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[Music]

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hi everyone so my name is Tony Jaa I'm a

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researcher at the earth Life Science

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Institute at Tokyo Institute of

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Technology LC for short and I'm also an

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affiliated researcher with Blue Marble

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space Institute of science I'd like to

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thank Mike for such a great musical

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introduction and then I'd like to thank

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the organizers for giving me a chance to

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share my research thank all the audience

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for staying awake for this in the next

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talk there's two more great packs so

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keep it up and then I'd really like to

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thank all of my abundant YouTube and

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Facebook fans following online if there

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are any anyway so yeah so today I'm

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gonna be telling you a little bit about

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some research I've been doing at LC

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regarding polyester micro droplets and

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their role as compartments similar to

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AMA this actually was published

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yesterday so please read this online

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also afterwards if you have any interest

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so we make a very big generalization

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here so biology is very complicated and

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biology is made of cells and cells are

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very complicated cells have a ton of

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different functions and very specific

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organelles which have been tailored to

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perform very specific functions so many

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modern biologists and biochemists are

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really interested to study how exactly

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the cell functions and how exactly it

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formed I'm gonna again make a very gross

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oversimplification here and assume this

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is a modern cell all right it's not this

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simple but basically you have DNA

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genetic material that turns into RNA and

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RNA makes proteins which are you have

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enzymes you have very functional

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catalytic molecules and these are all

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enclosed within a lipid bilayer this is

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like the compartment that encloses all

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of these things what did an early cell

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look like so well we don't really know

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exactly

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actually how modern proteins could have

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evolved in assembled we don't really

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understand how DNA came about and so

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many people do believe that primitive

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cells are some type of lipid vesicles

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where with RNA inside I'm gonna skip the

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next part because Krishna did a really

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good job to explain the RNA world

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hypothesis and basically what what we we

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understand is because RNA was genetic

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information and could perform catalytic

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functions it's been proposed to be one

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of the first early biomolecules on the

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early Earth now furthermore though one

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one question that many researchers have

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had is you know how exactly did RNA

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replicate there's been a lot of research

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progress no in this field we still don't

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have a really concrete answer and we

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don't necessarily have a very concrete

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answer about how this lipid vesicle type

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bilayer vesicle form we don't even

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really have a concrete answer of how

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exactly RNA could have formed so you

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know what on earth happened whether you

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you know understand the RNA world to be

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correct or not we still need to

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understand you know even if it were

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correct what about before or such a

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world existed what happened on earth

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before vesicles what happened before RNA

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and so these are really interesting

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questions that I've been very I've been

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working towards at LC one of the

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important top things that you should

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understand is there are many ways by

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which biomolecules could have formed and

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assembled on earth that we as we've seen

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with many of these talks they could have

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come from extraterrestrial sources could

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have been synthesized from spark

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discharge or through catalysis by

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mineral surfaces and so there's a many

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ways by which you know nucleotides or

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amino acids or lipids could have could

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have been formed on the early Earth and

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one of the things that's very important

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to understand is that when you're

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forming these biomolecules you're also

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at the same time forming many non

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biomolecules there's many bio mode many

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molecules forming at the same time

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as these relevant biomolecules through

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all of these processes and so what I

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wanted to know was can non biomolecules

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catalyze life's immigrants in some ways

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can we take all of these you know extra

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things that also existed on early Earth

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and can that assemble into something

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interesting and important for the

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emergence of initial life I think Amy in

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her talk talked spoke briefly about

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alpha hydroxy acids basically

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structurally they're very similar to

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amino acids their differences one

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functional group and what's interesting

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about hydroxy acids is even though they

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don't participate in if you understand

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modern biology to be DNA are native

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proteins they don't participate in that

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type of process that which is so called

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central dogma but they are very

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pre-medical abundant and they also are

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compatible with modern biology so you

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can actually replace amino acids in the

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translation system that forms proteins

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and you can replace them with alpha

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hydroxy acids and what you get is in the

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end polymers of alpha hydroxy acids

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called polyesters so we know that these

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alpha hydroxy acids these monomers are

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very common prebiotic aliy and they're

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compatible with modern life and so what

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we wanted to do was we wanted to take

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many different types of alpha hydroxy

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acids I don't worry about the detail so

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much but understand that we took many

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different chemical functionalities and

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we took many different alpha hydroxy

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acids with different chemical

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functionalities and in a complex mixture

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we wanted to utilize wet drying cycles

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to form polymers so monomers are one

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circle polymers are many circles kind of

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shove together and we wanted to form

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polymers and see if these polymers could

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have assembled into something useful for

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life's emergence so these are the five

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that we tested and we first took

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mixtures some

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missions of each of these they're

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concentrated solutions and water and we

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dried them down

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drying is reasonable primitive process

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we dried them down and we wanted to

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analyze the product so we did that by

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taking the products and subjecting that

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to something called MALDI ms so matrix

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assisted laser desorption ionic mass

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spectrometry basically we took a solid

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and we analyzed the components of that

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chemically through measuring the masses

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of whatever was in there and you read

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this by the x-axis is the mass of a

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given molecule and the y-axis is the

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abundance and what you end up seeing is

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that for example here this peak

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represents one molecule which is mass

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you know like 560 this peak represents a

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molecule of mass 600 something and what

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we saw was that after this

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polymerization process we didn't just

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make one product we made a variety of

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products as polydisperse products they

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ranged in size from you know a few units

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long

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up to the 40's and each peak was the

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difference between each people was the

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correct mass for each monomer unit and

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so that's how we knew that each one

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corresponding to a specific polymer

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links so yeah we were able to create up

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to you know in this case for lactic acid

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up to 44 units long polyesters and we

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did this for all five of these

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combinations and in each case you do you

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produce not just one compound you

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produce a poly dispersed mixture of

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compounds and the maximum length of each

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were differed between 15 and 40

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depending on each of the monomers that

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you used what we did see actually was

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after drying this is about after a week

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we saw we found this you know gel-like

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substance and we saw this for almost all

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of these samples we tested so for four

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out of the five homo polymer samples

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that is the samples with one

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of alpha-hydroxy acid in the starting

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mixture these all formed the gel-like

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substance and also when we mixed all

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five of them together and subjected it

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to the drying process they also form the

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gel like structure in one case poly

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glycolic so Polly glycolic acid did not

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form this gel like structure there's a

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mechanistic study that I won't get into

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right now so then if you formed this gel

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structure so on right so what the next

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thing we did was we actually highly

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hydrated these gel like structures it's

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been shown that with this alpha hydroxy

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acid polyester system you can do

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repeated rehydration and drying cycles

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and you actually get different sequences

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each time that assumes that the sample

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isn't dried to completion in this case

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it was actually dried to completion we

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believe which formed this Joe X

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structure and upon rehydration

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if assembled into these micro droplets

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each homo polyester formed micro

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droplets except the one that didn't form

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of the gel and the mixture also formed

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the micro droplets interestingly all

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mixtures of 2 3 4 or 5 all combinations

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of those all formed micro droplets and

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we believe that you can probably add in

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some other chemical functionalities and

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still form similar types of droplets and

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so what you what you should get from

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this is just that we can form polyester

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gels from drying of alpha hydroxy acids

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and they can assemble into micro

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droplets in aqueous media we're also

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interested to see some of the dynamics

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of these droplets so one of the

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important things that were in we think

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is important for early life is the

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preservation of individuality how long

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that can separate droplets stay before

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they merge together and so this is just

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a video at their ambient conditions and

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you can see that maybe it's a bit grainy

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but the droplets don't easily don't

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quickly call us

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on the order of minutes hours it takes

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about a day or so for them to call us

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and to kind of a large microscopic

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droplet but when we added salt we

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actually saw that the droplet started to

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coalesce much more quickly and they

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actually stuck to the glass surface in

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the microscope we believe that this is

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because the salt and some researchers

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have done simulations on this that we

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believe that the salt is locating to the

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outside of the droplets it's lowering

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the surface tension of the droplets and

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that's causing them to kind of merge

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together a bit more and stick to the

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glass surface quickly and further

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studies are ongoing into determining

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exactly this mechanism including

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measuring directly where these salts

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ended up and then there are some also

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implications of these droplets as

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compartments as you know cells contain

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many things in the modern cells genetic

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material enzymes nutrients primitive

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cells protocells also necessarily had to

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contain some type of catalytic or

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informational polymer in them and so

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these droplets we first we want to test

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whether they could encapsulate things

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and we the easy thing to do was we found

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some fluorescent dyes and we saw that

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these fluorescent dyes were able to

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segregate into the droplets the dyes are

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fairly hydrophobic and we believe that

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the interior of the droplets is also

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very hydrophobic owing to the very long

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polyesters that produce them but also

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we're able to see that fluorescently

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labeled RNA is able to go into one of

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the droplet types which suggest that

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perhaps one of these droplets or some

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droplet type that we haven't probed in

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this study could have been able to

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compartmentalize and segregate primitive

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genetic materials and so this also has

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implications that the initial chemical

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components of these alpha hydroxy acids

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that we use to make the droplets will

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determine their

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phenotypic function if you will because

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only one of them could segregate these

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fluorescent RNA and just a few more

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examples of the interaction between the

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droplets and biomolecules we took some

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super folded GFP that was expressed by a

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collaborator at LC he's in Sean McGinnes

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lab that name has been floating around a

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few times and he provided us some super

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folded GFP this actually function in the

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droplet suggesting that even within this

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confined space the function and the

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structure of these this protein could

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still be preserved and we were able to

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see that certain fluorescent lipid could

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assemble layer around one of the

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droplets we're not sure whether it's a

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single layer by layer multi layer maybe

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it's a multi layer but we can see that

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this fluorescent lipid does have some

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propensity to assemble around this

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droplet suggesting that maybe the

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droplets we whether or not they were

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direct precursors to modern cells or

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primitive cells or not and possibly they

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weren't they could have still catalyzed

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the assembly of some structures that

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could have been providing important for

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the initial emergence of life and so

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with that said I'd like to just go over

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this really quickly we started with a

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monomer solution we dried by heat to

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form polymers polyesters upon

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rehydration they form micro droplets and

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these micro droplets are able to

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compartmentalize certain dies and

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biomolecules and so what I hope that you

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got from this study is that non vesicle

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compartments are also important for

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origin of life studies and that even

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non-biological molecules could have had

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a hand in producing important biological

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structures or materials or molecules

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that could have helped in the initial

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emergence of life so I'd like to thank

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all my funding Oh my collaborator

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especially those in both

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who directly contributed to this study

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and I'm from Elsi were an

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interdisciplinary Institute in Tokyo

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that's interested in studying the origin

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and evolution of planets earth and life

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on earth and so if you have any

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opportunity please I hope you can visit

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we have an annual symposium every year

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next year's will be in February so I

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hope that you can if you're able to

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please join us so thank you very much

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for your time and attention and have you

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00:16:07,400 --> 00:16:05,620
take any questions questions for Tony so

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00:16:12,900 --> 00:16:07,410
what why do you think you've got that

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Gaussian distribution it was like Bill

355
00:16:26,400 --> 00:16:21,339
like okay so you're referring to then

356
00:16:30,960 --> 00:16:26,410
you got yeah okay this one yeah so if

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00:16:35,210 --> 00:16:30,970
you assume that the condensation of an

358
00:16:38,639 --> 00:16:35,220
additional monomer piece is somewhat

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00:16:41,400 --> 00:16:38,649
stochastic then perhaps you get some

360
00:16:45,900 --> 00:16:41,410
type of distribution where there is like

361
00:16:47,910 --> 00:16:45,910
a medium amine and there fewer molecules

362
00:16:50,430 --> 00:16:47,920
that are larger and fewer molecules that

363
00:16:53,939 --> 00:16:50,440
are smaller and this is also helped by

364
00:16:56,370 --> 00:16:53,949
the fact that this conjugation reaction

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00:16:59,340 --> 00:16:56,380
is not one direction it's bi-directional

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00:17:01,380 --> 00:16:59,350
and so you do get once you form the

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00:17:05,240 --> 00:17:01,390
polyesters once form the polymers you do

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have some backwards reaction as well

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00:17:14,039 --> 00:17:09,040
thank you Tony I had a couple questions

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00:17:16,530 --> 00:17:14,049
um so this was just one drying it wasn't

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wet dry cycling is that correct yeah can

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00:17:19,380 --> 00:17:17,949
you speak about what you think would

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happen if you were cycling a little bit

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00:17:26,390 --> 00:17:22,209
yeah so I think the cycling question is

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00:17:29,880 --> 00:17:26,400
heavily determined by whether the

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initial reactants have completely dried

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or not if you're still in a mostly

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00:17:37,430 --> 00:17:35,430
liquid state then by rehydrating your

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00:17:40,970 --> 00:17:37,440
shifting the equilibrium of the reaction

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from formation of polymers back into

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00:17:44,240 --> 00:17:42,360
monomers and this is because the

382
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formation of the polymers requires loss

383
00:17:51,020 --> 00:17:48,630
of water but if you're in this gel soft

384
00:17:53,120 --> 00:17:51,030
state kind of not quite liquid not quite

385
00:17:56,320 --> 00:17:53,130
solid state maybe there is some

386
00:17:58,880 --> 00:17:56,330
hydrolysis and I think it depends on the

387
00:18:01,790 --> 00:17:58,890
chemistry that you've used specifically

388
00:18:04,760 --> 00:18:01,800
but also perhaps the fact that you're

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00:18:07,700 --> 00:18:04,770
forming these kind of aggregated

390
00:18:11,270 --> 00:18:07,710
structures the surface area that water

391
00:18:13,430 --> 00:18:11,280
can attack if you will is may be limited

392
00:18:14,960 --> 00:18:13,440
by formation of these structures so we

393
00:18:16,420 --> 00:18:14,970
haven't done any of these studies yet

394
00:18:21,140 --> 00:18:16,430
but we're interested to study whether

395
00:18:23,690 --> 00:18:21,150
actually assembly can prevent their own

396
00:18:25,490 --> 00:18:23,700
degradation and also if they can prevent

397
00:18:27,490 --> 00:18:25,500
the degradation of things that have been

398
00:18:31,100 --> 00:18:27,500
compartmentalized within the droplets

399
00:18:33,320 --> 00:18:31,110
and then I also just wanted maybe I

400
00:18:34,640 --> 00:18:33,330
missed it but what was the scale bar for

401
00:18:37,610 --> 00:18:34,650
those images what are the sizes of all

402
00:18:40,940 --> 00:18:37,620
yeah good question sorry I actually

403
00:18:46,760 --> 00:18:40,950
completely skipped that so these images

404
00:18:48,920 --> 00:18:46,770
are all so the main image scale bars so

405
00:18:52,190 --> 00:18:48,930
those down here this is going to be a

406
00:18:54,260 --> 00:18:52,200
hundred microns and this is ten microns

407
00:18:56,960 --> 00:18:54,270
so they range in size from a few microns

408
00:19:00,050 --> 00:18:56,970
up to tens of microns you know 50 or 70

409
00:19:02,390 --> 00:19:00,060
and it does I think it depends on the

410
00:19:05,330 --> 00:19:02,400
chemistry involved but also you can make

411
00:19:25,520 --> 00:19:05,340
them smaller by sonication and vortexing

412
00:19:28,370 --> 00:19:25,530
and yeah so we were gonna be starting to

413
00:19:31,250 --> 00:19:28,380
do some of these this work actually so I

414
00:19:32,630 --> 00:19:31,260
mentioned that we could assemble a lipid

415
00:19:34,520 --> 00:19:32,640
layer that was a phospholipid layer

416
00:19:36,800 --> 00:19:34,530
around perhaps assembling this type of

417
00:19:38,690 --> 00:19:36,810
layer mono layer by layer multi layer we

418
00:19:40,280 --> 00:19:38,700
don't know but perhaps assembling this

419
00:19:44,300 --> 00:19:40,290
type of lipid layer around it could

420
00:19:45,270 --> 00:19:44,310
prevent a coalescence and also maybe

421
00:19:47,250 --> 00:19:45,280
prevent

422
00:19:50,130 --> 00:19:47,260
you know the exchange of things in and

423
00:19:52,049 --> 00:19:50,140
out of the droplets but also there are

424
00:19:54,840 --> 00:19:52,059
things called Pickering emotions which

425
00:19:56,460 --> 00:19:54,850
are basically if you have some type of

426
00:19:57,960 --> 00:19:56,470
phase separated system there's these

427
00:19:59,490 --> 00:19:57,970
little particles that locate

428
00:20:02,100 --> 00:19:59,500
specifically to the outside of the

429
00:20:03,960 --> 00:20:02,110
droplets that prevent coalescence one of

430
00:20:05,669 --> 00:20:03,970
which is it's an interesting study by

431
00:20:08,549 --> 00:20:05,679
Christine Keating's group at Penn State

432
00:20:10,350 --> 00:20:08,559
where they use clay particles around a

433
00:20:12,120 --> 00:20:10,360
quiz two-phase system droplets to

434
00:20:14,310 --> 00:20:12,130
prevent their coalescence so there are

435
00:20:17,370 --> 00:20:14,320
some like chemical or physical ways

436
00:20:21,870 --> 00:20:17,380
perhaps that these could maintain their

437
00:20:25,289 --> 00:20:21,880
individuality any other questions for