you know it is new for the entire field right and you can just move forward with that idea I think that's something um very important to me and I think to anyone who was trained in the United States yes know where the frontier is and then have the confidence in the vision to do the work at the frontier so then you guys how did you have the foresight to say okay how do we bring this down the mass spectrometry down in right so basically that is project not started by myself so basically it it has a history actually this project has a history go back to maybe 60s there was a scientist in University of Minnesota I'm sorry I cannot remember his full name his Italian American I think his last name of person is Alfredo so he basically he was a part of the Manhattan project right that nuclear project basically used mass battery geometry to separate uranium to make it more to purify the the chemical and then at some point in the 1960s 70s the United States started that aerospace project of course to and then people had the need to send the mass analyzer into the space and he started that idea say if we can build miniature mass spectrometers and then we can actually certain certain goals and indeed people start to build a different version of mass spectrometers and make them small and then send them to space to actually many different planets but of course the techniques is developing right so we can build better and better instrument with more analytical ability to analyze more complicated chemicals for example at the beginning people will be happy to see whether there is water whether it's there oxygen or carbon dioxide what's the concentration of them in the atmosphere right but at some point people would like to see whether there's amino acid whether there could be protein or more complicated molecules on those planets so we can learn more about the environment of those planets right and then so by inherit the previous knowledge and also on the other hand because I was trained in instrumentation so although I was in a chemistry department a lot of my colleagues or many others still they were chemists but I'm actually a mechanist or let's say instrument let's say how does that work so I build instrument I know how to make those things happen so it's it's just so natural to me and actually it's a yeah that's why I think I suddenly find a place that I can apply what I learned into the into some very important and interesting project so yeah things happen so natural to me that's my feeling another thing I think I have to appreciate is Dr. Graham Cooks has a lot of collaboration with industrial companies for example the Somo Fisher now called Somo Fisher at that point there was a mass spec company called Finnegan Somo Finnegan they build let's say the best mass spectrometers in the world and they have the biggest market share at this point so they do have a lot of collaborations and then from that collaboration I think I also learned the organization of big companies how they are starting a project why how do they make decision about whether they want to start initiate a product or not I mean what going to be kind of resources going you are going to need in order to make that happen kind of thing so indeed yes so basically part of my training happened naturally about my research about my instrumentation ideas etc many of them are just I think they emerge gradually during my education why I learn more and more stuff during those years and and also when you are in an environment that you are encouraged to think actively because I do have the freedom right because he my advisor was so busy he barely had time to talk to me so when you are in that environment you can sing freely and you have the resource and indeed I mean it's um yeah they're not much better environment than that to really produce some good research results yes yeah thinking freely having the resources and also those collaborative partnerships that you were talking about exactly very crucial let's go to the postdoc then sure so this was at Howard Hughes Medical Institute yes and so what were you doing there so I was building a fluorescence microscope there and try to basically try to develop a 3d fluorescent imaging technique in order to visualize the biological process better with higher spatial resolution and also be able to imagine longer and faster to capture the details of how biology happened at protein level or cellular level or multicellular level yeah actually this is also an interesting transition because mass spectrometry has nothing to do with fluorescent microscopy right that's another thing actually an important decision in my career so the reason that after I graduated from Purdue I mean let's say I learned several things I mean why why you make a decision to do certain things to do certain things to decide what is your career I think you have to find something you enjoy right if you don't enjoy it you cannot do it well you have you have to find something that you think it's important you can convince yourself to spend a lot of time on that thing right and then you have to think of in what way you can make impact right and miniature mass spectrometry or miniature mass spectrometer it's a very cool project but at that point I don't see how it can influence a lot of people because yes sense something to Mars is very important but only very small amount of people going to care that thing right I mean versus the benefits of being able to unlock the biology of exactly so so I think I indeed I want to choose more important projects so basically everybody wants to have more positive impact on the society on the environment right so I decide to change my research direction let's say the other reason I decide to quit from mass spectrometry field is because of mass spectrometry is a very complicated instrument and it's it's very well established field because it's it has hundreds of billion dollar market that's why there are a lot of big companies like angel and like thermal waters they spend a lot of resources in their own r&d and for an academy lab in order to compete with them to build a better instrument it's almost a mission impossible because they have very experienced for experienced engineers they have people work on a small trivial small project but very important thing for many many years to improve the performance right there's no way for a research lab to compete with industry company in that level basically in any well-defined routine of science or any technology I mean academics just cannot compete with companies that's something I learned because they have resources and momentum to optimize things better right and I learned I have to pick something that important has big impact but but not that attractive to to those big companies and I thought life science is perhaps a field I should step into and imaging is something seems so interesting because I was always amazed by those images and then yes that's why I made the decision to get into flies and microscopy and then the reason is where you want to do your post out right because in this state if you know let's say if your PhD you want to do something completely different from your PhD during a post out you are having very big risks first nobody going to want you because why right because you cannot do anything second the second thing is you are risking yourself your own career right and then although I know actually but of course the important thing is you want to convince people because for myself is if I have interest in something if I can convince myself to do something I will spend my time I would do my best and after I graduated from Purdue I think I picked up that confidence that I can if I want to handle it well I want to do it well I could do it that's some I was pretty confident at the point of time and you started high neuroplasticity too to be able to make this right right and then I checked all those labs about which lab are doing let's say frontier imaging what's a frontier image labs and then so occasionally I read a report about every basic so yeah if you had a chance you should check his story very interesting very so he's such an inspiring person that he has he had actually a very difficult career in his I think I think it's a difficult successful but difficult because he changed his career many times he used to be a scientist and then used and then became a mechanical engineer in his father's factory self-employed or jobless let's say and then come back to academia and become a win the world prize what inspired me is that this of course in 2009 hitting a Nobel prize but he published a very important paper about super evolution the paper actually get him a Nobel prize and then I saw an increased resolution with my cross yeah break the diffraction limit and then the diffraction yeah so basically uh right usually we can our resolution resolution of flights and the imaging is dominated by the fraction limit of light right we can see maybe 200 nanometers that's the best we can see but with his method we can drive that resolution down to about 10 nanometer resolution that's a reason so that's his idea and he indeed actually practiced that idea make that made it happen that's why he won the prize and he did a lot of other important um uh researches in his career as well and explain this to us um why is there a diffraction limit with light oh so basically the light is actually an electrical magnetic wave right and then if it's a wave then it means it has certain wavelengths and then in order to create image basically we need to use that wave to create a focus right basically we need to collect a lot of light and then focus them into a spot because that light has a certain wavelength it won't be past the the the minimum spot size you can create with that light cannot be less than half of the wavelengths of that light and then that is called the diffraction limit for example if we use green light to do image and then the wavelengths of the green light is perhaps somewhere around 520 nanometer and then we divide that by two is about 260 nanometer of course if we can if we use different image buffer we can push that resolution a little bit lower to maybe 200 kind of thing but that's best you can do without doing anything else but with his method you are able to push the resolution down to 10 nanometers and is that going further on the electromagnetic spectrum past the past the violet is that are we going how how do you get down past 200 nanometers that method was very clever it's actually so basically there is a requirement let's say let's start a little bit make a story a little bit longer basically how we can see a stuff let's say see a cell see its structure single yeah they don't emit light right so there is no way we can see them by any let's say equipment or by our bare eyes without seeing any without doing anything to the sample we have to shine light added and images yeah and then the more important thing is we have to add a probe add something that can emit light onto the object that we want to study so what people did was another Nobel Prize work actually it's called GFP it's Roger Chen won that prize and a few other scientists so basically you label a molecular probe a protein probe genetically to the molecule you want to study and that probe is called GFP so basically when you shine a laser to that molecule and that molecule going to emit fluorescence and by clad that fluorescence you will be able to see that molecule right but if you just do that regular observation you won't be able to exceed the diffraction limit unless one thing you can do is if those labels if they emit one by one not emit at the same time so basically what are the images you are going to see is just a random spot right however if you know that your molecules are emitting light one by one and you see a random spread a spot you know that that spot the center of spot is a molecule right and then you can use that image to localize the position of that molecule and you repeat this process to let all molecules shine emit in sequence and then you can find the location of all your forensic probes right and in the end you create an image that passes a diffraction limit of light so basically this is Eric's idea I'm trying to make a specifics part of the cell like a protein fluoresce so I can image it right and that is already past the diffraction that is not past the diffraction limit but the way you collect the signal and find the location of the molecule is a key is past the diffraction exactly okay because we're a protein in a cell is about how many nanometers the protein of a cell about four to five nanometer as well that's a typical size of a of a protein they're larger ones smaller ones yeah but a few nanometers typically yes so the first your first like aha moments were that you can do a microscopy at even sub 200 nanometer levels and you wanted to go even deeper into understanding how to do that right build new tools yeah to enable us to go deeper than right actually a little different direction though but anyway that was Eric's work right and that was what inspired me to join his lab because I knew wow this is such a smart person and it is so cool and I although I knew nothing about my crossbeam because he was not that famous as a pure time I knew I had a chance to join his lab and I knew this is must be a person I like because he has been following his dream in his career and I like this idea so I think we can perhaps we can have some echo each other right and then I write a long email to him and he invited me to to interview and then I joined his lab and then I started to work on 3d flights and imaging which is actually another dimension of imaging let's think about this way if you want to understand the life right you need to understand two things you need two kind of information one is a spatial resolution right basically you have to see the details or you have to see the object and the other dimension is you want to see the process you want to see things alive right because you feel for any super resolution technique almost super resolution technique what do you have is a very high resolution image of a dead sample of a fixed sample a dead stuff at a moment on a plane that's the most of time you can get me how much you can interpret this life a living process from that information not enough right and then you will need you will want to have a image alive dynamic 3d process alive with high resolution and that's a different challenge and that is a project I was working on when I was in Eric's lab and since that actually so the differences between like something as a very thin slice of some tissue over one moment in time versus the already dead versus something that's alive yeah like a drosophila fruit right and being able to image while it's alive over time right the brain or something right you can think about that way or you can say you let's say you are having a hd image let's say 4k image in your hands about a football game or you prefer to say to let's say to watch the game at maybe 480 pixel resolution for one play just one play versus the whole game exactly so basically actually you want both right you want you want the high resolution live video of the entire process but at that point of time 2010 there were some several good live imaging technique but I like that sports analogy a lot right a really good one actually sure one seeing one play of the game and trying to make some inference about how the biology works versus watching the whole game in high definition exactly that's a really good yeah so in the end we are collecting information of the process live process right so basically and then that was the that's in the motivation of my project hard started and of course at beginning when I joined his lab I mean Eric gave me the idea called best of being playing information my cross p I mean I had so at that time I have no idea whether that's a good idea bad idea working on not working idea but because I have trust on him so I trust his idea and I just I think I will be successful just following his instruction and make it happen I think I will learn things so in the end I worked very hard at that period of time that's another advice perhaps to most people or young younger generations so when I joined his lab Eric asked me in his email he asked him he just emailed me said can you work 80 hours per week if you cannot just don't come right you cannot have anything down if you can yeah you are welcome yes and I think to be honest when you work on something like 80 hours a week I mean I'm willing to do that I mean because if I am I was not working for 80 hours per week I said I'm actually wasting my time because there's a reason I go there is I want to learn this stuff right I want to learn as quick as possible and I want to learn from the best scientists so I won't waste my time so I push the front here exactly what would you be doing yeah otherwise what how otherwise home and watch TV instead yeah exactly then nothing more interesting than the work at some point right yeah so yeah not a problem at all and I joined his lab and worked on that project in indeed that was a very fantastic project I think we we were the first group showing very indeed I mean subcellular high resolution 3d light process of cell behaviors subcellular behaviors etc so that was in 2011 we published our first paper about best of implementation microscopy on nature methods with my colleagues so basically with arab basic and another colleague called Thomas planchon he is a associate professor in Delaware University now state university so yeah so and at that time our group just have three people so all of us worked on project we indeed pushed the limit of that technique and we generally read some very cool stuff I think we initially that research direction of of improve the 3d image ability of lychee microscopy by optimizing the lychee profile and the technique we use called best of being plain nature microscopy and also I think we were the first group showing how fantastic it is if you can indeed watch the live biological process with high spatial and temporal resolution and after that me I think this field become enormously attractive and popular because people realize that well just by spatial resolution improvement you cannot understand too much you have to get it alive and you have to do something in order to understand the entire process right that's pretty much what I was doing during my post-op research with arab basic yeah and now take us to when you're doing your system professorship at SUNY stony brook right and you're doing tiling twin lattice light sheets yeah more accurately it's called tiling light sheet microscopy right tiling light sheet yeah so that's a technique I invented when I was a system professor at the stony brook University the reason my research migrated to that technique is because um so indeed we developed some awesome technique right 3d live unit technique when I was in Eric's business lab however that limit that techniques also technique also had had a lot of limitations so at the beginning we we thought we want to develop a lychee technique that can image large specimens at a high spatial resolution with high image speed so we can watch the dynamics of of biologic specimens right for example we want to study how embryos developed we want to study how what's a neural activities in in a brain of liverfish etc so and then we indeed we were pretty successful developed some technique and applied them to cellular specimens for example we can image a cultured cell on car sleep very well we can image that dynamic process very well but it was very difficult to apply the same technique to larger specimens for example liverfish embryos or cialic embryos some some models that biologists use very often to study biology question right so basically you cannot get the same result on multi-cell specimens uh as you what you can get on single cell specimens and I was confused by why that and I didn't get that answer when I was doing postdoc in Eric's lab the other question I was trying to answer was um at beginning you would be pretty happy when you first seen those dynamic process those videos right I mean because it's just quite fantastic to see those live in process but at some point you would like to understand why right because after seeing what's happening you all the next question is always why these things are doing those are doing that why what are they doing what are those molecules are doing why the cell migrates from A and B and what reason cause that result right you always want to answer this question and you will find that even you collect those information if you don't analyze them properly you won't be able to use this information you have to interpret the information in certain way in order to reach a conclusion right that's the second question I think we didn't answer during my postal research and then I started to work on this um issues when I was at Sune Brook so that's why how I started to work on 3d imaging of large multi-cell specimens and how I started to work on quantitative understanding of cellular behaviors in multi-cell environments the reason that is difficult let's say the difficulties from several directions first you want to study cells in their native environment right for example a embryo embryo has a lot of cells right it's a very complete environment it's like it's it's you can consider like a smaller society right community right people are interact with each other and people are doing their own stuff but people are communicating right as a behavior of individual cell or individual person is often result from other people inside your own decision right it's the same thing happening in a multi-cell specimen and you have to gain that detail you have to have tools to be able to collect those information in order to understand them right that's the first the second is you have to have a method to understand to organize the information and reach your conclusion you have to have a model you have to have a certain way that's an accurate definition of those behaviors and you have to have a method and to analyze those behavior to find the relationship between them and then that's a difficult or challenging and none of this exists at that point of time so that's the the motivation behind my research and then of course the question is how are you going to collect that information that's a high spatial and temporal resolution from large specimens right and let's say one thing we learned in my post-doc research is one way use our technique called lysine microscopy to image a 3d behavior we want to a key issue is you want to create a large and thin lysine in order to image a 3d process well because you have to scan that lysine across your sample but after five years of research we finally realized there's no way you can create a large and thin lysine at the same time basically means if we proceed along the direction we have been during my post-doc research we won't be able to actually this go so this looks like a piece of tissue that you're imaging and your shining laser right it threw it in a very thin sheet right and then that's lighting it up in the exact image right so basically imagine you have a tissue right i want to see see the layer by layer right what i need to do is i can shine a very thin layer of light to a selective plane of that tissue and i scan the layer across the sample i capture a lot of images and i get a high resolution image of that sample right but how you can create a thin layer of light is is a is an important question but in the end we realized no you cannot create a thin layer of light over a long distance so my technique called tiling lysine microscopy was yes we cannot create a long and thin layer of light but we can create a we can create a short but thin layer of light and then what we are doing is we scan the move that layer of light quickly within the image plan and then by you capture multiple images right by doing that your it's equivalent to a let's virtual large and thin layer sheet and by using this strategy you are able to image large multi-cellular specimens with high resolution so you're you're you're modulating this this laser light from a point to another point to another point exactly and then imaging as you modulate it to those different spots exactly so basically that's another thing let's say the compromise of life basically at beginning we are looking for something perfect right we want high resolution we want high image speed we want to image things for a very long period of time and with high signal signal to noise ratio but at some point you will realize it's impossible it's it's just like everything right it's sometimes very difficult when you ask for everything but if you can think about exactly what's most important thing for a specific moment or for some specific experiment you always find that you can sacrifice something so in tiny life sheet microscopy what you are sacrificing because you have to move light in the field of view right you sacrifice your your your image speed a little bit but because the technique is already fast you can you still can maintain a reasonable speed to to finish that image process and because if you don't do that compromise you cannot see it you there you get nothing but with this technique yes you compromise something but you gain something at the same time and another advantage of this technique is that you can you can basically you have very large freedom to decide I mean how sick that light sheet is how fast you want to tell it so you can have great flexibility to optimize the experiment and then by doing that you have more chance to get the information you want from a biological process and thereafter you can start to create a model and you can analyze your 3d images and start to understand the relationship between them that's another reason actually why 3d imaging is important for quantitative analysis because if you don't capture an entire process in 3d if you only capture a certain plan yes you can guess you you can have a pretty good guess about what's happening but you will not be able to exactly let's say define the exact quantity or that's knowing what's going on let's say you cannot draw that scientific conclusion in the end you have you only can describe what's happening yeah and let's have you explain also that when you are aiming to use tiling light sheet microscopy to image a piece of tissue okay that there are already a tremendous amount of points that you have to modulate the laser to image so let's talk about the number of points that you think are in like even just something as small as the fruit fly brain right and then next after you teach us about how many points there are teach us about how you then it's based on how many points there are and how much data is stored is based on yes the the nanometer of resolution right they're capturing that i also teach us about that process so actually your question also related to my later career decisions so let's explain this trajectory then because this is to west lake now yes yes so so i invented this technique at the stony brookie university right and it was patented and actually that pattern was granted this year so after i developed that technique it was pretty successful applied to some small specimens like some embryos etc but of course embryos are pretty simple simple multi-cell specimens you do want to understand a more complicated life right for example mouse brain ultrasound the brain how they think how they make decisions etc and that however there is a big another big problem you have to overcome is let's say scattering of the light or let's say both scattering and aberration of light let's say basically the light cannot penetrate through tissue right it can only penetrate more maybe several hundred microns and then it's completely scattered you won't be able to get any high resolution information from them based on the technique we have today and then i was actually pretty i was kind of disappointed at that point because i thought i developed something quite important however if you only can apply them to for example cialic ensembles or to the zebrafish at the best then your technique has a pretty limited impact right because you cannot use it to study more complicated animal models means right and the impact is something we have been looking for but as a point maybe in 2016 from a collaboration i learned about technique called tissue clearing technique so multiple labs are working on that technique so basically with that technique you are able to make tissue transparent although they are fixed so they are dead specimen but they are transparent right if they're transparent means you can you really can start to use light to get the 3d structure of those tissues which is something very difficult to do i mean all almost impossible to do before this technique was developed because before that what the best people can do is people have to cut the tissue slice by slice of course you can imagine amount of work you have to do and the mutations on them so basically not many people are doing that and then with this tissue clearing technique you suddenly have the possibility to apply the latest 3d high speed 3d high resolution immune technique to gain the structure of the all kinds of tissues right for example your brain may not mean not human brain yet but for the mouse brain organs and you can study tumors etc so many different possibilities and then i say wow this must be the future of this technique because it's limited the barrier of that prevents this technique to be applied to a larger field to make more impact right and i can see a lot of let's say commercial potentials of this technique and then that's why i decided to switch my direction career direction and and and also research direction at the same time so i after that i joined a technique company called 3i right and i uh in that company i i use my technique because i have everything in my mind about what instrument i want to build how i'm going to use my technique right and indeed i studied how to use my tiling light shin microscopy technique to to image tissues and indeed we mean basically it's a very natural transit by applying this tiling sheet light shin microscopy and then apply them on tissue imaging we can image large large clear tissue for example mouse brain because it's a centimeter size level right it's it's a very large size actually in in aspect to imaging with very high resolution however also the spatial resolution was great i realized several problems the first problem is um is uh the time you have to spend in order to get that information because the sample is so large i mean why you want high resolution right if you do a calculation you will find it's going to take hours even days in order to get a cellular level resolution of a few micron and because of we are using a tiling process tiling process is something as i said i mean you sacrifice the time to gain the resolution right but at some point that time that price become too high for example if you let's say for a small specimen if it's used to take one minute now you spend you spend five minutes it's okay because four minutes more right but if it's used to spend one hour now you spend it to five hours then that's a significant deal because four hours is a lot of time you don't want to spend right and then that's one issue i start to think about how to overcome and the next thing is of course about the the data size again because of the sample is so large and you want to get a high resolution typical size of data you have to collect and analyze this at least at hundreds of gigabytes level and very easily goes to terabytes or pb level level and then that's a let's say that these are the still frontier problems that unsolved and people are trying to solve and we are trying to solve and actually fortunately we this is something about our latest research actually about how you're overcoming these challenges okay if you imagine the process of tiling a lie sheet imagine this is a field view right i have a lie sheet i want to tell it one two three four times in order to get the entire plane image right that takes four times long longer time to image the plane hard also despite my resolution is higher but is there anything we how can we accelerate this process our latest technique called discontinuous lie sheet so basically instead of using one's lie sheet we are using a group of lie sheet for example let's say each of my finger represents a lie sheet right so if you want to if you only have one finger you want to put them in time positions but now you have five you only need to move them once and you can get everything down so that's our and by doing that you your speed can be improved dramatically right and you don't lose that resolution and you still keep that resolution and this is actually our latest result we were pretty excited about it and we get it submitted yesterday wow so but this idea was in my mind for a long time why i was why i was in 3i but because of the let's say the decisions in your company is always it's it's decided it's determined by profit right there's nothing wrong with it so in a company you have to make a profit to survive but how to make profit and how the decision about let's say there is short term profit then there is a long term profit and meanwhile i mean for any company you will need to decide what's the ending point of the company some company will be satisfied with let's say stay with 20 30 people and survive as as long as possible and then people will be happy with that kind of business but at some point i mean in my idea because think about at the beginning why i decided to do this tissue imaging was a blank area nobody was able to do it easily before now suddenly we have so many let's say good technique can do gain tissue structures um it's high resolution and we get we can get immediately get so much abundant information about our body right about all kinds of different tissues that information is a fortune i think it can lead us to a complete understanding in in in bioscience and in in relate to the our our health and for clinical applications disease diagnostic etc because it's tissue right and then that means you can currently you can connect this to billion dollar even trillion dollar business and that is the impact i think all the future i'm looking for i'm not sure whether it's going to happen or not but i think i mean there will be enough smart people see this kind of future and we'll be working on it and to me because i think i understand this technique and i understand the direction i would like to be involved in this process and i would like to develop the key technique with it and meanwhile i believe there will be other people working on different sets of problems to make the entire protocol executable on humans or on large areas and in the end if we combine our solutions together we can we can make something we can we can basically change things dramatically right we can we can we can make some pretty dramatic influence or change yes so then is it then taking multiple lasers at locations that are just far enough away from each other where you don't experience the issues with diffraction and yeah that's a that's a very good question actually that's a key question indeed you have to separate them far enough you can they cannot be too close to avoid in order to avoid the interference between between them and we do have a basically we use a a device called the spatial light modulator and we have some developed some good method to operate that device to generate the light sheet the discontinuous light sheet we want so it can behave in a in ways that we would like it to behave actually there are some more ideas we will apply to this actually which i won't tell you here but actually we can further improve the speed and resolution of this technique because even with this development if you think about to get high resolution result from mouse brain so our goal is to get to image in high mouse brain with less than 100 nanometer spatial resolution even with this improvement it's going to take years to have the entire sample image it's not fast enough we want to keep the resolution but we need the throughput to be higher and then you guys have ideas to make that which is what we are working on in order to make this thing happen why that is important because after you gain that ability you will be able to understand all let's say the goal is to understand how mind is generated in our brain right how decision are generated and you have to understand that neural circuit in order to gain this information it has to be fast enough to under because there are all kinds of different behaviors right so you have to make the throughput high enough to be able to image study all different types of behaviors and gain extract that information from the from the nature right from the mouse from other kind of model animals maybe fish right at some point maybe from ourselves so that's why this kind of ability is so important and we are going to put all of our resource in order to achieve this goal so that's something what we are working on and we think we have ideas about how to make it happen there are risks but the west lake is is one of the best place to that they gave us the resource they they they support us 100 to let us do what we want and make things happen that's why one reason actually I decided to join this university because again what we have discussed and you need the freedom you need the support you need the resource to to make it happen the part left to us is work hard yes yes west lake is a place of great magic of interdisciplinary magic yes it's beautiful and now you have two research assistants three phd's right now you guys are pushing the edge further and further in the field the frontier further and further this a discontinuous light sheet right right is a very interesting addition to tiling it's really interesting it's taking right now our imaging at as low as 100 nanometer resolution but when we were talking about this before we started in your lab right that this is still not doing the molecular side of things the the the the neurochemical right side of things and like something like the brain right so the imaging is is excellent and that's going to have to be overlaid eventually also with the neurochemical molecular side of things as well and that will give us the full picture exactly exactly so we are doing our job so I think this is a common goal of many research labs for many a lot of top research labs in in united states in japan genelia Harvard MIT I mean a lot of talk university a lot in europe a lot of groups are having this common goal and trying to achieve this goal right the indeed labeling is a very important issue we're trying to work on this each problems I believe many other people are working on this problem as well so we are willing to see what solution because I believe this there are so many people that are smarter than us they are going to give us new insights new solution at the same time we are doing our best and in the meanwhile we are collecting all the best methods for example a method called expansion microscopy we're also trying to use that so with all this everybody work together I mean as again come to our very first your your very first question right we all contribute the best part of of our efforts and by combine combining our solutions we may have this problem solved which is yeah we are excited to be a part of this this community and we are honored to be able to contribute yeah it is so cool being able to go to the different labs and basically learn how as you push a frontier someone else can take what you're doing and implement it in their life and you guys are just a couple hundred meters away from each other exactly and that's how you can create this this this community of frontier pushing faster right yeah right and that's such a beautiful yeah yeah yeah I believe because I have been following research works of many good labs I think there are people following me and yeah actually it's a small it seems to be a big word but in this case it's so small yeah so to me I mean I don't feel I'm at a different location of the world I mean my feeling is I'm still in this society uh that connect with each other so and let's also explain how you for the for the technology that was made with the tiling light sheet microscopy it's available for non-commercial use yes this is a very important so scientists from around the world can ask you for permission yes exactly design the software everything the whole process right right right yes I think this is our responsibility obligation to contribute to the society about what we know what we learn because let's say first from my own aspect we are developing tools and methods right I mean in order to make let's say 90% or 95% of methods dying in nowhere nobody knows the existence of them nobody remembers them they just disappear for scientists who are who is developing method you don't want that happen right you definitely want your technique to counter positively to scientific research right to make that impact so that's one reason I feel I want to push my technique to more research labs I have that belief that we are doing the best the immunity technique and I want to want people to take the benefit of our research so myself we can satisfaction from this process right the second is we are obligated to to do this because again mostly community is funded by private donors and some tax dollars as as well I mean this in my opinion this is a long-term investment of the society to scientific research because what's the fundamental reason that people are doing research it's curiosity it's there's not much say I let's say very few good researchers are funded or let's say produced because we know what's the what's the return right a lot of them just developed I mean or established based on our curiosity but we all know that our curiosity in a long-term in a long run going to return our society going to have a return that is unpredictable right that is kind of investment that is let's say the contract we are holding with the entire society so for that reason indeed our techniques be be distributed through two ways right commercially and non-commercial distribution commercially we do have new technique patented and licensed to companies to to make it happen because for some people even they have everything they won't build an instrument it's out of their focus it's out of their ability and they don't have time to spend on that they'd rather to to buy instead of spending their time to to build because time is money right time is for a lot of people more valuable than money but for other labs for many other labs they may use it for different purposes that's why we are giving our entire design software everything to research labs for non-commercial use we are happy very happy actually we are honored to to do that actually this is also something i learned from my post advisor arab basic in genelia he's doing that i mean i'm happy to inherit that tradition and to basically to to return of knowledge to the society not only to to china i mean to to everyone in the world i i think uh yes that that's that's our belief i love that part about your vision it's so important to communicate that around the world as we push the frontier of science we can open up our technology and tools that we've pushed the frontier with to enable thousands of other people from around the world to use those to keep pushing the frontier to advance it faster and to democratize the benefits across the world to all people right let's talk about the democratized benefits as well you told me that it would be so cool to see this technology be able to be used and approved for health care purposes around the world right how cool would it be to be able to see my tissues my biometrics or even our family members because most often it's our parents or our grandparents that have serious health issues and then we say scientists doctors please save my family and so the idea is that maybe when you have some sort of an issue going on with your body with your family's bodies you can use great technologies like yours at the hospitals once they are FDA approved to be able to take an image those parts of our of our health and then be able to do things like run artificial intelligence calculations to identify patterns predict pathologies from developing all different types of stuff tell us about this big picture vision for uh for augmenting our health so this is uh let's say first of all this is kind more less like a gut feeling right so why why I think this going to this kind of ability let's say collect um high resolution tissue information from human body going to mean something if significant because we don't know our body very well I mean with this kind of knowledge about cellular structure even cellular structure of different human tissues of our body we are going to learn a lot of things about our health condition about um genetic functions about why we are we behave differently etc we don't know right because we never had that information but from our experiences it's always like when you do have more information uh you have the chance to understand more about yourself about let's say about life so that's a belief in my mind so I'd rather to take that risk I mean of course I'm trying to sell in this concept at the same time and of course at this point we do have the chance to do it now because of tissue clearing because of a high resolution 3d imaging of course there are several problems you want to overcome at the same time right people will say well I mean why people would like you to cut their tissue to do this kind of analysis are hard to label them right I mean actually there are people working on these problems already I mean I cannot tell you exactly for example at least we know we can cut your skin right I mean people if you can skin cancer is is one of the most dangerous cancer among the world right especially in the US I mean if I don't think people would mind if you cut a small piece of skin and be able to know his chance to to to get skin cancer etc right I mean with ability we can do that for example when people are doing surgery I mean people won't mind if you cut a small tissue I mean it won't hurt them right and then there are a lot of ways to do that and I believe people going to develop better protocols and buy those doctors or biologists I believe there are enough smart people to have this have this problem solved and then what we can offer what we can offer is we can image that that piece of tissue in minutes with the resolution that actually we can do that right now and we are our technique is pretty unique actually in in the in the way of application it's it has a very strong self-alignment ability so basically that instrument makes sure it works by itself and then why that this is important because people who are going to operate the instrument going to be doctors going to be people who never be trained to do scientific research and we think this is very important for this technique to be to be used in hospitals in clinics as well right and then how are you going to use that I mean let's say after you gain this analysis ability how are you going to execute this analysis ability I mean we are trying to create a network like social medium I mean our big plan I mean it's something it's pie in the sky right now but there's some definitely something I'm trying to convince them people to do is I mean when we have that ability matured we are trying to push it let's say we are giving this technique we are giving this instrument to hospitals to clinics free I mean and then we are we give them standard protocols so everybody can use the same protocol to collect information and we are trying to build a cloud system based on all this instrument imagine if you have a thousand I mean there are maybe 20 30 thousand hospitals perhaps my number may not be accurate in China or maybe there are more hospital worldwide right if all this hospital has one a couple of this equipment and they work mean routinely high efficiently mean in minutes level to collect this kind of issue this kind of information from different tissue from different disease the tissue etc different organs etc we will have huge value very valuable database about ourselves about human and then I believe we are going to get something very important information out of this database mean that's our big goal I think I think this this is this going to be the fortune this going to be future of this technique the application of this technique at the human health level at disease diagnostic level etc people may have even better ideas at that point in the time of course when we are talking about cracked tissue let's say structure information from from human beings it's going to have a lot of other issues for example the privacy right and how this kind of application can be regulated well and how the information can be used wisely etc meaning on this is another thing that going to require better understanding communication and regulation between different labs different countries and scientists right so that's something mean it is highly important it's exciting and it requires collaboration requires vision requires passion and that's something we like to do yeah open up the big data silos around the world and help make the pattern patterns be able to be recognized better have the constant stream of biometrics from our health be able to be analyzed to predict pathologies I wanted to see if maybe there was a way like you're with your mass spectrometry taking it down from 100 kilograms to 5 kilograms if maybe there was a way to do it with tiling light sheet microscopy where instead of having to do it in the hospitals I could basically maybe wear something around me with me all the time that would dynamic capture in the dimension of time exactly first of all let's say we indeed we are working our instrument is not expensive let's say for some basic models the raw cost is about a hundred thousand dollars that's a it's not a small amount of money but compared to most scientific instrument it's a cheap device and itself has a lot of function secondly we indeed have solutions to make it even cheaper so we indeed can give it out for free right that's a very important thing but the last the most important thing is if it's indeed mean so valuable I mean nobody really care about the price I don't think price is the first concern when people decide to do something right for those big companies private companies when they decide to put billion dollars of investment on something for example Facebook Google or Apple when they decide to invest on something I don't think the profit is a big factor but profit is not perhaps not the most important factor I think impact is the is the most important factor because let's say money at some point is very important but after some point money is not the important issue at all for for for many for many people for many companies or our countries because money at some point it's it's paper right it's printed paper money is printed based on our confidence on our economy our behavior and that's our behavior is actually the basis of economy right if we need a certain amount of basic needs exactly that we want to start spreading impact exactly so I so I at that point I mean if we indeed can prove this is something extremely important available I don't think we will be lack of support or money to do it so in yeah after all we have to prove this is this is a good way to go this is something we can spend our time and efforts in yeah yeah how about on the way out let's ask some questions how can we inspire more people to collaborate together around our world yeah that's a that's a good question everybody should ask themselves I should ask myself me because sometimes people are mad at things for no reason right you're mad at other people mad at your family members you're mad at yourself at some point so let's say yeah let's go back to our first discuss first question again about we talked about the regulation right I mean basically perhaps we all have to think about what's the most important thing in our life what we are looking for what's important because the reason to make that question clear is because after you know what's exact thing you want what's important to you you can get rid of the disturbance easier so you won't be disturbed by many trivial things anymore and when you are not disturbed by those unimportant trivial things when you are very focused on your goal I think you have better control on your temper you have better understanding on people and you have better motivation to to work positively or constructively with people from different background or mind so I think that's a very important thing about how we can collaborate or work together better I guess in order to make that happen you have to understand yourself better right you have to understand what's important to you what you are what you want and what you can compromise in this kind of collaboration right that's perhaps the most critical part and of course in this person that's some other education concept we're trying to let's say embedded into our school is we have to learn that when we work constructively there there is winning situation right so basically one plus one could be larger than one we can if we indeed work together with good understanding we can we can make things happen uh tremendous the faster and nicer right grow the pie for everyone exactly which is a very very important thing the second important thing is if you have the belief of winning situation then negotiation how to negotiate how to resolve disagreement is another important skill we all have to learn right so basically you have you have to understand yourself better understand your partner better and to have the disagreement solved in a way that everybody may be have to compromise something but you also gain what you want is through that collaboration so basically understand yourself try to understand others and have the good belief let's say a common belief of win-win situation I think is the basis of of the interaction between different people and between different countries what widely I think yeah I think indeed in this in the world today I think this is this is very important concept that everybody should learn should yeah include ourselves I guess we should yeah yeah likewise yeah that's really great how about what do you think is the meaning of life this big human experiment right so yeah this is a this is a good question because at different period of time maybe we have a different answer to that question right but as this specific moment I think I was let's say when I was a student I trying to get into a good quality I think perhaps that's a good meaning of my life and then when I was in college I was trying to find a good job and when I was got my PhD or I trying to get a good payment a good salary perhaps that's very important to my life but at some point I mean none of those let's say none of those are the pursuit of those goals going to make going to last very long or going to make you're happy at some point I realized perhaps I mean my career is a little bit complicated I did a lot of different things I think in the end I have been looking for a better myself I have been trying to reach a better performance actually there are a couple movies that I like and I like their concept about let's say looking for a better performance looking for a better version of yourself because we have a limited life I want to see what's if I train myself if I did in a way that I think I regulate in a way that I think I'm going to achieve the best I want to see what's the best I can achieve at that point right so there was a good movie about this car the free solo I perhaps you watch it right that that guy trying to climb over cliffs I mean without any protection it seems to be crazy guy but I think I can understand him because he said some people look for happiness in their life some people work for look for performance right yeah I think I'm one of them looking for one of the guys looking for performance that's I think motivation can last longer and make me excited and another movie actually maybe extend the topic go too far it's called gladiator so gladiators if you at some point people will think gladiators are slaves right they fight to death it's pretty sad life but you but if you think about look look at the history if you think about them mean at some point they don't feel sad I mean they are pretty they are they are dying in a glory right so at some point if you think about the meaning of life everybody going to die at some point right you I mean you don't know what's going to happen to your life you don't know when it's going to be the end of your of your life right and then when those gladiator fight gladiator fights what's in their mind they are thinking about the death of glory right so what they are saying is he'll say the people about to die salute you right I think at some point we are all gladiators at the point so we salute to our lives right we want that glory in the end so that's the meaning of life yes oh it's beautifully said okay and then what do you think consciousness is you mean how that happens yeah yeah that's the question actually we are trying to understand right that's why we are doing all this kind of yeah this is very interesting because you think about what's the difference of uh yes so if you think about ourselves what are we we are a group of molecules right we are a molecule assembly that's all right that's all but at some point these molecules have mind has consciousness as you said we are trying to do crazy things and um and trying to understand ourselves right that's indeed a very interesting thing so basically again as we discussed a little bit as so it includes two parts right it includes a execution about the chemical reactions about how the signal is transferred by electron or by ions etc electrical signals and it includes another part of which is the algorithm that the logic that dominates this execution execution right and then this part of basically the logic and execution of the logic form our mind form and direct our behavior right and then where this is this come from and whether we can understand it in the end I think I have no answer I don't I don't know although we are trying to understand it um but indeed this can can drive I mean can drive it can drive our behavior for for a very long time I've been driving our research motivation for a long time but in the end if at some point we indeed understand how it's formed how it's created and uh might be I will be a little bit upset at the point I think because then what else you can do there's plenty more to push the frontier right after that yeah right what do you think that we are in a simulation yeah and you mean about metrics right I mean I don't know but what's the difference it doesn't make a difference right that's a that's a that's a is that a serious answer I've not actually I like that movie I indeed like that that aspect of perspective of rethinking our life but we cannot I think we perhaps cannot rule out the possibility right because our understanding is so limited I mean if yeah I mean we think we know a lot of things at this point but if you mean we are I think maybe human beings are always at this kind of exaggerated status we think we know a lot we actually we don't know much and we become arrogant at some point but perhaps we are right yeah if you indeed even by the even for the question you just answered right how much we know about ourselves not much right we really don't know I mean maybe it's just the start of a of a long journey and we don't know if how long this journey going to be able to be last right because meanwhile people are doing crazy things I mean people there are people doing positive things there are negative things happening eventually it's quite chaotic society on the other hand so yeah maybe that's why life is interesting right but um yeah it's not why people some people start to think positively people some people start to think negatively or destruct this constructively destructively I mean yeah all these are good questions worth of study I mean yeah in the end the research I guess that's why I think research is not about it's not about science it's not about technology it's really about the curiosity understanding our reality exactly it's I think that's a fundamental reason of of science is based on our curiosity of course I think for us it's a it's a luxury I mean it's a it's a yeah indeed I mean we it's our how to say that um so we had this opportunity the given opportunity to have a good environment to be able to answer these questions we have the resource to be spent to to do experiment we are asking and I think this is extremely on precious opportunities that we should really use these resources well to yeah to to give a good answer on them yes and then what do you think is the most beautiful thing in the world most beautiful thing yeah yeah so many what are they I mean perhaps life right perhaps life at some point if you ask me uh what's beautiful thing my what's reflecting my mind immediately uh are my children's uh yes so you see how they develop from nothing and to a life to a human that has a lot of uh thinking have their own will mean I I think this this this is this this process is so so beautiful yes yeah perhaps our life is a beautiful most beautiful thing of this world I think yeah indeed right that's we are what we are doing right I mean indeed there's no more more complicated thing than than ourselves and I mean exactly um it's it's magic yeah yeah yeah yeah yeah thinking about our our children as just this cell that is goes and becomes a living breathing thinking exactly being that has a meaning in life right right right right it becomes a part of yeah yeah you still remember the days they are baby and now at some point they can argue with you they can they can they can tell jokes with you and they can yeah I mean and they have different personality and actually yeah very yeah I mean that's that's awesome that's amazing for yeah my boys me my eldest son especially I mean he's such a interesting person I mean I just enjoy the conversation with him a lot of times I mean yeah so yeah indeed this is such a beautiful thing yes yes man thank you so much thank you so much for having a nice conversation huge pleasure thank you thank you yes and thank you for all your incredible work thanks everyone for tuning in we greatly appreciate it we'd love to hear your thoughts in the comments below on the episode let us know what you're thinking have more conversations with your friends families co-workers people online about tiling light sheet microscopy about these new optical methods that enable rapid high resolution 3d imaging of expanded biological specimens and also check out the links in the bio below to Westlake University check out the links in the bio below to Leungau's work as well check that out and also support the artist the entrepreneurs the organizations around the world that you believe in support them and help them grow you can find our links in the bio below as well you can help support us we can continue doing cool things like coming on-site to incredible places like Westlake University and conducting interviews with some of the most brilliant minds and go and build the future everyone manifest your dreams into the world we love you very much thank you for tuning in thank you and we will see you soon thank you that was so good thank you so much thank you thank you thank you