The reason the neurons all died after the initial seeding is because you didn't rinse the PEI off properly and/or didn't dry it properly. While PEI is the best pre-coating agent in the business, it is also an extremely potent membrane perforant when in solution, so any miniscule picogram of the stuff you left unpolymerized is gonna be jumping from cell to cell, shredding their membranes. Rinse at least five times with maximal volume your growth vessel can take, and dry the vessel overnight in the LAF bench, otherwise you're just wasting expensive reagents. Source: neuro lab-slave for 10+ years
Don't you just hate it when you're chilling in the torment nexus when all of a sudden your entire being is overcome with an extremely powerful chemical irritant?
Me: Wait, where does the "torment" come from in Torment Nexus? The Thought Emporium: Cayenne pepper flakes. Look at the neurons screaming in torment. Delightful. Me: Oh yeah, that makes total sense, thanks.
@@jakobweisser8044 "We built a neural interface on an unregistered cruise liner off the coast of Panama that mines cryptocurrency. We just need your donation of $200,000 to keep it running and to pay the illegal waste dumping fees." -techbro neuroscientists
9:55 Aha! apropos to my previous comment about cell density, I see how to solve your problems: We always added a small droplet (50-100ul) of cell suspension added to an EMPTY dish, with just a 3mm laminin wet spot. Calculate what density you need in the suspension to get 50k-100k cells in that droplet. After the cells have settled and adhered (30 min?) carefully add the rest of the medium to flood the dish. Now you will have very dense cultures and signals on every electrode.
@@mihael64 My advisor literally can not physically stop himself from talking about Dune if he drank more than 2 beers... And not every researcher is crazy. Some can even manage work life balance. Not anyone I know, but I heard some can!
I'm a doctoral researcher and Neuroscientist. When it comes to culturing medium for neurons morphology can tell you a lot about the state a cell is in (this goes for most cell types). To me the classic choice of Neurobasal gives you cells with shrunk cell bodies (the nucleus is tightly shrink wrapped if you will) and long complex truncated neurites (dendrites and axons). This tells you that these cells are "mature" neurons. They are not quite mature because you literally need let time pass for them to become mature, interestingly the time of maturation depends on species and is much longer for human neurons compared to mouse neurons.
@@TheAmazingAnimator it is more about complexity of their connections and morphology and less about the overall number of cells. In culture you can observe the same time discrepancies between neurons of different species, so it is an inherent quality of the cells. We are not clear on what level of consciousness other animals have , since we can’t test for it. You probably mean animals become independent much earlier, since humans literally complete their development outside the mothers body. During evolution our heads got to big for our pelvises so we have to be born “prematurely”.
I loved reading "Don't Build the Torment Nexus!" When I was a kid. I don't remember what the moral of the story was, but I really liked the lore, aesthetics, and world building. I'm so glad someone is bringing my favourite sci-fi to life! We really do live in the future!
@@Penguinman2.0Neither can I… On a second reading, I don’t think it’s genuine, “I don’t remember what the moral of the story was” seems like it wouldn’t be there if it was.
@@Penguinman2.0 The book doesn't exist, its from a meme talking about how techbros proudly claim inspiration from scifi stories where the whole moral is NOT to build the technology featured in the movie. Like how vr/metaverse nerds constantly say "its like the Matrix in real life"
@@SissypheanCatboyThe first thing we did after GPT-1 showed promise was hook up the next version to the internet to learn from. It's like the people at the forefront of advancement have turned dystopian prophecies into a checklist..
Guys, don't stop. I NEED to see what will come out. Your first 2 vids inspired me so that from just a noob in programming thinking to himself "well, im gonna be a programmer and program commercial things" you made it so I now have a greater purpose in life and right now I'm doing an EEG research together with a neurobiology lab (I was the one who contacted with them and suggested a research and I'm the one doing it, but they give me access to some fancy things). Thank you and DON'T STOP UNTIL ITS DONE
Hey! I work with mammalian cell cultures, and I know they can be very finicky! If you didn't change the media a day after your initial seeding, the residual DMSO from the cryopreservation media is likely what killed your cells. Not sure if you did this or not, but I didn't see it in the video so figured it was worth mentioning! You guys are awesome, keep up the great work.
My other suggestions seem to have been deleted, perhaps because I linked a paper of ours. I guessed that you may be killing cells with too much light. Phototoxicity is a huge problem with any fluorescence microscopy. To reduce it, use phase contrast (not fluorescence) microscopy and use longer-wavelength (red) light. Also, when doing fluorescence imaging, use a lower-NA lens, more sensitive cameras, dimmer illumination, longer wavelength excitation, and shorter exposure to light.
Feel free to send me an email with all of the suggestions so they don't get lost! And they're greatly appreciated as always. We only use phase contrast when checking on the cells. Fluorescence was only used at the end for the sake of the experiment, and the dye was only used on one of the arrays after we had finished all other experiments. The arrays are also only ever left on the microscope for as brief a moment as possible, but the phase contrast images aren't as pretty so I don't use that footage in the video as much. I also saw you recommend concentrating the neurons in the middle. I read that in the instructions and that's how we attempted it the first time when the cells died. We went with adding the neurons after the media because we knew for sure the neurons would survive as we've tested that method a lot and couldn't risk another culture dying. We were pressed for time to get the video out. And for our arrays the electrodes are REALLY spread out, so we're gonna need a high neuron density across most of the surface of the array anyway. You'll notice I made the walls of our arrays taller than the multichannel ones. It was specifically because I wanted to increase the amount of growth media per well so the neurons don't deplete it as quickly and to account for an eventual much higher neuron density across the array. We have wanted to upgrade our camera on the microscope for a while, but the scope itself was a HUGE upgrade. It's new for this video and it cost a pretty penny. But it's built exactly for this kind of experiment so we can switch out the camera next. I also want to move to a red dye to lower the illumination wavelength as you say. And also just to make the videos look more interesting than always green. We ran out of time, but we also got a tubulin specific dye we're gonna show off next time that would look best against an otherwise red stained neuron. And we picked up NucBlue so we can stain the nucleus properly. One of the shots of the neurons firing included it from the first well we tested it on. I overlaid it. It's subtle but makes the neurons look nicer and the nucleus visible.
@@thethoughtemporium youtube doesn't care and if the automated system even imagines your comment as bad it will just automatically delete it or hide it
Just want you to know that your channel is what introduced me to the field of biological engineering. I'm now pursuing a doctorate in the field. Thanks for being an inspiration.
For the ones randomly watching this and has no idea who Steve Potter is and why it is a big deal It is as if Jesus Christ himself showed up for a theology class
I had no idea you could actually grow live neurons and train them to do stuff... thank you for documenting your unique experiment and sharing it with the world.
This part of RUclips is interesting because you have genuine, field respected scientists contributing to a meme. A similar situation I see is a video series an amateur arachnid enthusiast making absurdly detailed videos about common house spiders, and basically has tons of scientists conversing with him and in the comments about the most trivial of differences between two species of harmless spiders
Hey! I’m messing with neurons at the moment in my PhD and I saw the exact same morphology in DMEM/F12, I was stumped what was causing it! As a precaution we’re genotyping with ICC and qPCR since we’re deriving from iPSCs - anyway, love the video! This project is so cool!
I am a mitochondrial and cancer physiologist, and not a neurobiologist, but have been studying metabolic and growth effects of different commercial medias. It might be worth keeping in mind that the metabolite concentrations in DMEM are largely not found in the brain. I'd be curious to know how Neurobasal and DMEM compare in their composition. All of these considerations, i.e., metabolic activity and ionic concentration, will affect the neuron's membrane potential.
DMEM with FBS (foetal bovine serum) is an excellent medium for accelerated growth but yeah, it's 4x stronger than standard EMEM. EMEM or Opti-MEM may be a better option for maintaining mature cultures, with Opti having the added benefit of requiring much less FBS.
The trick to finding a comfy spot for your tongue is actually the roof of your mouth. People struggle with it because it's not intuitive that you keep it up there with suction normally.
"What if we grew a mind in a vat that only knew how to interact with the world through violence and was trained to snap headshot imps in .03 seconds, this will surely not go terribly wrong."
Yeah, this could never go wrong, but to make sure lets wire a 100 of these up to the Boston dynamics robot wielding a shotgun. But we would still be in control, right? lol
I know this is a joke, but the interaction that the neurons see is not in any way related to violence. it's like if you played doom, but instead of a screen, you just see an array of 10 blinking lights and no-one told you that you're playing doom, only if you're succeeding at pressing the correct buttons or not. so ... as long as no-one tries to apply this to real-life weapons, there's no reason whatsoever to worry. though ... now that I think of it, knowing humans, this does indeed have me worried
Context- For it to work like the joke intended it would need some reason for violence like a reward system but for that to work we have to have some sort of detection software supporting it and at that point just use software, so yeah the joke falls apart, oh well. Except for machine learning being applicable to organisms, i.e. stimuli training, so maybe we could consider culturing them with positive stimuli for shooting the target before sending them but that sounds like a lot of work. Not that someone wouldn't try? Also if the logic reply wasn't to the Boston Dynamics bot joke then that is really embarrassing lol
@@DrippyPootis on the other hand, the neurons not being aware of it being violence points to another issue, since that means they won't have any concerns whatsoever with performing violence if put in a situation where they might accidentally do so. putting any kind of neural network, artificial or biological, in charge of dangerous equipment means that unless you very actively train it to avoid harming humans, it will do so if any humans get in the way. this is of course already the case with any kind of automated machines, even without neural networks or similar, which is why we tend to place robots in cages, so we can better protect humans from them
I think we are not appreciating the work they are doing: THEY ARE LITERALLY BUILDING A CPU FROM SCRATCH. Something that costs companies with billions of dollars and hundreds of engineers, but on top of that they are doing it with neurons. I mean MY HEAD EXPLODES THAT THEY DO THIS AND SHARE IT. Seriously, thank you very much, I have watched the videos in this series dozens of times and I am obsessed with this technology, you inspire me. THANK YOU!
if youre still having issues with the array electrodes being cut correctly, maybe ask a glitter company if they can help. I know its a weird idea but glitterX is just a company with precision cutting instruments. Might be worth it
@@harriehausenman8623 Yes, and it is most likely pearlescent paint for hobby boats. Boat manufacturers don't want it to be associated with how "unmanly" glitter is seen as, as well as the fact that pretty much all of it ends up as pollution in water.
Impressive! It’s really nice to see the path your channel has taken. From the time I was watching you hunting VHF satellites in Brazil, Wi-Fi imaging, the pulsar corner reflector (that one got me really excited), to DNA hacking and neuron training. I would like to say I came for the RF and stayed for the bio. 10/10
Hey... I've noticed sometimes when pipetting, you (or whoever is doing the pipetting) touches the destination, and they go back to the source. This is a no-no as it can cross contaminate the whole batch, and the source material. I'm not even talking about living contamination. In the examples I saw, I don't think it's that big of a deal, it would introduce neurobasal into the neuron source. But it's a bad practice either way.
Yep, once it touches a new source, tips go bye bye. I'm surprised at some of the stuff done here, as we ALWAYS filter our media... And I haven't seen that. Also, just pouring tubes out can be a source of contamination.
This is probably the coolest experiment I have ever seen - I am really looking forward to the next part! Another interesting aspect you could add to this would be to have two separate cultures of neurons acting on the same system. This would allow for demonstrating coordination and collaboration between independent cultures for instance.
ON TODAYS EPISODE OF MANMADE HORRORS WITHIN COMPRENTION (edit: i will respect english spelling once its concistant (was that wrong, probably) fake ass language)
For your cap problem. Get a smaller radius plug to fit in the collection. Use the semipermiable membrane as the gasket layer. You can then place an o ring directly on the plates acrylic pipe.
2:20 That tech tree reminds me a lot of Dr. Stone. I realise it's also a common enough trope that it could have come from anywhere, but the specific style of this one looks distinctly inspired by Dr Stone's approach to the idea.
2:18 "don't build the tormentor nexus. And thus far we've been making great progress"... wait wait, so, you are making great progress in NOT BUILDING the tormentor nexus, or in building it?
It's funny because a difference between a video game and a live camera feed are the same to this. I bet someone is already leaps ahead doing what you are doing but you are absolutely on the right track.
If you'd like to support our work and get yourself an array poster, check out our store:
thethoughtemporium.ca/products/uv-glow-poster
maybe
Did I get "printed locally here in Quebec" right? :o
@@Alceste_ yes, the link to the screen print studio is in the infobox
*THE PART TWO WE HAVE ALL BEEN WAITING FOR* 🎉🎉🎉🎉🎉
Hi, i think the link isn't working
The reason the neurons all died after the initial seeding is because you didn't rinse the PEI off properly and/or didn't dry it properly. While PEI is the best pre-coating agent in the business, it is also an extremely potent membrane perforant when in solution, so any miniscule picogram of the stuff you left unpolymerized is gonna be jumping from cell to cell, shredding their membranes. Rinse at least five times with maximal volume your growth vessel can take, and dry the vessel overnight in the LAF bench, otherwise you're just wasting expensive reagents.
Source: neuro lab-slave for 10+ years
Bump and like for visibility
Sounds like you are an ideal buddy for this dude to keep track of details, if you didnt have a job already.
Up!
I was about to say that exact some thing. However, you left out the part about the goop in the gloopinator. You really need to check that too.
post doc in a university?
Imagine being conscious just to find out your purpose is to do a playthrough of doom
Somehow that doesn't seem that bad, it's a decent life purpose
That's more or less how I remember my high school years...
A fulfilling life
you know that there are theories that we live in a game. Soooo
That's a lot better of a purpose than to pass the butter.
Don't you just hate it when you're chilling in the torment nexus when all of a sudden your entire being is overcome with an extremely powerful chemical irritant?
That moment when you become the Carolina Reaper lol
I'm glad someone's finally getting around to building the torment Nexus
Well it wasn't going to build itself.
@@ayybe7894 your not wrong lmao
Me: Wait, where does the "torment" come from in Torment Nexus?
The Thought Emporium: Cayenne pepper flakes. Look at the neurons screaming in torment. Delightful.
Me: Oh yeah, that makes total sense, thanks.
I'm just glad it's the humble mad scientist and not the techbros.
@@jakobweisser8044 "We built a neural interface on an unregistered cruise liner off the coast of Panama that mines cryptocurrency. We just need your donation of $200,000 to keep it running and to pay the illegal waste dumping fees." -techbro neuroscientists
Painstakingly growing a brain slice only to pepperspray it moments later -.-
Such is the cruel nature of life
Do not build the Torment Nexus. But if you do, do not torment the Torment Nexus with cayenne pepper flakes.
Sounds like a middle-age crisis
Almost human
every time i found one of his videos i get this harlan ellison vipes and i dont know why.
9:55 Aha! apropos to my previous comment about cell density, I see how to solve your problems: We always added a small droplet (50-100ul) of cell suspension added to an EMPTY dish, with just a 3mm laminin wet spot. Calculate what density you need in the suspension to get 50k-100k cells in that droplet. After the cells have settled and adhered (30 min?) carefully add the rest of the medium to flood the dish. Now you will have very dense cultures and signals on every electrode.
Thank you. We really appreciate all your input.
Yo ur THE STEVE POTTERS, NICE TO MEET U😅
"We're naming it: The Torment Nexus, after the hit book, Don't Build The Torment Nexus."
Bro is literally a classic evil scientist at this point
I cant stop laighting at that !
@@rubikscuber9277 Absolutely! (in the classical meaning of the word, like in "The Gay Falcon" (1941)) 😄
@@harriehausenman8623 sorry I mean to say Yay
@@spencerharrison9680 *tech company
"Don't build the torment nexus"
YOU HAD ONE INSTRUCTION.
Instructions unclear, built torment nexus
@@omn1cr0ngaming66 Instructions were not unclear dummy :>
It's definitely a spider mastermind. He's building a freaking spider mastermind that should never be built and naming it the Torment Nexus!
3:20 "You merely adopted the sports drink. I was *_B O R N_* in it, *_M O L D E D_* by it"
Nice constantine reference
That sounds like something Caligula would say.
@paulmichaelfreedman8334 my brother in christ, that's a batman reference
@@X-SPONGED oh.
Imagine being the neurons
"okay, so we can't sense anything, and we probably can't do anything, but we AAH SPICY!"
Imagine just being born to not see, and then you just get random feelings all around you
@@in4init3vrimagine being a worm
@@in4init3vr
Imagine being blind
@@in4init3vrthat's...that's blindness.
@@in4init3vr i dont think they feel i mean they dont have a nervous system or anything right?
"It's designed for researchers, not a bunch of crazy nerds"
"Corporate needs you to spot the difference between these images"
Researchers aren't nerds, they're just crazy
budget
@@mihael64 But... they are also usually nerds XD
@@mihael64 My advisor literally can not physically stop himself from talking about Dune if he drank more than 2 beers... And not every researcher is crazy. Some can even manage work life balance. Not anyone I know, but I heard some can!
The only difference between a researcher and a nerd is that researchers are paid.
"Don't build the torment nexus."
"What'd he say?"
"I don't know. I think he said build the torment nexus."
3:50 "long term, i want to simulate a whole brain"
fucking WHAT
Bro is a mad scientist 😭
Naturally. 🧐
I think he's not talking about a human brain (so far...).
@@samuelhomberg9075You want skynet? That’s how you get skynet.
@@samuelhomberg9075give him time...
"What is my purpose?"
"You play DOOM."
"RIP AND TEAR! RIP AND TEAR! RIP AND TEAR!"
Rip And Tear, or R.A.T. for short.
Until it's done!
Man is gonna build a murder hungry superintelligence
"Can living neurons play?", the living neurons asked while playing.
and the room of higher beings laughed.
Can't wait till I have to feed my computer and overclock it with some spicy red pepper
*losing in valorant* MY FRAMES! GET THE PEPPER
"Hey guys, my GPU is struggling to hit 120fps. Gotta go toss a 5 hour energy into the growth medium circulator."
Sprinkling it in like fish food into a brain in a tank
*playing fps game*
*loosing connection*
"Guys, my machine just had a seizure."
What i learnt is that injecting pepper into the brain will be an effective form of torture.
"Slightly sentient enchilada" caught me off guard xDDDD
That was my favourite one!! 🤣
18:05 for those looking for the funny line
I'm a doctoral researcher and Neuroscientist. When it comes to culturing medium for neurons morphology can tell you a lot about the state a cell is in (this goes for most cell types). To me the classic choice of Neurobasal gives you cells with shrunk cell bodies (the nucleus is tightly shrink wrapped if you will) and long complex truncated neurites (dendrites and axons). This tells you that these cells are "mature" neurons. They are not quite mature because you literally need let time pass for them to become mature, interestingly the time of maturation depends on species and is much longer for human neurons compared to mouse neurons.
I think that's because rats have less neurons to mature overrall.
also because other animals (I think) gain consciousness faster
@@TheAmazingAnimator it is more about complexity of their connections and morphology and less about the overall number of cells. In culture you can observe the same time discrepancies between neurons of different species, so it is an inherent quality of the cells.
We are not clear on what level of consciousness other animals have , since we can’t test for it. You probably mean animals become independent much earlier, since humans literally complete their development outside the mothers body. During evolution our heads got to big for our pelvises so we have to be born “prematurely”.
Finally! Man-made horrors WITHIN my comprehension!
😂😂😂
It's brilliant but a little sad to think that a dish full of rat neurons could potentially beat me at Doom.
the ultimate cheat system, full automation cheats; I could grind war thunder off a chunk of rat brain
@@breadboi3837imagine getting banned for rat brain botting 😂
@@zachrowe6271 🤗
To be fair, you also have to think about, yknow, having a body and pressing keys and shit.
@@user-burnerTrue that, all about focus, reaching flow state.
0:01 wtf put it back
Lol
Lmao
Rofl
It took me 2 seconds to understand lol
Lmfao
I loved reading "Don't Build the Torment Nexus!" When I was a kid. I don't remember what the moral of the story was, but I really liked the lore, aesthetics, and world building. I'm so glad someone is bringing my favourite sci-fi to life! We really do live in the future!
Wait I can’t tell if this is genuine or not
@@Penguinman2.0Neither can I…
On a second reading, I don’t think it’s genuine, “I don’t remember what the moral of the story was” seems like it wouldn’t be there if it was.
@@Penguinman2.0 The book doesn't exist, its from a meme talking about how techbros proudly claim inspiration from scifi stories where the whole moral is NOT to build the technology featured in the movie. Like how vr/metaverse nerds constantly say "its like the Matrix in real life"
@@SissypheanCatboy or more recently, openai comparing gpt-4o to the movie "her"
@@SissypheanCatboyThe first thing we did after GPT-1 showed promise was hook up the next version to the internet to learn from. It's like the people at the forefront of advancement have turned dystopian prophecies into a checklist..
If youtube was just this type of content but with every expertise/experiment currently going on. It would be just perfect harmony.
The perfect combination of long-form content, diligent and passionate work, and humor!
Meanwhile RUclips: Demonetized.
TestTube
@@yaysuu holy smokes you're onto something!
not only that, it would inspire young people from age 2 onwards to become youtubers and make the next big scientific discovery/invention!
Guys, don't stop. I NEED to see what will come out. Your first 2 vids inspired me so that from just a noob in programming thinking to himself "well, im gonna be a programmer and program commercial things" you made it so I now have a greater purpose in life and right now I'm doing an EEG research together with a neurobiology lab (I was the one who contacted with them and suggested a research and I'm the one doing it, but they give me access to some fancy things). Thank you and DON'T STOP UNTIL ITS DONE
>wake up
>your entire existence is DOOM
>pass on thinking that was all there ever was to life
Same as it ever was.
>wake up
>your existence is this one
>pass on thinking that was all there was to life
@@poupeuu This is why I choose to participate in 5 dimensions
@@jeremymcadam7400 rookie numbers, all of us in the 6th dimension are looking down on your struggles, purely for amusement I might add
Plato’s “The Cave” is Doom.
"Yeah I'm doing research on neural networks."
"Oh, what algorithms are you working with?"
"Rat"
Underrated 😂😂
Can it run doom?
That's the medium, not the algorithm.
Rotational
Additional
Tactics
RAT
🐀🐀🐀
🐀. 🐀. 🐀. 🐀🐀🐀🐀🐀
🐀🐀🐀. 🐀🐀. 🐀
🐀. 🐀. 🐀🐀🐀. 🐀
🐀. 🐀. 🐀. 🐀. 🐀
and where do you work?
A weapons corporation..
9:29 "Why those ingredients? I have NO IDEA!"
Yep! That's how you know someone is doing real science and not just teaching you prepared lesson.
(*Cave Johnson enters the chat*)
@@johnpooky84PORTAL MENTION
Hey! I work with mammalian cell cultures, and I know they can be very finicky! If you didn't change the media a day after your initial seeding, the residual DMSO from the cryopreservation media is likely what killed your cells. Not sure if you did this or not, but I didn't see it in the video so figured it was worth mentioning! You guys are awesome, keep up the great work.
24:40
Surely this is the worst O-Ring disaster in the scientific profession...
New challenger appeared!
@@noi0124it’s the challenger space shuttle ! Ready? Set Fight!
Maybe they just have to chill… *checking notes*
On no, wait. That was NOT the cure 😆
Too soon
Context? (Curious)
My other suggestions seem to have been deleted, perhaps because I linked a paper of ours. I guessed that you may be killing cells with too much light. Phototoxicity is a huge problem with any fluorescence microscopy. To reduce it, use phase contrast (not fluorescence) microscopy and use longer-wavelength (red) light. Also, when doing fluorescence imaging, use a lower-NA lens, more sensitive cameras, dimmer illumination, longer wavelength excitation, and shorter exposure to light.
Feel free to send me an email with all of the suggestions so they don't get lost! And they're greatly appreciated as always.
We only use phase contrast when checking on the cells. Fluorescence was only used at the end for the sake of the experiment, and the dye was only used on one of the arrays after we had finished all other experiments. The arrays are also only ever left on the microscope for as brief a moment as possible, but the phase contrast images aren't as pretty so I don't use that footage in the video as much.
I also saw you recommend concentrating the neurons in the middle. I read that in the instructions and that's how we attempted it the first time when the cells died. We went with adding the neurons after the media because we knew for sure the neurons would survive as we've tested that method a lot and couldn't risk another culture dying. We were pressed for time to get the video out. And for our arrays the electrodes are REALLY spread out, so we're gonna need a high neuron density across most of the surface of the array anyway. You'll notice I made the walls of our arrays taller than the multichannel ones. It was specifically because I wanted to increase the amount of growth media per well so the neurons don't deplete it as quickly and to account for an eventual much higher neuron density across the array.
We have wanted to upgrade our camera on the microscope for a while, but the scope itself was a HUGE upgrade. It's new for this video and it cost a pretty penny. But it's built exactly for this kind of experiment so we can switch out the camera next. I also want to move to a red dye to lower the illumination wavelength as you say. And also just to make the videos look more interesting than always green. We ran out of time, but we also got a tubulin specific dye we're gonna show off next time that would look best against an otherwise red stained neuron. And we picked up NucBlue so we can stain the nucleus properly. One of the shots of the neurons firing included it from the first well we tested it on. I overlaid it. It's subtle but makes the neurons look nicer and the nucleus visible.
Also, I've set your account to always show comments on the channel now, so they shouldn't get deleted.
@@thethoughtemporium youtube doesn't care and if the automated system even imagines your comment as bad it will just automatically delete it or hide it
10:09 "Hey how are the neurons growing?"
"They all died"
*AD*
...had a hard time trying not to chuckle in the doctor's office.
Hope the appointment went well! God bless!
What
Neuron: Help I'm peeling
Neurobiologist: SUGAR FREE HORSE JUICE
Just want you to know that your channel is what introduced me to the field of biological engineering. I'm now pursuing a doctorate in the field. Thanks for being an inspiration.
DOCTOR STEVE POTTER CONTACTED YOU!
THE DOCTOR POTTER!
This is surely indicative of success to come
For the ones randomly watching this and has no idea who Steve Potter is and why it is a big deal
It is as if Jesus Christ himself showed up for a theology class
A truly impressive feat!
What a Legend.
I’m sure he’s never heard this joke, but he sounds like a wizard in this field.
@@akatsukilevi I can't not stop thinking about Professor brothers now lol
Neuron-piloted flesh mechs growing neurons, simulating neurons, looking at neurons, visualizing neurons.
Ah yes, Neuron Activation.
Soon we shall have a flesh automaton animated by neurotransmitters!
Do not dare to insult the purity of steel by comparing mechs to abominable flesh
Bro just called God's greatest creation "neuron-piloted flesh mechs"
@@Mate_Antal_Zoltan It's a Cruelty Squad reference xP
@@audriusskebas3099 guy who has only played Cruelty Squad in his entire life: "I'm getting real Cruelty Squad vibes from this..."
I had no idea you could actually grow live neurons and train them to do stuff... thank you for documenting your unique experiment and sharing it with the world.
This part of RUclips is interesting because you have genuine, field respected scientists contributing to a meme. A similar situation I see is a video series an amateur arachnid enthusiast making absurdly detailed videos about common house spiders, and basically has tons of scientists conversing with him and in the comments about the most trivial of differences between two species of harmless spiders
Which channel is this?
@@Astoria_Varanus Oops, I forgot to put his name: Travis McEnery. The series is called "The Spiders in Your House."
To be fair, if this project succeeds it will be the ultimate, unbeatable iteration of the running doom on X meme.
13:41 It does say "Minimize light exposure" on the bag.
True. 😄
Hey! I’m messing with neurons at the moment in my PhD and I saw the exact same morphology in DMEM/F12, I was stumped what was causing it! As a precaution we’re genotyping with ICC and qPCR since we’re deriving from iPSCs - anyway, love the video! This project is so cool!
dr stone would probably accomplish this with the power of jump cuts and eye closeups
Power of anime
And that's why he is called Dr Stoned.
I loved that show! Hoping for a Season 3.
@@johnpooky84 I was wondering the same thing and I think there is a season 3 but it’s not on netflix yet?
@@Numbers-gStands Dr. Stone's on Netflix? I watched it on Crunchyroll.
I am a mitochondrial and cancer physiologist, and not a neurobiologist, but have been studying metabolic and growth effects of different commercial medias. It might be worth keeping in mind that the metabolite concentrations in DMEM are largely not found in the brain. I'd be curious to know how Neurobasal and DMEM compare in their composition. All of these considerations, i.e., metabolic activity and ionic concentration, will affect the neuron's membrane potential.
I WAS THINKING THE SAME THING!
DMEM with FBS (foetal bovine serum) is an excellent medium for accelerated growth but yeah, it's 4x stronger than standard EMEM. EMEM or Opti-MEM may be a better option for maintaining mature cultures, with Opti having the added benefit of requiring much less FBS.
You are one of the main reasons I am perusing my masters in synthetic biology!!!! I love your videos man hope you keep doing what you do :)
The trick to finding a comfy spot for your tongue is actually the roof of your mouth. People struggle with it because it's not intuitive that you keep it up there with suction normally.
μ
i love mewing
Huh, it’s kind of comfy
Funny enough I have a mint in my mouth and was already moving my tongue
Thanks!
"What if we grew a mind in a vat that only knew how to interact with the world through violence and was trained to snap headshot imps in .03 seconds, this will surely not go terribly wrong."
Yeah, this could never go wrong, but to make sure lets wire a 100 of these up to the Boston dynamics robot wielding a shotgun. But we would still be in control, right? lol
I know this is a joke, but the interaction that the neurons see is not in any way related to violence. it's like if you played doom, but instead of a screen, you just see an array of 10 blinking lights and no-one told you that you're playing doom, only if you're succeeding at pressing the correct buttons or not.
so ... as long as no-one tries to apply this to real-life weapons, there's no reason whatsoever to worry. though ... now that I think of it, knowing humans, this does indeed have me worried
@@DrippyPootis
Boston dynamics V1
Context- For it to work like the joke intended it would need some reason for violence like a reward system but for that to work we have to have some sort of detection software supporting it and at that point just use software, so yeah the joke falls apart, oh well. Except for machine learning being applicable to organisms, i.e. stimuli training, so maybe we could consider culturing them with positive stimuli for shooting the target before sending them but that sounds like a lot of work. Not that someone wouldn't try? Also if the logic reply wasn't to the Boston Dynamics bot joke then that is really embarrassing lol
@@DrippyPootis on the other hand, the neurons not being aware of it being violence points to another issue, since that means they won't have any concerns whatsoever with performing violence if put in a situation where they might accidentally do so.
putting any kind of neural network, artificial or biological, in charge of dangerous equipment means that unless you very actively train it to avoid harming humans, it will do so if any humans get in the way. this is of course already the case with any kind of automated machines, even without neural networks or similar, which is why we tend to place robots in cages, so we can better protect humans from them
I think we are not appreciating the work they are doing: THEY ARE LITERALLY BUILDING A CPU FROM SCRATCH. Something that costs companies with billions of dollars and hundreds of engineers, but on top of that they are doing it with neurons. I mean MY HEAD EXPLODES THAT THEY DO THIS AND SHARE IT. Seriously, thank you very much, I have watched the videos in this series dozens of times and I am obsessed with this technology, you inspire me. THANK YOU!
*Rip and tear playing in the distance* THE TIME HAS COME
Rip And Tear? So, R. A. T
if youre still having issues with the array electrodes being cut correctly, maybe ask a glitter company if they can help. I know its a weird idea but glitterX is just a company with precision cutting instruments. Might be worth it
Wasn't there the old riddle about what the glitter companys biggest customer was and they have absolute secrecy and nobody knows? 🤔
@@harriehausenman8623 Yes, and it is most likely pearlescent paint for hobby boats. Boat manufacturers don't want it to be associated with how "unmanly" glitter is seen as, as well as the fact that pretty much all of it ends up as pollution in water.
Impressive! It’s really nice to see the path your channel has taken. From the time I was watching you hunting VHF satellites in Brazil, Wi-Fi imaging, the pulsar corner reflector (that one got me really excited), to DNA hacking and neuron training. I would like to say I came for the RF and stayed for the bio. 10/10
Thank you Multichannel Systems (affiliate of Harvard Bioscience)! Without your help we never could have built the Torment Nexus.
Hey... I've noticed sometimes when pipetting, you (or whoever is doing the pipetting) touches the destination, and they go back to the source. This is a no-no as it can cross contaminate the whole batch, and the source material. I'm not even talking about living contamination. In the examples I saw, I don't think it's that big of a deal, it would introduce neurobasal into the neuron source.
But it's a bad practice either way.
Yep, once it touches a new source, tips go bye bye. I'm surprised at some of the stuff done here, as we ALWAYS filter our media... And I haven't seen that. Also, just pouring tubes out can be a source of contamination.
This is one of the most impressive RUclips project I’ve seen
Once again, casually bringing cutting edge research to RUclips. Legend
THE DAY HAS COME BROTHERS! 🗣️🔥🔥
FINNNNALLLLLYYYYY
LETS GOOOOOOO💯💯
Ikr! I've been waiting for this for what seems like FOREVER!!!!!
yay
WE WILL FINALLY MAKE A MEAT COMPUTER
I didn’t know that there a so many materials in common with 3d printing! Fep, pei, teflon, lactate. Just waow!
18:13 putting a tinfoil hat on the neurons is amazing
Can't wait until 2030 when I have to worry about giving my phone a concussion when I drop it
I've been waiting almost a year for this video! Now it's finally here! Thank you so much! 🎉🎉🎉
Love the Tumblr post reference with the Torment Nexus joke
This is easily my favorite Science channel. Keep up the work, this is amazing content
This is probably the coolest experiment I have ever seen - I am really looking forward to the next part!
Another interesting aspect you could add to this would be to have two separate cultures of neurons acting on the same system. This would allow for demonstrating coordination and collaboration between independent cultures for instance.
you have no idea how excited i am about this update. I've been rewatching the old video very often as a source of comfort
neurons: WHY IS IT SPICY
NOT THE SPICEEEE AAAAA
Teşekkürler.
18:13 don't forget to wrap the wires too, it's how you turn a moderate speaker system in to a high end speaker system.
The wires look like they might be fiber optic?
Apart from USB, but that’s error corrected
the most human thing ever is to build horrors beyond their comprehension just to spite some book
you know, when it's all laid out like this, it makes it sound simple
One of the few time I actually waited for a video to come out and the concept is cool af, hell yeah!
Today is a day to remember. Tomorrow is a day that we will never forget
I can't remember today. It is currently happening. I can't remember tomorrow. It hasn't happened yet. Soon I will forget about yesterday.
@@MaxBrix bro forgor 💀
14:28 ah yes, 2037 exam cramming by putting chilli flakes on the brain. Effective
ON TODAYS EPISODE OF MANMADE HORRORS WITHIN COMPRENTION
(edit: i will respect english spelling once its concistant (was that wrong, probably) fake ass language)
ikr
How many todays are there?
There’s no need to shout.
COMPRENTION
spelling is beyond your own comprehension
I totally agree with u tbh
Do not let bro start his villain arc 💀
Too late buddy 😂
Hey what was the unedited?
Adding capsaicin to the culture was the start of the villain arc
Wow, this comment is hilarious and I really appreciate the originality😐
@@Bread78787 shh-
For your cap problem. Get a smaller radius plug to fit in the collection. Use the semipermiable membrane as the gasket layer. You can then place an o ring directly on the plates acrylic pipe.
Damn, that poster looks AMAZING!
Thank you!
This is quickly becoming my favourite series on RUclips
“0:00 this is your brain. 1:22 this is your brain on [Petri dish]. Say no to [chemistry]”
Cannot wait for growing brains to beat Super Mario 64
Donkey Kong is next!
Now THAT would be something to see.
my friend sent me this and i a m GENUINELY not at all upset, i love this shit i am going to binge ALL OF IT
What if we somehow connected neurons to a limb to train it to hold and shoot a gun, so that the neurons could kill a human being.
Bro said my intrusive thoughts lmao
Robobrain is the future.
yoo yo have the same name as my classmate
@@oightKoreraAreEditable Really?
arrest the neurons
"This is your brain...and this is your brain on doom. Get the picture?"
In the words of 4 year old me, "I like the spicy chemical."
2:20 That tech tree reminds me a lot of Dr. Stone. I realise it's also a common enough trope that it could have come from anywhere, but the specific style of this one looks distinctly inspired by Dr Stone's approach to the idea.
Wonder if the neurons will freak out when they cant look up or down.
only as much as you'll freak out when you realize there's a fourth dimension you can't rotate yourself in
@@rootabeta9015 Did somebody say 4D Golf? 😄
@@rootabeta9015actually I’m rotated in time by 4°
@@rootabeta9015 sounds like a skill issue, I can rotate in the 4th dim just fine
@@harriehausenman8623 non-intuitive double rotation intensifies*
I was about to cut a chunk of my brain to experiment by myself BUT PART 2 CAME TO LIFE SO NOW I CAN LIVE WITH MY FULL BRAIN!!!
DO IT ANYWAY!
They write a book about why you shouldn't make the torment nexus and you STILL make it 😭, that's nefariously doom-like
2:18 "don't build the tormentor nexus. And thus far we've been making great progress"... wait wait, so, you are making great progress in NOT BUILDING the tormentor nexus, or in building it?
Yes!
@@thethoughtemporiumwhich one?
And we're back on the super villain type beat
This is one of the coolest projects I've ever seen. Who knows, maybe someday you'll connect whole rat brains to the internet
It'll end ul learning to Google for pictures of cheese and female rats.
@@carlpanzram7081 and cat slander.
Ah yes finaly update for the literal game of life😂
the devs take a while to update the game
Horton Conway would approve this comment. 🙏 R.I.P.
"I want to build up to simulating a whole brain" followed by "...so anyway" caused me at least some level of concern.
i had no idea this was out, ive been waiting for this for ages
From now on I shall only respond to "slightly sentient enchilada"
Admittedly I only understand SOME of the concepts you talk about (way above the basic bio from my degree), but man this is sick as hell, I love it
It's sick, and.. you love it?...
@@cnnhean sick is slang for cool/good/rad in some places ☺
@@afinnishfishnet7366 you maybe right in this context.
For me messed up is the accurate representation of growing neurons for such purposes.
@@cnnheanthe neurons can't feel. It's hundred times less messed up than how most of our animal products are produced.
I want to remind you all that this chad literally genetically modified his own DNA purely to consume pizza again.
“What’s my purpose?”
“To play doom”
“Oh my god”
This is a certified "What is my purpose?" moment.
*to play doom.*
18:09 “slightly sentient enchilada” made me inhale half my beer
It came ...... after 3 years
It did!
I love that they are doing a entire scientific experiment to play doom it is beautiful
It's funny because a difference between a video game and a live camera feed are the same to this. I bet someone is already leaps ahead doing what you are doing but you are absolutely on the right track.
Imagine having a CPU that's literally just a brain, how fast would it compute compared to a normal cpu?
And hopefully it won’t gain senti ence
Imagine it grows new connection base on the things you do on it, gradually speeding up.