So there's been something that I've been struggling with as I think about growing this channel And I want to share that with you today because it is going to affect what types of videos that I'm putting out here the types of content that I'm creating and I just want to give you some direction into where I'm thinking about taking this channel and how I'm planning on growing it moving forward so I've been thinking about this for a while and It really became apparent to me in the last week that I need to make some changes here If I want to continue growing and if I want to accelerate that growth and grow faster So I'm gonna jump right into it. I was at dinner and I was with Jackie Who is my girlfriend of 12 years and she's my fiancee. We're supposed to get married last year, but COVID hit so we were talking at dinner and she had watched one of my recent videos and She made a comment.  And she said it seems you have a YouTube voice and I was like a YouTube voice. What do you mean? She's like it sounds Professional it sounds like a presentation almost like a commercial and she wasn't she wasn't saying it as an insult She was just making an observation So she was just watching a couple of my recent videos which are edited Which do you have more of a commercial feel and she said it doesn't sound like it doesn't sound like you There's no conversational element in there. It's just it's more of a presentation And again, she wasn't saying it in a bad way, but she was just noticing the recent videos So that really struck a chord with me because it's always tough for me to get into filming mode I am naturally an introvert. I like to be alone I like to gather my thoughts and when I have to get into high-energy Film videos on YouTube mode. It's tough for me. So I've always wanted to make videos that weren't so stiff I wanted to make it more more real and authentic and personal and More than just tutorials and guides on online business, which I'm passionate about online business I love online business and entrepreneurship But I've been wanting to make videos outside of just tutorials and how-tos for a while And I've been thinking about how to change that up So I have a few ideas now that I want to share with you today But first I want to go into to my story if you don't already know me and if you are coming here for the first time This will probably be the main video on my channel if you're new if you're a new viewer My story is I worked nine to five as a project manager for three years after I graduated college I'm 30 years old down I graduated in 2013 and I graduated with 60 K about about 60 K in student loan debt But was able to pay that down in two and a half years just by being aggressive with my payments And also I was able to build an online e-commerce business that gave me a second income So I was able to pay down $60,000 in student loan debt in about two and a half years in 2016 I was able to quit my corporate job After my econ business started making a lot more than my day job and from there I built up that econ business to multiple seven figures a year I was able to sell it in 2018 and from then I created two other online businesses one was called growth response and That was a services company where we ran legion services and cold email services for other businesses And I also have an online education company called productized profits Where I teach people the skills that I've learned that allowed me to build my online businesses and make a full-time income online So there's nothing I enjoy more than just sharing what I know and teaching people what I learned From being able to build multiple businesses online to the to the multiple six to seven figure level And I love helping people develop these skills that allowed me to quit my job make a full-time income online Have the freedom to travel whenever I want to wherever I want and be able to invest and do things that interest me And I'm gonna continue doing that. I'm still gonna teach. I'm still gonna share what I know But one thing that I've realized about YouTube It's not just about education a lot of it is to do with entertainment And a lot of it is to do with having some kind of personal touch and giving people a reason to come back to your channel Right something authentic something raw something that shows more about you as a person because people come for you Right, they stay for you and they come back for more and more because it's you so that's what I've been thinking about and What does that mean for you? Well a few things future content here is is gonna be a little different I'm still gonna do tutorials and guides and and teaching you skills about online business and creating internet income That's still the core theme of this channel. I don't plan on changing that But what I do plan on changing is adding more styles of content around other things that interest me. So It could be things around personal finance Investing it could be trending topics that I'm curious about things like crypto, right? I wasn't I tried to get back into crypto earlier this year. I initially got in 2017 and it stressed me out because it wasn't just business related But to me that's that's really interesting to me especially right now as we're seeing crypto start to rise and gain popularity again So I want to be able to talk about these topics and other things that interest me And if it fits into the theme of internet income, I'm gonna do my best to tie it in if it doesn't I'm still gonna do my best to tie it in that's still gonna be the core theme of this channel is creating internet income and Developing the skills needed to make a full-time income online But I want to be able to talk about these other things that interest me That would be beneficial to you as well personal finance investing things like crypto trending topics Maybe just my personal opinions on things that are happening in the world So that's what I plan on doing with the content moving forward now My favorite follows here are people who keep it real anyway people like Alex Becker like tech lead like Olier Like there's a guy named Joshua flute that I recently found. I love their content because it's authentic. It's real It's not commercialized. So I'm gonna start creating more content like that And Hopefully you stick around if you don't I understand because I've created just mostly how-to style content so far But I plan on still doing that, but there's gonna be a mix of other things in here So I wanted to share that with you. That's the direction. I'm planning on taking this channel moving forward I think it's gonna help me grow and also help you learn a lot more from me in my Experiences and be more versatile and learn more about things that that you might not be thinking of right things outside of just online business and Just tutorials around marketing. So just want to drop this video to let you know where I plan on taking this channel moving forward I'm excited for this. I hope you're excited as well I think there's gonna be a lot of cool things coming down in terms of content I have a lot of ideas moving forward now that I can kind of branch out a bit So I hope you stick around I understand if you don't but I think it's gonna be a lot of fun So that's it for this video and in the next one. We're gonna get this kicked off I have a how to get started with crypto guide coming soon where I I'm starting to invest 10k and seeing how that goes So that's coming next week. I'm looking forward to sharing that with you and talk to you soon. Take care


Hey guys, how you doing? This is Rich from RichTV Live. I thought I would just go live since I'm in like this beautiful place and there's like all these 100% by top 1% signal strings that were popping off today so I thought we would just talk about it a little bit. This is beautiful British Columbia. This is Vancouver. These are the views. And yeah man, when you have 100% by top 1% signal strings popping off all over the place and then you see news that BMO puts out a buy rating on Supreme cannabis, Supreme Pharmaceuticals, Supreme Cannabis. And they give it a $2.50 buy rating and then you see Village Farms and it just doesn't want to stop. It just keeps going up every single day and that's 100% by top 1% signal strength. And then we got Aurora Cannabis which I've been talking to you guys about every day and they continue to go up every single day and that's 100% by top 1% signal strength. And then there's CXXI another one that our community just keeps talking to me about and they keep exploding every single day and they're up 100% by top 1% signal strength pretty much or like 96% by something crazy. And then you get T-God and then T-God goes up 100% by top 1% signal strength and T-God explodes today over 5 bucks. So you got all these 100% by top 1% signal strengths. Now the question is which one are you gonna buy? And then you got MedMan that is climbing up secretly coming from a bottom and they've been steadily climbing up and they were a 72% sell then they went to a 40% sell and I believe today they're now a 32% sell so they're starting to build momentum on the opposite side of the buys. Your loving thank you you like the view your loving your T-God and Aurora you should be they're two absolute monsters in the cannabis space two of the biggest companies in the world in the cannabis space so you're doing pretty good with Aurora and T-God. Both 100% buys top 1% signal strength HVT yes HVT was on fire today guys harvest one I've been talking to you guys about harvest one for a couple years now they're absolutely on fire absolutely on fire guys HVT it's on a full alert full alert for HVT guys full alert still holding acreage yeah acreage is a beast too you should be holding acreage absolute monster I gave you guys some news today about fire Supreme cannabis I'm telling you guys SPRWF fire in Canada it's going to explode it's so cheap right now and the fact that they work with Wiz Khalifa it's crazy there's the yachts are down there yachts are right there Zenibus is a great buy Zenibus at 333 is a great buy you're gonna do very well at 333 with Zenibus who else was on fire today that I'm missing you got 10,000 shares of fire last week yo genius you're genius no it's a beautiful day here in Vancouver it's 18 degrees right now spring is here spring has arrived here in BC you guys see that I think fire is going to explode I told you guys T-God was my number one pick at 2019 and I'm not really surprised that it's exploded Seneva is gonna be another monster because they got a tight float yo Jared what's up man you know where I'm at you know where I'm at a boat ready to get on a plane and go do some business sold VFF today huge move from Friday's low yeah man great move great move Village Farms has been an absolute monster hey guys we got 47 people watching 10 likes come on guys you can do more likes than that you can get more likes than that one of the biggest youtubers here in Vancouver at least you can do is hit a couple more likes I'm at the top of the mountain here man who else is giving you guys these views talking stocks and giving you guys these views who else does this I don't think anybody else can do this I don't think anybody else can pull it off 21 likes wow 24 likes yo you guys are the bomb man love you guys guys absolute bomb man so just a lovely day here in Vancouver British Columbia this is why we have the most expensive real estate in the world you want to know where the expense of real estate is it's on those mountains see those mountains that's where the expense of real estate is those mountains up there West Vancouver North Vancouver that's where the expense of real estate is if you live up there with the view of these of this ocean if you're on the mountain with the view of this ocean you're killing it you're just killing it yo thank you guys for the 33 likes a mad respect to you guys man see if we can hit a hundred likes see how fast we can hit a hundred likes lovely day with ACB teagot and Hexo yes Hexo I forgot about Hexo Hexo is on full alert guys they had a monster day today man I bought Hexo last week sold it too soon and they exploded today it's always nice to know you're on the right track just sucks when you sell too soon what am I watching for tomorrow tomorrow I'm gonna be watching Hexo I'll be watching Aphria Aphria had a monster day today I'll be watching Aurora I'm gonna be watching Eve Eve and Co had a monster day today up 17% I bought myself 10,000 shares I'll be watching Village Farms I'll be watching Medman Blisco had a little bit of a sneak attack day today they seem to kind of climb up when everyone least expected it no Vivo's not dead man love Vivo it'll have its turn every dog will have its day every dog will have its day but we're in the right track we're in the right lane I literally sold too soon on all my picks last week I could have held Village Farms I could have held Hexo and I could have held Aurora and I would have done better so at least it means that I was in the right lane I just got out too soon I got out I got out an early exit I got out one or two exits too soon that's okay I'd rather get in too soon than too late you don't want to miss your exit and have to turn all the way around right I don't want to do that so I'd rather get out with a profit today I was up on Medman like 200 bucks and I was up on Eve like early on like a hundred bucks I could have taken out like 250 300 bucks between the two I made 250 today on my Aurora trade and I could have sold that to try to you know hit my goal of at least 400 a day today but I didn't want to rush it I was like you know what I'm gonna hold those because they got massive momentum and let's just see if that rides until tomorrow I don't want to get out too soon so let's see Nick you took 24% on Aurora good work man that's huge yeah if you guys want to see a plane check this out check this out come with me I'm gonna show you guys around don't be shy everybody's friendly here you know you're up 55% on Aurora that's big time man big time yeah you're 10 minutes away bro what am I thinking for T got tomorrow with the release report I'm expecting big numbers man expecting big numbers yo it's getting loud here too so taking off just like your portfolio it's taking off and just like all the cannabis stocks taking off AI PR like I PR man it's a beast absolute beast I like I PR but yeah I'm not I don't need to buy a stock at 78 bucks 80 bucks whatever man we can find cheaper stocks than that that are good companies hey guys we got 76 people watching 46 likes let's see if we can smash a hundred likes guys we've been live for 11 minutes we're in downtown coal harbor Vancouver first Columbia Canada one of the most beautiful places on earth right here ranked as one of the most beautiful places on earth and best places in the world to live and that's where we're at right now check it out check it out always ranked one of the best places in the world to live you guys want to know why we got the mountains and we got the ocean that's why you got houses on the mountains those are expensive I believe T God will hit $20 as well which hemp stocks do I think we're moving up I mean I see so many stocks moving up hemp CBD like everything guys like everything that we talk about is moving up I mean if you want to know which stocks are moving up just watch which TV live you'll know which stocks are moving up it's pretty obvious no man I had village last week I bought a thousand shares and then I sold and it hurts hurts hurts but it is what it is am I gonna see playing it all the way I don't know stay tuned man stay tuned you have no idea where I'm gonna be where I'm gonna go who I'm gonna be going with that's the beauty of rich TV live it's always a surprise there's a lot of people watching right now like who is this guy people around me like yo who is this guy who is this guy how high do I see fire going I think fire will be a $10 stock in the future I can say that pretty comfortably if they don't get bought out first full alert baby it's always a full alert it's always a full alert when it reaches in the house you know it's a full alert my thoughts on Hexo I bought it last week I think it's an absolute beast had a crazy run today and I missed out on it and it hurts so yeah I like Hexo man they got Molson's and now they got hit who supposedly I am this has been sniffing around TGIF you know what that seems like a perfect partnership yeah G A G J A G X was on fire today 193% up doing a dead cat bounce you bought some meal today I hope you win on that man so yeah there's just like people everywhere enjoying this beautiful spring weather people from all over the world here in Vancouver and we're live with 78 people emerald health is under the radar has a long shot I think it has a lot of potential MC IG yeah I know MC IG my e-cigarette see you later George so yeah this is Vancouver guys I want to talk to you guys a little bit about Supreme Pharmaceuticals Supreme Cannabis Company I think it's going to be a monster I brought you guys the news brought you guys that alert so take a look at it everything else that I've been bringing you guys 100% buy a top 1% signal strength we've got lots got lots that we're looking at look these guys these guys are jumping around see if you guys want to be on YouTube yo what's up guys what's up we got 74 people watching right now Vancouver is kind of quiet if I was in the Dominican it'd be like party right now wherever I am at this time and Coopers like beautiful sunny day and everyone is so polite everyone's so quiet even the guys that are jumping everyone's peaceful here even the guys that are jumping off of their scooters are quiet about it look oh nice that was a good landing you guys want to see this check us out can I make some videos for you guys for YouTube yeah I'm live streaming right now I got like 72 people watching so if you guys got any cool tricks I got 20,000 subscribers it's rich TV live so we're like literally live right now I got people watching right now 74 I want to see some tricks you guys wanna see some tricks see some tricks you guys gotta show me some tricks man don't hurt yourself though oh nice nice nice nice I was pretty good what would you guys rate that out of 10 I was pretty good bro that's pretty good that's pretty good so what's up man you know this is YouTube YouTube oh what yeah we're live on YouTube there's 78 people watching right now my channel is called rich TV live look that up later we got 20,000 subscribers holy moly yeah man I'm talking about wheat stocks that's why I got the hat yeah what's up so what's up to YouTube so you do yeah you got any cool tricks you can show us hi yeah don't hurt yourself for anything but if you guys got any cool tricks we got some people watching we got people watching okay we're watching we're watching here we go oh the ranking you guys we got an eight we got a ten oh whoa whoa yo that was crazy yo did you guys get that yo that was good bro yo on Instagram Bryce underscore gravy B R I C E underscore D a V you follow back to yo that was a good trick man I like that trick I wasn't expecting yo they got a 9.5 on that they're giving you a 9.5 for that 9.2 9.5 9 my YouTube is rich TV live yeah there you go drop your IG bro drop your IG what about you you got any IG you want to drop drop it yeah Jaden underscore have it there you go man right here that's me bro subscribe yeah thanks bro not respect man I talk about wheat stocks if you want to make money just got to go to rich TV live man subscribe like you did hit that bell for notifications all right I'm always around town okay good work guys if you watch the channel you'll see your videos up there okay all right please just like that man just like that guys it's just like that that's how it is it's how it is man so all right we're gonna make a move guys I'll keep you guys updated on what's happening yo you want my like jump what jump what jump what you want me to jump off one of those things yeah yeah Jared already knows what will happen if I jump off one of those things I'll break something as I should ask me to jump all right guys I'm out peace fire supreme cannabis company fire fire fire supreme supreme supreme almost peace


Today, I'm very excited to host a very special guest, Dr. Kirsten Switzer. She is an archaeologist with a research focus in ancient magic and ritual practice. She's doing fantastic forward-facing scholarship, bringing this work to a broader audience online. She's currently working on a book about magic, ancient magical signs. So Kirsten, welcome to the show. Thank you for having me as your guest. I'm totally excited to be on your YouTube channel. I love it. Oh, the pleasure is all mine. I'm a big admirer of your work, love your channel, learn something new every video that you put out. And I just want to thank you for your scholarship. It's really a great service to the public. It's so kind of you. Thank you. It's OK. So, Kirsten, for those of my audience who are unfamiliar with the Greek magical papyri or the PGM, could you give a brief explanation about what they are? Basically, the Greek magical papyri or what's behind the term, the Greek magical papyri, is a modern compilation of an awesome body of ancient magical papyri from Egypt. They date between approximately the 1st century BC and the 7th century AD. You can differentiate them into two different kinds of primary sources, ritual manuals.  And the other primary sources is papyri, is artifacts like from applied magic. We have about 70 ritual manuals and I think published currently are a bit over 100 papyri artifacts. So these are the two main sources for our study in ancient magic and for research in ancient magic or basically for our understanding of ancient magic. It's the ritual manuals and the artifacts. And the Greek magical papyri, as the term says papyri, comprises all the papyri artifacts and manuals. But there are also much more artifacts from different materials like most people have heard or many people have heard of the curse tablets, which are made of lead and of magical gems. So these are really huge groups of artifacts. But the corpus of the body of the Greek magical papyri consists of roughly about, it's already about 200 Richard and artifacts comprised. So the Greek magical papyri were first published in the 1920s from Karl Preisenland, so the German scholar, and the first English translation was edited by Betz in 1986. And the Betz translation is what most people actually work with. Currently, the first volume of a new edition has been released this year by Chris Farrow-Own and Sofia Torales-Tovar. This is like they don't only publish the papyri that were known until 1986. So Betz in the Betz 1986 publication, it's 130 papyri altogether. And the new volumes will have like everything that has been discovered from then until now, there will be translations available in the new volumes, too. It's a modern compilation of all the papyri from Egypt we have about ancient magic. Oh, the Greek magical papyri we have from Egypt. The Greek magical papyri seem to be, they don't seem to have been very appreciated in history until the work of scholars like Albrecht Dietrich, you know, working on these and, you know, appreciating their values. So I didn't know if you could talk about just the reception up until modern times. The earliest modern or the early modern scholars working with this magical text. There are a lot of Germans who did this. And that was in the late 19th century and the early 20th century. And at that time, there was the perception was that magic is everything that's bad. It's bad, magic is black, it's dark, you know, it's all these curses and harming people. And there was this clear differentiation between religion and magic. Religion is everything that's great and good, and magic is everything that's bad. And actually, the first German scholars translating the Greek magical papyri, that was in in in official manners, like when Preisen-Dannes, who compiled the Greek magical papyri and translated them for the first time in two volumes and made them available to more researchers. When he taught this magic papyri at university, he wasn't allowed to use the word magic in the title of his lectures. He had to describe it, but he wasn't allowed to use this term magic. So the perception was for a very long time that this magical text that's not the you know, you have the golden classical grade text from ancient authors or the Egyptian pyramid text and religious types. And they were highly appreciated. And when it came to the magical text, it was like, oh, no, this is not even literature. This didn't have any value. You know, if you if you wanted to be someone in classics or in your research area, you definitely didn't study magic. So this was a very brave approach of the first scholars actually dealing with magic and translating them to translating the the texts. And it's really interesting to see how with the publication of the texts and making them available to more researchers, the our understanding and our approach towards towards magic changed. We came from this it's it's dark and it towards a well, you know, see, if you take a look at it, there's a lot of Egyptian religion in there. There's a lot of Judaism in there. There's a lot of Christian elements in there. You have this syncretistic approach and you do have very elaborate invocations hymns in there. You have texts that clearly they're very much older than actually the the the texts we have. So it shows that there is the knowledge, the people who wrote these ritual manuals and who created the magical artifacts were very well educated. You know, they were very knowledgeable in ancient religion and not just in one religion, but in in multiple religious beliefs. They were very familiar with with traditions, with so so much knowledge, traditional knowledge, this awareness. It came up just because Preisenthans and other scholars started to publish these magic texts, despite the opposition within scholarship towards the the magical texts. We went to like in the in the 80s, 1780s, we went to this discussion, discussion, what is ancient magic? And it was like scholars tended more to while there is not a hard line between religion and magic, we can't really differentiate this. And the tendency was there that, you know, magic is more like religion because you have very similar gods, you have similar traditions, identical techniques like like the invocation, like sacrifices, offerings. And a lot of the the the content, actually, what you said were were recitations from from older texts. So there was this idea, wow, it's all religion magic, isn't that separated? And then there was there was some kind of rise in the 90s in the study of ancient magic. And then that client now since around 2010, the the interest in ancient magic increased drastically and including discussions, new discussions about what is ancient magic and how, you know, we all use the term in scholarship, a lot of publications like ancient magic, magical artifacts, me included. But how do we define this term? Because as a scholar, we should be able to define the technical terms we use. And this is really, really not easy, not only from from the modern approach because that's what what has been a lot a long time. The modern approach to define ancient magic and what the the the the turn we we see in scholarship is like trying to understand magic. We have to do that much more from the practitioner's perspective, you know, from the ancient perspective. And as it is today, there wasn't just one kind of magic and there wasn't one definition of magic or one understanding of magic or one use of magic, despite lots of different terms in the Greek magic, a papyri for magic, which are all translated as magic. But behind these these when you when you read bets and you find the term magic, it's very likely that the original term is not magaea in ancient Greek term for magic, but there are a lot of different terms behind it. So it's a very ancient magic was very diverse, very complex, lots of different thoughts, perceptions underlying the application of magic. And on top of that, you had a lot of people practicing magic and likely all with individual approaches. It's all a matter of edic versus emic perspectives, right? Like, what do we even mean when we say something's a religion versus something's magic? We're defining borders, right? We're saying religion is what we do, but magic is what they did. Very much like an outsider or insider defining term, both religion and magic are anachronistic terms to begin with. You see a lot of bias. You know, the early scholars working with with anti-magical texts faced a lot of bias towards towards magic. And that you still have, if you read academic papers from colleagues, from certain colleagues closely, you still see a lot of bias. And even when they try to to avoid sounding like they despise is a bit tough. But, you know, they disagree with magic or they think magic is something negative or something minor. You can still see it in in the words they use to describe magic, how they describe magicians, how they address them. And you still have this bias, but it has become much better. There's many, many young scholars that approach magic with less bias, but with more curiosity, focusing on the question and not focusing on your values or your personal belief or your personal attitude, but really focusing on on research. Just kind of making a little cross-reference here. When we talk about things like Neo-Platonism and the classics, like you look at a philosopher like Plato and there are certain scholars who want to look at Plato or the Platonic tradition and define it in strictly philosophical terms and forget that there was a lot of mysticism and a lot of magic involved in that, as well, unwritten doctrines, things like that. John Dylan, who wrote the seminal book, The Middle Platonist, coined things like the Chaldean Oracles, the Greek Magical Papyri, Sethian Gnosticism as the underworld of Platonism, the Platonic underworld. So it just goes to show that bias still exists in scholarship as well. Scholars, classic classicists and pure philosophical-minded scholars would go. It's low magic, low. That's what the that's what the others were doing versus the. But what the reality is, is that. I think when you look at something like the Greek Magical Papyri, this is more of a window into what the everyday person was probably doing versus, you know, somebody like Cicero or somebody like Lucian or Clement of Alexandria, these elites, in other words, who are literate and had access to these materials to even write stuff down to send it to posterity. The people on the ground are going to the local ritual specialist, right, and getting their their their love spells or their spells to bind and unbind. You did that wonderful presentation on Patreon about those kind of spells, which I loved getting to the next point. And this kind of ties into what you were talking about. I just wanted to point out this quote that Hans Dieter Betts did make in his book, The Greek Magical Papyri. The religious beliefs and practices of most people were identical with some form of magic and the neat distinctions we make today between approved and disapproved forms of religion, calling the former religion and church and the later magic and cult did not exist in antiquity, except among a few intellectuals. So I think it's a good. Yeah, it's very it's almost like a mic drop moment there. What is the utilization of this technology? What are these rituals tell us about the everyday person? OK, so now that you drop that quote, I do have to address it. The first thing is when you hear most people, when you hear anyone, academic and non-academic, he is saying most people. That's basically an attempt to create the impression of authority.  And in terms of magic or the study on research and ancient magic, we don't know about most people and we don't know even most people about most people in antiquity, especially in terms of ancient magic. So what Betts is trying to do here is to I think he's he is to address the the turn of approaching of how scholars approach magic. But still, you know, if you if you study ancient magic and if you really want to go into the meta and understand it, you have to be critical of what you read. The other point is the eighties and nineties where what Betts or the definition of magic here that Betts says, well, you know, there is not this hard water and religion basically is for most people is some kind of magic. That approach is typical for the eighties and nineties. It's an entirely doomed to fail approach to have the discussion into the direction of saying it's all the same. So religion and magic, you know, is more or less the same. When the goal actually would be to understand the magic of papyri and ancient beliefs, thought and actions better, because in the moment you say, well, it's more or less the same or magic is is much like religion. You put it all in one pot. The reality is if you look at the Greek magical papyri, they are so diverse and so complex in thought, in belief, in however you want to define religion. But in terms of the classical gods, Egyptian, Greek, Roman, Hebrew, it's they are all there. So you can't just say it's it's it's mad magic is like religion.  That's that's very similar to religious practices. But if you look into the details so that the technical aspects are very similar. But if you look into the details, like how belief is expressed and what's actually the underlying thoughts of individual rituals and how they work, I don't know if you have ever had a discussion about magical theory, a theory of magic, but it's so diverse. There's so much different kind of ideas or magic works, you know, a discussion you don't have in classical religion. You don't discuss how a power, how God can do what he or God is what she can do. But you do have that in the in the magic of papyri. You do have explanations or you can see them when you read them, the text, the ideas and the approaches to explain how things work, how power works, how magic works. And that that's different to what Beth says. So I had to address this because we are we are slowly heading into this direction to say, OK, religion and magic is not in terms of religion is good, magic is bad, but it's also not in terms of it's more or less the same. And it's I think it's really it's much more important. It's really important to to open this door in scholarship much more to talk much more about this, that we're now at a point where we scholars accept that magic is not necessarily low level. And we are at a point where we see, OK, no, no, we should kind of level that in terms of our attitude towards it with religion. But now we can safely say religion and magic is not the same without saying magic is bad and religion is good. Just if we get back to it, like I think the only thing I'd add there is you're absolutely right when we put things into boxes that are needs like that, like religion and magic. We we tend to look at it from a standpoint where we don't recognize the diversity within those boxes. We tend to put a lot of diverse voices into this false sense of everything is the same when, like you said, there's conversations going on even within theories of magic and the Greek magical papyri about how things work, the theories in practice. It's very important to recognize that. Thinking about magic and what magic is, and especially if you look at the original terminology in the magic of papyri, it's it's like there is so many new doors that open up compared to the to the translation. The imagination and the the the underlying concept, the concept of the ancient people who practice magic or who wrote the magic of papyri is so so rich and complex. It shows so many various explanations. Just look at divination practice and how many different ways you you you can achieve a divination.  Certainly they were in frauds. I mean, there were as everywhere in every business. There were frauds, too. But the majority of the people and people who wrote the magic of papyri, they were really intellectually high level. You know, it's the highest grade of knowledge and very educated.  They they were people living in a very dynamic, cosmopolitan milieu. You had to be familiar with many different traditions, right? You had to be familiar with your own native Demotic traditions. You had to be familiar with the Greco-Roman traditions, that the Hebrew traditions, the Christian traditions, etc. These were very well learned people, especially if you if you keep in mind that it was still not easy to get access to books or to scrolls. It's not that everyone had the had the the financial means to own books or to own papyrus papyrus scrolls. Papyrus wasn't an expensive material and not everybody learned to write. You had to have access to not only the literature, but also to teachers, to, you know, to schools. And it's not something everyone had the chance to do. The other thing is when we look at the time, the majority of the Magical Papyrus dated around the third and the fourth century, 80. This is a time when more and more pagan temples, pagan, pagan temples get closed. By the Romans. So the question is what happens to the priests? And we know that, you know, in larger temples and temple complex, there could could have been 50, 100 or even more priests working there, living there. So what happened to them? How did they make their money when the temples were closed? I think that that the people who wrote the Magical Papyrus and who practiced magic, basically for what you said earlier, everyday people, you know, not only kings and royalty and for everyone who could afford more or less, these were very likely, at least in parts, partially priests. So extremely educated people, the intellectual elite in Egypt. This is something we should keep in mind, you know, who were the authors of these texts? Who could have had the knowledge that is preserved in the Greek Magical Papyrus? But not knowledge that is restricted to leadership elide, but or rulership elide. But that becomes accessible to a broad audience in terms of more people can participate in the benefits of this knowledge. I love what you just mentioned there about the disenfranchised Egyptian priests. This this is a very interesting parallel with Hermeticism. A scholar named Christian Bull recently wrote a book about who wrote these texts that we find on the Corpus Hermetican. He came much to the same conclusion that you did. These were disenfranchised Egyptian priests who were creating these texts. So a very interesting parallel there. Yeah, I mean, and it would all check out, wouldn't it? These these are ritual specialists. They're familiar with many different philosophical and, quote, unquote religious concepts. It would just make sense that they're playing their trade just in different ways, like in one sense with the Hermetica and in another sense with these magical inscriptions and rituals. And you also mentioned like the importance of the Pidea. Pidea was not Pidea in education. It was not an inexpensive venture for any family in this period of time and even access to a library involved, even in somebody's house involved, much access to access to much means you would have to put into that. I think the next part I wanted to talk about, just to have a little fun, fun here. I wanted to talk about some of your favorite rituals in the Greek Magical Papyri. I have a couple of favorites of my own. For instance, I really like the Invisibility Spell one twenty one two twenty two through two thirty one. And I like the Personal Diamond Spell. And of course, the very first spell and bets text, you know, where you create your personal diamond assistant and it just does everything for you. So I didn't know if you could mention maybe a couple of your favorites. That's difficult. So with the Invisibility Spells and the Personal Diamond, who's basically a Piedros, and this is the Piedros is one of the most common rituals in the in the in the Greek Magical Papyri. And a lot of people like or love exactly these two kind of or kind of spells. I think my favorite it's not that I could say it's this spell or that spell. It spells that provide in in in mostly in in in a site sentence, some personal information, you know, or some more personal information about the people writing down the instruction or making an artifact or what I totally love is when you see that someone writes down a ritual and then crosses a whole line out. And then the next line is almost identical.  You know, it's just one one letter is different. And you can literally imagine you can see the person sitting there writing and then swearing because something was misspelled. Or when when when he called, you know, you have an example in PGM seven where you get the impression that the scribe actually copied from from from from a sheet he had and then misread the line. You know, you slipped in the line, you know, you started the wrong line and then copied the wrong line and had to cross it out and then had just had to copy the right line.  And I also love the I love cryptography and and steganography and everything that has to do with encoding. So I especially love the ones like the Eighth Book of Moses. Everything at the beginning of the Eighth Book of Moses is, I think, steganography, you know, where where information is hidden. Normally, steganography is is not encrypted writing, but hidden writing. So we know it mostly of from from hacking and from from our technologies, where you hide information in an image. But the Eighth Book of Moses at the beginning, the secret, the whole Eighth Book of Moses ritual is basically about gaining knowledge of the secret name of of a very powerful God. And when you know the secret name, you can perform a lot of short rituals, which are very efficient. The description of the ritual starts with images, with literal images, not with painted images, but images painted by words. They are, I think, steganographic. There is the real name is hidden in there. And the the way the there is an artifact that has to be inscribed. And the the the description of what has to be inscribed is basically encrypted with words, but not in terms of a code, but a picture painted by words. And this is what I utterly love. And we have one ritual manual. It's the first century currently it's dated to. It's the one with the first mentioning of characters of magic science. That has been entirely written in code. It's a monoalphabetic substitution code. So each letter of the alphabet is written with a different letter. So it's it's not really a high-tech cryptography, but it's absolutely awesome. If you think that a person wrote an entire ritual manual encrypted. This is what I love. And I think it's pgm 12 where we have the encryption of ingredients. You know where it said that when it's the eye of a snake or was basically it's mostly it's it's animal parts, parts of animals. And then it says what it really means is and then you get plants, plant parts. I talked about that before. I did the fascinating thing about this is that we have a lot of ritual instructions up until the middle ages and today which claim to you derive from ancient traditions and they use animal parts and blood especially blood, blood of snakes, blood of whatever. The encrypted Greek magic papyrus says that these were actually codes for plant ingredients, plant-based ingredients. So that I wonder how much of these animal sacrifices and blood and things that are used, you know, that are described in ritual instructions were actually code for plant ingredients. We don't have all the plain text, all the explanations for the codes. It's really interesting because, you know, at this point in time in history in antiquity, mysteries and secrets have a kind of social capital, right? It gives the spells, it gives the traditions a kind of prestige, right? Basically, the secrecy is fundamental existential fear. I could say an expression of fundamental existential fear because the ancient belief towards magic or in terms of magic is that it's not something you are gifted, you have to be gifted to perform. But the ancient idea is that magic is entirely based on knowledge so or can be learned from everyone. Keeping your knowledge secret was fundamental to ensure that you keep your clients. If everyone gets access to your knowledge, everybody can use your knowledge and, you know, can offer your knowledge. So you have this all over the magic, the magic papyrus where the author says, well, keep that secret, make sure you keep that secret, don't share that, only share that with your first son or something like that. So it was just a very pragmatic approach towards knowledge. I also wanted to get back really quickly to what you were saying, what you enjoyed about the human aspect of looking at these spells. I was reminded of one of the texts in the Nag Hammadi scriptures, the Prayer of Thanksgiving. It's this really lofty prayer and then at the end, you have the copyists writing out, I have copied this one of his discourses, a great many have come into my hands, but I have not copied them because I thought they were already in your possession. I even hesitate to copy these things for you, since perhaps you may already have received them and the matter may annoy you for that person's discourses that have come into my hands are many. It's like you have this really lofty ritual invocation, right? Then you have like this dude pretty much just like writing to the other guy, okay, I wrote it out, but I'm not sure if you have this, maybe you have it, maybe you don't. It just gives it a very human dimension that I love. Exactly, because you have the best points. You have this real person, we actually do have real letter preserved that tell us, the magical pirate tell us that this is a letter from, or I got this, I sent this to you, I had this sent to you, so that we learn that the knowledge was exchanged, among other things, through letters or by letters. People actually put, when we have one letter with a texture artifact, where the texture artifacts is described, so we know this is the way the knowledge was exchanged and that is not really very mysterious. And it's not a secret cult or something like that. This is really between two people, likely practitioners, who are in exchange or who met, and then decided to share their knowledge, often studies archaeology is very distant.  You're not distant to some people, you don't know their faces and you don't know who they are, right there. Hey dude, hope you like it. All of a sudden, like in the middle of your invocation to Toth or whatever or whoever you're talking, you know, it's just some guy like, hey, I got this really deep cut you might be interested in. With the ritual manuals or the magic books, you basically have the handbooks, the instructions of how to create, for example, artifacts or textual artifacts, inscribed artifacts. And we do have on the other side, thousands of magical artifacts in the Archaeological Record. I wrote my PhD about the making and use of ancient magical artifacts as they are described in the Egyptian, Greek, magic or papyri. So when I studied the instructions, the artifacts described in there, in the manuals, how different they are compared to the Archaeological Record, but different in terms of, in so many terms actually, I have to give you a brief overview there. The Archaeological Record is like roughly between, depending on what kind of artifacts you count with. So if you count the demon boats too, we're dealing with between eight and 10,000 artifacts from the time span of the Greek magical papyri. So yeah, it's still difficult because the largest group are the magical gems. There are numbers estimated between about three and a half and 5,000, but it's still difficult and it's still a huge topic actually in research, how many are actually ancient and how many are cost antique. It can be that not as many as 5,000 magical gems are actually ancient. On the other side, we do have other ancient sources that haven't been really researched by now in detail in terms of the textual artifacts in there, like the ancient Greek lithica. This is just a rough estimation, but if we go there, we can see that the Archaeological Record, the largest group of magical artifacts is definitely gems with around three and a half to 5,000, followed by the cursed tablets between I think currently the Greek and Latin together about 1800. For a long time, there's nothing, and then we have about a bit over 160 currently lamelles. So that's gold, silver, tin, basically tablets, inscribed smaller tablets, and a papyri about the same amount. Basically, we have gemstones, lead, and then is papyrus and precious metals. Precious metals. Precious, precious. Okay. What we have in the ritual manuals is very different. We have descriptions for about 300 textual artifacts in there, and 268 were good enough that you have more or less the whole procedure, materiality, or the purpose, or the inscription. Most artifacts are made of papyrus, followed by tin, and then linen and laurel leaves. So very different from the archeological record. What we do have, for example, in the archeological record, with all the lead tablets, there are a lot of, the vast majority is actually connected to erotic and love. And a lot of spells of attraction are written down, or we know as cursed tablets are labored as cursed tablets. But the ones, the lead tablets we do have in the ritual manuals, they are not actually addressing this kind of attraction. They deal with love and erotic, but not with this, you know, drawing a woman or a person towards you. I think we have 10 or 12 lead tablets that are described in the ritual manuals compared to like 1800 in reality. You can't say that 12 and 1800, I don't compare the numbers directly, but I mean like, from all the 300 texture artifacts that are preserved, only 12 are made of lead. So this is a very small group, and in the archeological record, the lead tablets are the second largest group. So this is really different. I wonder if that has to do with that all the magical papyrus we have are actually from Egypt, but the cursed tablets and the gems, of course, we find them all over the Roman Empire. So you have them from Great Britain down to Egypt, then from Western Africa and Spain to Crimea and further to the east. So if this is because the manuals are all from Egypt, actually we don't have many cursed tablets from Egypt, maybe that specific preferences of clients were very different in Egypt compared to other Roman provinces. Another thing that's really interesting and a difference between the archeological record and the Greek magic papyri is the purpose of the artifacts or the purpose in general that we can see in the rituals and in the artifacts. The purpose that occurs most of the time, so that's protection. It's amulets, and you know, we should think, okay, that's, you know, we have a lot of amulets in the archeological record. Many gems do not explicitly say, protect me or protect the wearer, but we assume that they were used for protection. And in the ritual manuals we learned that the vast majority of the amulets was created for the magician to protect the magician during the practice. There is not a lot of amulets, very few amulets that were made for the client and then usually in context of an exorcism, you know, when there was an exorcism and then the client gets an amulet to protect him that he doesn't get possessed again. Most of the protective artifacts are actually for the practitioners themselves. And then really interesting questions and most of the instructions do not provide us with an answer is, could you reuse an amulet?  But when the ritual is over, do you have to craft a new artifact when you perform the ritual again or can you use it? When the amulet is made of gold, you can assume that it can be reused, but we still don't know if you have to do like special prayers or invocations over this artifact. You know, you speak over the artifact. Do you have to repeat this? And especially when you have artifacts, protective artifacts that are made from organic material, can you reuse them? And then next question, if you can't reuse them and with some it's clear in some cases the ritual says it's one time performance kind of. What do you do with the artifacts? I mean, you know, they're usually inscribed with the names of higher powers and they were created in a context of ritual manual in which higher powers were involved. Can you just throw them away?  The artifacts that can be reused might be for a certain clientele versus, you know, one use artifacts, you know, something to explore there, I'm sure. All right. You know, you actually, it doesn't just apply to the artifacts that the practitioner crafts for himself or herself, but also for the clients. That's true. I haven't heard about that. Yeah. And it's very interesting just hearing that these amulets were mostly for practitioners. It kind of reminds me of that whole period of time when the teachers getting back to Pidea for a second, like the teachers would often accuse each other of sorcery and like binding their tongue, each other's tongues, because, you know, being a rhetorical master was very important. And, you know, we have lots of these reports of like them trying to protect themselves from the sorcery of another rival. What we do have in terms of the archaeological records, it really seems that Egypt is the only game in town just by nature of the climate and how it preserves the papyrus versus other areas. So it really, it might be a skewed record, but we have to take in, we always have to take that into consideration that just due to climate, Egypt is going to be where the majority of these discoveries are going to be made. My experience is that there's a lot of people who actually are not really aware in terms of, because you don't think about it, if you don't have to, that actually all the magical papyrus, we have like the Greek magical papyri and the Egyptian magical papyrus, they originate all from Egypt. There is no magical papyrus from, and we do have papyrus from Italy, but there is no magical papyrus so far, or so far I know of, from Italy. They are also, the published ones in Paisenanism Bets, they are all from Egypt. And when you read studies and research about the engine magical papyri, it always seems so, you know, especially the Greek magic of a papyrus, it's so Greek. And you think, well, it's maybe, maybe it is not all that fascinating or mysterious that there are so many Egyptian terms in there, not only gods, but, but language, when you consider that it's actually, they are from Egypt, you know, even if that's, if it's Greeks that, that wrote them, they still lived in Egypt and it's very likely that, you know, they, they heard Egypt, the Egyptian language, basically every day and they saw the Egyptian temples and monuments every day and hieroglyphs, the inscriptions every day, they were all around them. It's sometimes that I myself have to have to remind me and say, well, yeah, they are from Egypt, from all over Egypt, you know, it can be from some Egypt as well, you know, and not only Alexandria, everybody who is really interesting in the study of engine magic, don't study only the papyri. It's, it's, it's, it's handy because you have the translations, but take a look at as many archaeological artifacts as well, because they literally are from all over Europe. At the beginning of scholarship, it was like, oh my God, look here, we haven't, we have an inscribed lamella we found in Great Britain and it's inscribed in, in, in Hebrew. Oh, wow.  They traveled a lot and of course they, they transported a lot of information and a lot of knowledge and traditions, not only people and, and food and, and trade in terms of material trade, but also the immaterial goods that, that, that were traded. If you're interested in engine magic, really, really look at the artifacts and most of the artifacts we have, we know that, that they are magic is because they are inscribed. So read them. It's, it's awesome. You get a much better picture. When you study engine magic, everyone is all the Greek magical papyri, the Greek magical papyri. And yeah, it's, it's the best known body of texts. But actually, it's just a very, even as Egypt, but it's very local, very small source for the study in ancient magic. And once you, you don't only know that, but you, you've become familiar with the, with the artifacts. You know, you, it's like this, you're in a house with a lot of locked windows and doors. And the more you learn, the more you see from the ancient source, from primary sources, actually, it's like each artifact opens a window.  And it's not this, it's not just the magic of papyri, really important and, and tremendous source because, because of the manuals. But still it's just, it's really basically a small, a small slice of cake. The archaeological record, just looking at it, aside from the manuals that we find in Egypt, it really fills in a bigger picture, like you were saying, it's like having one piece of the puzzle versus the whole picture. And, you know, you can make a, an analogy to when somebody's reconstructing like a, an authoritative text. Dr. Dr. Suiza, if you could tell us a little bit about your Patreon and your book projects. Thank you for giving me the chance to, to introduce my, my new Patreon to your audience or to our audience. It's ancient magic. It's patreon.com ancient magic. You can support me on, on Patreon by in, in sharing my research, in doing the research and in sharing the research with a broad audience. Be the videos, you can become a video patron, you can become a magic science patron, help me, especially support me, especially in research and ancient magic science. Or you can become a creator patron and you kind of support all the stuff I'm doing and sharing. Be the videos, be it on Twitter or on, on, on Instagram. And I'd love to see you there. Yes, everyone go check out Dr. Suiza on Patreon, check out her projects, check out her YouTube channel. Thank you so much, Kristen. This has been fascinating, brilliant, as always. I hope to talk to you soon. Until next time. Thank you. You're welcome.


I'm not 415 as my computer's telling me. So we're gonna get started. My name is Erica Fike. I'm the conference organizer and I'll be the chair for this package demo. Just for a few things, this is the first package demo of this conference. If you're here in the room, if you have a question, please raise your hand. Niles has asked that he be interrupted with questions, not wait till the end. So feel free to raise your hand and we will bring a mic over to you to ask your question. Online, same thing. You can raise your hand in Webex or you can put your question in the chat and we will try and get those questions over to the speaker as soon as possible. And then we will have a Q and A at the end as well. Yep, those are all the housekeeping things I needed to say. So let me introduce you to our speaker. This is Niles Elling. He's a postdoc at the University of Zurich and he's presenting a framework for multiplexed image processing and spatial analysis. Perfect, yeah. Thanks for the introduction and thank you all for joining. I also want to thank the organizers and everyone giving technical support here. It's really great, makes it really easy. Yeah, so I'll be looking at the chat once in a while. There will be some time where I take a little break and ask if anyone has a question so we can discuss in between. So here you see the landing page or like the compiled HTML of the workshop that I'll be giving today. Most of the demonstration will be an R but you can follow along. There should also be a link in the program to this HTML. So what I'll be presenting is a relatively newly developed framework for multiplexed image processing and spatial analysis. It is made up by three packages. One is called Steinbock. It's a Python-based, dockerized framework for image handling and segmentation. And most of the workshop will package demo will be on IMCR tools, which is a package to read in, especially annotated single cell data and to perform analysis on this. I will also quickly demo Cytomap again. This is a bit of an older package to visualize composite images and segmentation masks. So yeah, the code that I'll be presenting today is on top. So when you click on this link, you will directly come to the GitHub repository where you see this R Markdown file here. So you can test it out. I also provide the packages that you need to install and instruction how to get this script. So I want to start with a general overview on the tools that I'll be discussing. So we start from multiplexed imaging raw data. Ideally, these are just multi-channel TIF files. Here in Zurich, we're working with imaging mass cytometry, which is a commercialized system. And they provide raw data in the so-called MCD format. So there's a small pre-processing step that Steinbock can handle to generate these multi-channel TIF files. Steinbock is a framework to read and TIF files to perform segmentation by using a pre-trained neural network. It can then also quantify single cell features of these segmented cells and export spatially annotated single cell measurements, as well as segmentation masks and multi-channel images. MCR tools can directly read in the single cell data to then perform a bit of spatial analysis and spatial visualization. And CytoMapper can read in multi-channel images and segmentation masks for visualization. So I'm not gonna demo Steinbock here. For this, you would need to install Docker. I provide instructions here where you can download raw data if you want to test it. The folder, the raw data will be downloaded in the data folder. In Steinbock, there's a raw data folder that contains four zip files, which contain the raw data, and a panel file. This panel file is only there for annotating the individual channels that were acquired. So in our technologies, we can acquire up to 40 proteins at once. We use metal text to label the antibodies. So this panel file just indicates which antibody, for example here, this is SMA, was labeled with Indium 115. There's also one column indicating if this channel can be used for single cell segmentation. For example, nuclear for nuclear segmentation and then for cytoplasmic segmentation. The Steinbock framework is, well, initially, it's a Python package to make it user-friendly. It's dockerized. So what you need to do is you can set an alias. So the comments that I'm showing here, these are just bash commands. You can set an alias to Steinbock, which is just a docker-run command. Here you need to adjust the folder where the raw data is stored. And at the end, you just need to specify which Steinbock version you're using. And then within these six commands, you can read in multi-channel images. You can filter them. In this line, you segment them using deep cell. This is a pre-train neural network image segmentation. And then you can measure different features of the segmented single cells. The first one here is the mean pixel intensity per channel and per cell. These ones are morphological features as well as the location of individual cells. And the last one is neighbor detection. So here spatial object graphs are constructed indicating if cells are in close physical proximity. And all of this will be written out in a folder structure. I can also show you this here. There's this image folder that contains the multi-channel TIFF files. So here, while looking at the raw data, we have four patients. But patient one actually has three images acquired. Patient two, we acquired four images. Same for the masks. We get each multi-channel image, one segmentation mask. After measurement, we also get the intensity measures per image as well as the morphological features and then the neighborhood graphs. I can also quickly show you how these multi-channel images look like by just dropping them into Fiji and then just adjusting contrast and brightness. So here we have 40 channels, so 40 different proteins measured. This one, for example, here is Ekaterin, a tumor marker. And the last ones here are nuclear markers. And in the mask folder, you have the matching segmentation masks. This is segmentation mask for the first image. So you can see that roughly the structure fits. Here, every round spot is a single cell with an integer ID. But I will come back to this later. So this is what the Steinberg framework writes out. Due to this very standardized form, it's pretty easy to read in this data into R. To make it easier for you, I also provide this data on Zenodo with instructions how to download it. And the real package demo will basically start now. So we'll switch to R. And for the first sections, I will demonstrate how to read in specially annotated data using the IMCR tools package. So here IMCR tools provides the read Steinberg function, which reads in the generated data from Steinberg into a spatial experiment object. So here we have a spatial experiment object now with 46,000 cells. The counts as say stores the mean pixel intensity per cell and per marker. So here cells are in the columns, markers or measured proteins are in rows. The call data of the spatial experiment object stores cell specific metadata. Every cell has an integer ID. We also measured the area, the major axis length, the minor axis length, the eccentricity of the cell. And we also read in some metadata from the images from where these cells were derived. In the case of a spatial experiment object, the cell locations are stored in the spatial quartz slot. So here we have the X and the Y position of the cells. The spatial object graphs. So these are, this is an edge list indicating if two cells are in close physical proximity are automatically read into the call pair slot under the name neighborhood. So here we can see the first cell of the spatial experiment interacts with the 27th cell and so on. So this is important for visualization later. The row data of the spatial experiment object stores information contained in this panel file that I showed you earlier. So this is just information based on the antibodies that were used. And again, here the marker SMA was labeled with the metal in your 115. The rest is relatively irrelevant here. I also provide you a fully processed spatial experiment on Zenodo. So I'm only reading this in here now because this spatial experiment object already contains a cell phenotype. So in the last entry here, for each cell we have identified if it's a tumor cell, a T cell, a B cell. Yeah, of note, this is a cancer data set that I'm presenting here, containing different cancer types. The full processing was done in this IMC data analysis book. I've added the link here. So this gives a more detailed overview on how this analysis can be done, right? So now we have read in the single cell, especially annotated single cell information extracted by Steinbock. In the next section, we want to read in the images. These are once multi-channel images as well as segmentation masks for visualization. For this, we can use the cytomepper package. I wrote this package to handle multiple multi-channel images. It's really heavily based on the package EB image, which is a really great basis for image handling in R and treat it a bit to just handle multiple images. So here, the load images function can read in multi-channel images, as well as segmentation masks. When reading in 16-bit images, and this is how we save segmentation masks, you have to set this as is parameter to true to really read in integers. And you can also look at these individual entries. So here, this is an EB image object which stores integer IDs. And here, these are pixels that all come from the cell with the ID 57. We can now set the channel names of these images. So currently, images have no channel names. In the case of the Steinbock framework, the multi-channel images are stored in the same order as the single cell data. So we can directly transfer the row names of the spatial experiment to the multi-channel images. And now the CytoImageList object, it's a class exported by CytoMapper, contains 14 images, and each image contains 40 channels, similar to what I've shown earlier in Fiji. One important thing is also to add the element metadata of the CytoImageList object. This is important to match images between the multi-channel images and the segmentation masks. We want to make sure that the names of both images and segmentation masks match, so they're in the same order. And here we can set the element metadata of the list to contain the sample ID. So sample ID is an image identifier. This can now be found in the element metadata of images, of masks, as well as the spatial experiment object. So there's also sample ID here. So now you can link images, the segmentation mask, and the single cell information contained in the spatial experiment object. Right, so another option would all be to generate single cell data directly from the multi-channel images and the segmentation masks by using the measure object function from CytoMapper. I'm directly gonna start this because it's gonna take a couple of seconds. So here for every cell contained in a segmentation mask, it calculates the mean pixel intensity for every channel and then also records morphological features and the locations of individual cells and stores this, in this case, in a single cell experiment object, but since a couple of days, CytoMapper also supports spatial experiment objects. And at this point, I can ask if there are already questions now since this will still run for a couple of seconds. We're not seeing any hands raised in the room. Anybody? I mean, that's good. Yeah, you're being very clear. So what's happening here is now, well, the operations are quite heavy. So it needs to iterate through all images. I mean, for 14 images, it's really manageable doing this for, I think we've tested it on 700 images and that was one hour. So using Steinbord for segmentation is definitely recommended. The back end of the single cell experiment, Aya. So I mean, this is all done in memory at the moment. The single cell experiment is usually actually quite small. So we never had issues with this keeping it in memory. CytoMapper also supports images kept on disk. So they are stored as H5 files. You can either generate them in external software or you can use CytoMapper to read an images and then write them out as H5 files. It does, however, make everything slightly slower. So the memory usage is minimal, but the computations take a bit longer since individual images need to be read into memory first and then processed. So here the created single cell experiment object contains again, 46,000 cells. It doesn't, well, it has row names based on the channel names that the images had. And yeah, relatively simple call data annotation based on the morphological features. So again, the area, the radius and then the X and the Y location. Since this is a single cell experiment object, it stores in the call data and not the spatial quotes. So will Steinbord also work on single channel tips and would that have an issue with CytoMapper? Not really. So we haven't really checked Steinbord on single channel tips. In theory, it should work. There might be an issue with the segmentation because you would need to have at least one channel for the nuclear stain and one channel for the cytoplasmic stain. So ideally you want to have a two or three channel fluorescent image for segmentation. CytoMapper can handle single channel tips for measurement and also for visualization. So for visualization, we're gonna select these three images. What if you segmented outside Steinbord? Yeah, I guess, I mean, like cell posts and status, they support only nuclear segmentation. So I guess they are a single channel will be fine. We tend to do a whole cell segmentation. But yeah, if you're using cell posts, then there shouldn't be any issue and it will create a similar segmentation masks to what Steinbord is doing. Okay, so we have selected three images and we can now visualize these images. Here I'm coloring them based on six markers. And this BCG parameter stands for background, contrast, and gamma. So I'm just enhancing the contrast for each marker. We can also zoom in into these images. So here in red, this is Ekaterin. It's a tumor marker. In green, you have CD3, a T cell marker. In blue, this is CD20, a B cell marker. Turquoise, you can't really see since it overlaps. It's cytotoxic T cells, it overlaps with green. CD38 in magenta is a plasma cell marker. And K67 is a proliferation marker. So cytomapper only supports up to six colors due to spectral over, at some point you can visualize more colors. It will just be white. Let me finish with the visualization section and then I will come back to the question in the chat. So here for pixel visualization, we use the plot pixel function. Cytomapper also exports the plot cells function. And this now takes segmentation mask objects and can color each cell based on metadata or based on the expression. So in this case, we can visualize the matched segmentation mask and color them by cell type. So here up here, you can see some tumor cells. And here in pink or light purple, you see cells that we call B next to T cell due to our low resolution. We can't really differentiate between B cells and T cells if they sit really close to each other. So we just call them B cells sitting next to T cells. You can also provide custom color vectors. And in this case, the color vector is a named vector. The names here are the cell types and the colors are, or indicate how these cell types should be colored. And then you get the same segmentation mask out just a lot differently. Also one thing you can do is you can subset the spatial experiment object and you can specifically color a certain cell type of interest. So here by setting missing color to white, you color all cells that are not part of the spatial experiment object as white and all other remaining cells. In this case, CDAT cells are colored in red. Okay, so there was one question if you could write your own custom feature extraction function for a measure objects. So the measure object function supports a lot of features already, so you can calculate the mean, median, standard deviation in intensities per cell. You can calculate any sort of morphological features. It also supports all funky, hydraulic features, it's just something that EB image provides. Custom feature extraction is currently not supported but I mean, it would be possible to add this via an issue or pull request. And then, so the EB image propagate a cell body. Do you have experience with EB images propagate cell body segmentation? I've used it before, I don't remember that it does a Voronoi tessellation, but I've never used EB image really for segmentation other than also thresholding and object detection. So I can't say too much about this. Good, so in the last part of this demo, I wanna switch to spatial analysis. So working with this spatial experiment object, we have spatially annotated single cell data and we can do some relatively simple things using IMCR tools. First, I've mentioned before that Steinbock generates these spatial object graphs by expanding, so it does it by expanding the mask of every cell and then cells are considered in close spatial proximity if the expanded masks overlap. The IMCR tools package also provides the build spatial graph function. So here you can use or you can calculate these graphs are talk and using different settings. For example, you can construct a KNN graph between cell centroids and expansion graph. So here you only detect interacting cells in a certain distance or you can also construct this graph via Delaunay triangulation. We can run all of these commands. So here, yeah, we detect a 20 nearest neighbor graph and the next graph uses an expansion of 20 micrometer. And when you're using the Delaunay triangulation, it's worth setting a max distance otherwise you get a funny border effects. All of these graphs are stored in the call pair slot under different names. So here the first entry is the graph exported from Steinbock and this is the KNN interaction graph expansion and Delaunay. And now using the plot spatial function from IMCR tools you can also visualize these graphs. So here we are just selecting one image and we also want to draw edges and we color the nodes by cell type. So when we do this, so I hope you can see this. Here we can make it big. So here every dot is the centroid of a single cell. It's colored now by cell type and then you have these gray edges between interacting cells. So Steinbock generates an undirected graph. So you have these bi-directional edges between cells. The KNN interaction graph, it now contains a lot more edges because we detect the 20 nearest neighbors and it also takes a bit longer to plot since there are a lot of edges that need to be drawn. So here you can see these cells are highly connected and I wouldn't really consider them direct names anymore, but a 20 nearest neighbor graph now accounts for larger interactions and we can also see a bit further in this demo why this might be needed. And then the last one is the Deloni triangulation graph. Actually I skipped the expansion. So here Deloni triangulation, it also detects these a bit more long range interactions between cells. One feature of this plot spatial function you can also here give the image ID and it will plot all images side by side. I mean for our data, so our images are relatively small. They're only 600 micrometer in width and in height. And this doesn't render too nicely now. Well, yeah. But here you can already see different structure. For example, these four images here contain the so-called b-next-to-t cells indicating tertiary lymphoid structures. All right, so for the per cell protein expression level to use mean or total intensity of the pixels belonging to the cell. So we usually use the mean intensity which is just the total pixel intensity divided by the area of the cell. There are also other ideas of normalizing this better but for us the mean always worked. Some people also tend to use the median but if you have some markers that are only expressed on like certain spots of the cell or just on the membrane the median will be zero and this might be an issue. Good, so the next part of this demo is how to calculate cellular neighborhoods. So this is a term coined by Gary Nolan's lab and they published two papers based on this. The approach is relatively simple. So for every cell you basically calculate the fraction of cell types in its direct neighborhood and you then cluster based on these fractions. So the IMCR tools package provides the aggregate neighbors function and then in the, this function generates a new entry to the spatial experiment object that is a data frame. So every row here again is a single cell and then you have the fraction of cells in its neighboring or in its neighborhood. So here I'm using the 20 nearest neighbor graph. So I'm aggregating across the 20 nearest neighbors in 2D. And for example here for cell four, five percent of neighbors are CD4 cells 10 percent are t-rex and then the rest are tumor cells. And you can then use this information to cluster the cells. Here we are just doing simple K-means. And then we can visualize these cellular neighborhoods. So here these tertiary lymphoid structures they're part of this pink neighborhood. The cyan cells are tumor cells and these purple cells are the tumor stroma border cells. We can also visualize the composition of each cellular neighborhood as a heat map. I'm not gonna go into details here since there are only five minutes left. And for brevity, I'm also gonna skip the patch detection function. This one allows you to detect patches of predefined cells and directly move on to the interaction analysis. So this was proposed by Shapiro. It is based on a permutation approach to identify cell type pairs that interact more or less frequently compared to what you would expect by chance. Gonna start this since it also runs a couple of seconds. So what it does now is per image and per cell type pair it computes the average interaction count. And then it shuffles the labels in this time in this case 200 times and generates an empirical null distribution of the average cell type interaction count. The actual count will then be compared against this empirical null distribution and these empirical P values can be calculated.  Yeah, the default iteration parameter here is 1000. Usually it also works if you permute to 300 times or so. Good. So the result is a data frame for every image and every cell type pair. It lists the average interaction count and then provides empirical P values. And the sick well in entry indicates if two cell types are interacting just one. If there's no statistical significance which is zero or minus one meaning they avoid each other. These values can also be summed up across all images and then you get these heat maps where the color indicate if certain cell types are interacting. For example, here B cells interact with B next to T cells. Two more cells interact with each other but they avoid all other cells. So they're quite compartmentalized and CD4 T cells interact with T-Rex which is not super surprising. And with this, yeah, I'm already at the end. There are also more or there are links to all the other resources that I mentioned. And I'm not sure if we have time for questions now. I have one question in the room and I think that will be our last one before we move on. Hi, thank you for the talk. So, okay, maybe it's a bit early for me to ask this question. Okay, I'm actually going to present about it tomorrow in the package demo. So I wrote a package called spatial feature experiment which extends spatial experiment. But unlike your package, it stores the cell segmentation as vector polygons with the vertex coordinates and instead of mask, so it's more memory efficient. So if, okay, so maybe like tomorrow if you get a chance to take a look at a spatial feature experiment, like what you consider like integrating spatial feature experiment with into your package. So can reading your data as SFE in addition to SP and SCE? Oh, sure.  It would be nice. Yeah, I'm not sure if you already have a reader for converting segmentation mask into polygons but I can also check tomorrow. We're using segmentation mask to overlay outlines of cells on composite images. So you still need to store images somewhere. Yeah, so I also roll like applying functions for the polygons. And actually you can convert the masks into polygons such as using the Terra R package which is designed for raster geospatial data. Okay, cool. Yeah, that's nice. I will check this out. Any other questions from our in-person audience? Okay, one quick question. So in the interaction plot you are showing, what does T-Rack interacting with self mean? Like on the diagonal, T-Rack interacting with T-Rack, for example. So yeah, it's a common thing for our data. We usually observe certain cell types to interact with each other. This could be biological, so it could mean that T-Racks are sitting next to T-Racks. It could also be a segmentation or phenotyping issue where, for example, a single T-Rack is split into two. It's sometimes a bit hard to tell. For example, we know that B cells, they tend to cluster. So they form these large bulk of B cells, these tertiary lymphoid structures. So there we expect them to interact more often compared to a random distribution of B cells. But for example, here in neutral fields, they don't really tend to cluster except for a necrotic areas, which we didn't acquire. All right, well, in order to give everybody time to get to the next session, we're gonna close this one. Thank you very much. A round of applause for our speaker. Thank you so much. Thank you. There is a session here in just a few minutes, so you can either stay or head back to the cure building. Thank you. Bye.


To start, my first question was about last night when President Trump announced that they had a partial deal with China and what was your reaction to it? Did you think it means anything? Is it good to have a little push on the brakes? Well, I think it means something, but not much because we had this kind of examples already two or three times. Right after the G20 meeting in Argentina, we had a truce at that time, and then they met again in early March, and they talked about something, and then they go into another truce in May, same thing. So this time, they're just delaying the extra imposition of tariff after Christmas, whatever. So it means something, but we don't have any visible agreement yet. That's my observation. That's a lot of stop-and-go, as you said. What was your reaction to it, Carl? I like the fact that the parties are talking and there seems to be some convergence, but I share the view of my neighbor. It is a little bit of a relief for a very short time, but if the big goal of the US, of the president, is to shrink the deficit that they have with China, a lot more is necessary, and I think a lot more inside the US is necessary. Gabriel? Yeah, so I think economically this is pretty much insignificant because the tariffs that have already been put in place don't go away. Further escalation is avoided, and some of the things that are promised now happen anyway, so imports of pork meat, for example, into China are going up very strongly already, also from the United States because they had to slaughter large shares of the big population because of an epidemic. And on the other hand, the US president is not very eager to have tariffs on Christmas presents, so I think the pork story and the Christmas story, and then we have this sort of small deal, but it does nothing, also uncertainty, the big issue we discussed this morning is not going away. One could even say that this type of negotiation actually exacerbates uncertainty because it shows the back and forth and how transactional this all is and how much showbiz is there out. It's very much like Trump, you know, transaction showbiz, I mean we're very much into it. What about you, Marcus? Transactions and showbiz, yeah, I think these will be running themes for the next hour. One hopes that this is a first steps or something greater, but I am skeptical. Donald Trump is an avowed protectionist. He makes no bones about it. And the people who surround him do not support a liberal rules-based international order. They are not friends of the WTO. They want to see trade organized through bilateral managed trade deals. If they resemble anything, it's the European governments of the 1930s. And Donald Trump, as you mentioned, launched a series of trade wars. It is important to understand how big this is in the US context. If you simply calculate the implied, applied tariff rates on either imports from China or imports from the world as a whole, Donald Trump has basically moved the United States from the neighborhood of the EU or Japan with applied rates of less than 2%. To the neighborhood of Brazil and India. We're going to be another big emerging market with populist political leadership. The implied, applied rates on China would be more than 25%. There's huge numbers. So if you look at the specific deal last night, first of all, there's no deal. There's no text. There's not even a joint statement. The Trump administration made assertions that have not been repeated in China. So we don't know what that's about. There is a postponement of tariffs. The tariffs were supposed to go from 25% to 30%. That's been postponed. But it's not been taken off the table. And the core issues that supposedly were the justification for the trade war in the first place, intellectual property rights, forced technology transfer and so on, are not talked about. So one hopes this is the seed of something that becomes much greater, but it doesn't look like much. One last thing about show biz. This also has to be understood in the context of the impeachment effort underway in the United States right now. Trump's support among the farmers is eroding. He has been hurt badly by the Chinese retaliation on soybeans, as well as not only on pork from China, but the fact that he pulled the United States out of TPP. So the United States producers faced big trade diversion problems in the Japanese market on pork. So an audience like this needs to expect over the next year or so, not just in the trade area, but foreign policy area, that as the impeachment process ramps up, President Trump is going to be increasingly desperate to change the conversation in the United States and have some sort of positive or some sort of victory he can point to. And so he's going to take something like this, which appears to be nothing, and turn it into, you know, we're making progress with China, and the rest of the country is here as well. Expect comments on trade and other foreign policy issues that may bear no resemblance to reality that are being driven by domestic political messaging. Okay, so I guess we all agree that last night we have a French expression for that. It's called la poudre aux yeux. I don't know how you say it in English, but it sounds well in French, just for the show.


All right, good afternoon. Sorry you have to see my face again. Lot of people have been complaining, why is that you have so many sessions today? Unfortunately, when you organize conference and speakers drop out, you end up becoming the bakra who has to fill in all these lots. But I'm glad to see so many people interested in this topic. And I know that this topic is very interesting for a lot of people. Like many people came up and said, we're really looking forward to it. So if you're really looking for golden answers, I would like to set the context right now. You might not get those golden answers, right? What I'm going to give you is some techniques that I have found useful in working over the years in this approach. And maybe that will give you some food for thought that you can go back and start applying in your context. But please adapt to your context and try and see how it works in your context.  And I'll try and balance it with some of the concepts that I cover and get you guys to do some things, right? So we'll be working in groups.  Try and find a table where you like people around you. I would encourage if you guys can come in towards this side. So it'll help other people who come in later to come into this session. And people who want to run screaming, it's easier to go out. I didn't know what kind of an audience to expect. So I did have a little placeholder over there to really just touch upon user stories from my perspective what user stories are. And then kind of start diving into various different techniques you can use at a high level to actually break large stories. And then only focus on splitting as one of the techniques in which we will go and then we will talk about various different techniques within splitting of how you can split the stories, right? But there are other things you could do, not just splitting stories, you could do other techniques which we'll talk about. And then some high level stories in terms of a high level prioritization approach itself which helps you better stories, right? So that's the agenda, one and a half hours, very ambitious. Last time I ran it, we overran by two hours. So we will see how we go. But let's see. We're gonna start with activity where I'm gonna quickly distribute example. And I want you to form groups.  How many people here are familiar with the, as I want to show that format? Hopefully by the time you leave this room, you should forget that format. You know who came up with that format? Mike Cohen, sorry though. He's popular for taking other people's work and bringing it to people, which is good thing. But it was not him.


The Student Loan Forgiveness Plan has been canceled by the Supreme Court. Over 40 million loan borrowers are in jeopardy. Those who have to make payments must resume them this October. Have you defaulted on your student loans? Is your debt in collections? Are you considering filing for bankruptcy? Or have you been defrauded by your college? Purchase your legal plan and get the advice and consultation you need. Visit www.inivery.weerlegalshield.com