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[Music]

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hi everyone

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um I'm just going to make a quick note

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before we start that it's great to be an

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astrobiology conference because the

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background slides are going to go much

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more smoothly I presented the sort of

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microscopy conference a few months back

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and it was a different experience so

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thank you all for being here

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um and being you

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um so

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um I'm going to talk uh this is going to

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be a little bit of a pivot from some of

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the previous talks but I'm going to talk

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um about some of the methods that I've

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been working on to actually detect life

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on a mission a life detection mission to

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Europa or Enceladus I'm also going to

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make a note so I'm going to be talking

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about uh a life life detection missions

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to Europa and Enceladus as a target for

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this but um once you have a space

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capable microscope this could

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potentially be applied to like all sorts

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of destinations so there's lots of fun

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possibilities here

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the aforementioned background slides so

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um when we're picking destinations to

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look for life

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um

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there's uh we like to look for places

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that have habitable conditions things

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that have liquid water abundant energy

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sources and conditions that are suitable

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for uh Life as we know it or as we might

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predict it may exist and

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um I'm just gonna let Enceladus and

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Europa sit up there because we've

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already heard a lot of justifications as

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to why those are promising targets to

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find those conditions and why we might

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have had

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um an uh origin of life or uh living

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organisms currently there

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um but when we're searching for life we

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also have to think about how we're

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actually going to look for it and so

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um we can kind of break down life into

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some of the key components that make it

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up things like uh catalysts that help it

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uh metabolize

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things like information storage polymers

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so DNA or RNA and earth-based life and

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that might look very different for

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extraterrestrial life

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um and also compartments life on on

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Earth Life as we know it loves to put

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things inside of membranes and then

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sometimes they put those membranes

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inside other membranes and have them

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connect to other even more membranes but

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those that packaging really helps it do

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some interesting chemistry

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um and so when we're then thinking about

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this in the context of a mission

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um we can start to think about how we

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might actually observe that and what

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analytical approaches we need to take to

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try and get at that

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um and so I put the like giant table up

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here

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um because uh these are all things that

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people are working on

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um for Life detection instrumentation

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but we're going to be focusing for this

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talk on containers

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um and so

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um the reason for that is

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um that again as mentioned life loves to

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package things into compartments and

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those compartments can be directly

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observable by using a microscope to look

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for morphology and also for the

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collocalization of certain key chemical

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biomarkers

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um

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so in

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um our team we're working on um a

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microscopy system that we call a

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luminescence imager for exploration or

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life

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um and there is uh two key hardware

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components of this system one is the

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fluidic subsystem and this is to deal

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with

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um the actual sample processing and

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preparation for Imaging something so um

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my background full disclosure is

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actually not an astrobiology it's in

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fluorescence microscopy and

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um you know I spent like five or six

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years in grad school just moving fluids

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around in tubes and like preparing

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things on slides before they actually

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get to the microscope I've been told it

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would be expensive to send me to

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Enceladus so uh this is this is my

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replacement

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um and then the other end is the actual

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microscope itself so the optic system

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which is going to have all of the things

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that you need to actually take your

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pretty pictures

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um so I'm gonna break down what goes

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into these systems a little bit so the

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fluidic subsystem that uh like uh silver

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plate on top

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um is the rotary filter stage and so the

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um the stage can do seven individual

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experiments um and so that accounts for

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originally this was designed with the

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Europa Lander Mission Concept in mind

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where there would be three separate

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samples each with a replicate and then a

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procedural blank and then each of those

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individually has

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um

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three uh filters of different sizes

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um and so uh that basically will let you

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sort out particles so the filters are 10

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microns 1.2 microns and uh 0.2 microns

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in size and that's just so you don't end

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up with larger particles obscuring any

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of the smaller particles that you're

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trying to image in your sample

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um and that way you're not missing any

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of the like good stuff that might be

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there

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um

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there you use all it's also attached to

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the fluidix manifold which is the part

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that actually moves all of the liquids

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and fluids around and it also has a lot

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of sensors for pressure pH conductivity

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and also crucially some storage for the

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fluorescent stains that I'm going to be

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talking about a little bit later in the

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talk

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um

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and then these were the filters that I

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mentioned earlier and so uh this is

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again for sample size sorting but

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they're also made of a silicon nitrate

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substrate that has a low fluorescence

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background

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um and this whole system has passed

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Environmental Testing so um one thing

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about microscopes and also uh you know

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fluid fluidix has a fair bit of space

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flight Heritage but in general Hardware

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uh can break and I'm sure everyone who's

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worked in a lab is dealt with a lot of

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like issues before so you want to make

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sure these things are robust and you can

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actually get it to where you want to

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launch it to

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um the optic subsystem is a nice small

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compact box and so um this is a 10

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centimeter by 10 centimeter by 13

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centimeter Cube I have uh an image on my

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phone that I didn't upload to the

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presentation because the image quality

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is not that good but for context that's

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about as tall as a banana which is very

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small for like a benchtop fluorescence

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microscope um there is like fancier

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microscopes can sometimes be like the

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size of of a person if you're trying to

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do crazy like two Photon lifetime

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Imaging or something but even an Epi

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fluorescent scope would probably occupy

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like this Podium normally and there'd be

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a lot of like miscellaneous kind of

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assorted parts so it's a really nice

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little system

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and so that contains all of the

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excitation LEDs emission filters all the

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Optics

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um and so uh this table is just kind of

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going through what the different uh bits

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of Hardware that we have are and

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um to kind of like give the the the high

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level Summary of why this is relevant

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um basically we have three LEDs that are

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common excitation wavelengths for

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different fluorescent stains that can

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also excite different types of

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autofluorescence and then

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um one thing that's actually relatively

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unique to this microscope certainly no

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microscopes I've worked with previously

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have had this is the 275 nanometer LED

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so that's actually fairly far in the

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ultraviolet range

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um and that's really good at exciting

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protein fluorescence so uh tryptophan

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phenylalanine the various aromatic amino

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acid side chains actually fluoresce

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quite brightly in the ultraviolet and so

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that can help image native fluorescent

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of samples and also excite the

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fluorescence of many different kinds of

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minerals

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um

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the objective is also set up to help uh

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well it's

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robust a space flight but also the part

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that I think is super fun is that it

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most objectives actually can't collect

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light in that ultraviolet range

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um so it is a custom objective that's

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built for those sorts of experiments

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um it has a Piezo stage that lets us do

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automated Z stacking and this is also

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been subjected to environmental and

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shock and vibrational testing and also

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radiation testing

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um to the specifications of the Europa

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Lander mission

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so um we're now on our ongoing work is

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actually characterizing this microscope

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by Imaging different types of samples to

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see what the the the limits are of what

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we can detect so um this sample is just

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kind of showing uh some of the bright

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some bright field images from a sample

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that was collected from Lake undersea in

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Antarctica and so um I'm showing here

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the 10 Micron and 1.2 Micron filters to

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kind of highlight that um we can image

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the the objects at

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um of very different sizes and you can

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see the physical uh size of the little

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uh holes there

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um and so if we were to just put this

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all onto one filter that microbial matte

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looking thing on the the

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your left

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um would probably obscure some of the

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smaller objects that we're seeing in the

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other filter sizes

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um

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and so

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um as I mentioned earlier we can also

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excite native fluorescence and so we can

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correlate this as well with the bright

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field images so we can take an image of

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an object say oh hey that looks

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interesting I wonder what that is look

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at the you know native fluorescence in

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different ranges and get a sense of okay

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wait we've got a bunch of like

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Organics that are fluorescing in the UV

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range that are all localized on this

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object that's pretty exciting

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and then we can also then stain the

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sample with fluorescent stains to look

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for more specific chemical signatures

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and so in this case I'm showing the same

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sample after it's been treated with a

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stain for primary amines and another one

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that stains lipids and so we can look

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for both the presence of those molecules

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but also their co-localization in space

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and this is a really like unique and

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Powerful kind of data that you can get

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from the microscope is not just like

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what compounds are there but seeing that

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they're associating with each other kind

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of gives you a good piece of evidence

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that you might be seeing some

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disequilibrium and some living

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um microscopy data is also

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large

286
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types of data that we've talked about in

287
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this conference so far but certainly

288
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large enough that if you were trying to

289
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send all of this data back from Europa

290
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or Enceladus it would be more

291
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complicated to handle and so one of the

292
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things that we're able to do on this

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microscope is actually take a z-stack of

294
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of images and compress it down to a

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00:11:53,990 --> 00:11:51,240
single image that's much less in data

296
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volume and so the image I'm showing

297
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right now is

298
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um an image set of images that were

299
00:12:02,870 --> 00:12:00,360
automatically collected at different

300
00:12:04,250 --> 00:12:02,880
focal planes in the sample

301
00:12:06,290 --> 00:12:04,260
um and then after it's run through the

302
00:12:08,690 --> 00:12:06,300
compression routine you end up with a

303
00:12:11,750 --> 00:12:08,700
single in Focus image and you're now

304
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suddenly gone from megabytes of data to

305
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kilobytes of data

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um and then

307
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um you can even go further with that and

308
00:12:24,350 --> 00:12:22,560
you'll notice that this sample has uh

309
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bright objects against a dark background

310
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we don't actually care what the like

311
00:12:30,829 --> 00:12:28,980
Photon shot noise is on the background

312
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for our purposes that might as well just

313
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all be zero and so the final compression

314
00:12:37,910 --> 00:12:35,279
step is actually taking that and setting

315
00:12:40,130 --> 00:12:37,920
it to zero and so we're just recording

316
00:12:42,650 --> 00:12:40,140
um the actual bright objects that are

317
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detected in the image and so that all

318
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reduces the data size quite

319
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substantially

320
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um without really getting rid of any of

321
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the information that we actually want to

322
00:12:56,150 --> 00:12:54,889
um so how would this work in in a

323
00:12:57,290 --> 00:12:56,160
mission I just talked about a lot of

324
00:12:59,449 --> 00:12:57,300
different parts and now I'm going to

325
00:13:01,670 --> 00:12:59,459
bring it all back together into how this

326
00:13:05,509 --> 00:13:01,680
system could be implemented

327
00:13:07,069 --> 00:13:05,519
um so and I have the little Europa

328
00:13:10,129 --> 00:13:07,079
Lander picture there

329
00:13:12,230 --> 00:13:10,139
um but basically uh we pretend that the

330
00:13:14,930 --> 00:13:12,240
microscope is somewhere in that Lander

331
00:13:17,509 --> 00:13:14,940
um you would take your sample process it

332
00:13:19,670 --> 00:13:17,519
pump it through the fluidic system you

333
00:13:22,310 --> 00:13:19,680
can image it on the microscope you're

334
00:13:25,310 --> 00:13:22,320
going to end up with megabytes of data

335
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that you can then compress back down to

336
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your kilobytes of data send it back to

337
00:13:32,509 --> 00:13:30,300
your ground-based Observer that will

338
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then be able to say oh okay cool we got

339
00:13:36,050 --> 00:13:34,620
a microbial mat on Europa time to hit

340
00:13:38,210 --> 00:13:36,060
the Press

341
00:13:42,350 --> 00:13:40,910
and so uh yeah just briefly

342
00:13:45,050 --> 00:13:42,360
acknowledgments there's been a lot of

343
00:13:47,329 --> 00:13:45,060
funding that's gone into this

344
00:14:02,150 --> 00:13:47,339
um and um I'll be happy to take any

345
00:14:06,710 --> 00:14:04,730
great talk uh this is Chad pazoriski at

346
00:14:08,569 --> 00:14:06,720
Georgia Tech so

347
00:14:10,610 --> 00:14:08,579
um I'm really happy that you touched on

348
00:14:12,350 --> 00:14:10,620
uh compression first of all uh of the

349
00:14:13,850 --> 00:14:12,360
data because that's incredibly important

350
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and

351
00:14:18,050 --> 00:14:16,019
um I was also really thrilled that you

352
00:14:19,310 --> 00:14:18,060
talked about the um sample prep and

353
00:14:20,509 --> 00:14:19,320
fluidics because I think that's

354
00:14:23,870 --> 00:14:20,519
something that a lot of people don't

355
00:14:26,329 --> 00:14:23,880
consider for these NCG missions uh my

356
00:14:29,389 --> 00:14:26,339
question has to do with

357
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um comparing uh microscopy and staining

358
00:14:35,389 --> 00:14:33,000
with uh perhaps something like Raman or

359
00:14:37,009 --> 00:14:35,399
IR where you could get information about

360
00:14:38,870 --> 00:14:37,019
at least functional groups of the

361
00:14:41,569 --> 00:14:38,880
Organics present and co-locate things

362
00:14:44,030 --> 00:14:41,579
that I imagine similar resolution

363
00:14:46,610 --> 00:14:44,040
spatially but you wouldn't need to

364
00:14:49,189 --> 00:14:46,620
include those processing steps so would

365
00:14:51,410 --> 00:14:49,199
you advocate for a system that's I guess

366
00:14:52,910 --> 00:14:51,420
maybe not redundant but but has both of

367
00:14:54,829 --> 00:14:52,920
those features or do you think one is

368
00:14:56,210 --> 00:14:54,839
better than the other yes this is

369
00:14:57,829 --> 00:14:56,220
actually a super interesting discussion

370
00:14:59,990 --> 00:14:57,839
and we can definitely also talk more

371
00:15:01,850 --> 00:15:00,000
about it at lunch but the the high level

372
00:15:03,610 --> 00:15:01,860
thing that I'm going to say is that um

373
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you you get

374
00:15:08,389 --> 00:15:07,019
you're going to have different pros and

375
00:15:09,910 --> 00:15:08,399
cons and get different types of data

376
00:15:12,829 --> 00:15:09,920
those techniques so if you're doing

377
00:15:15,949 --> 00:15:12,839
let's if you want to get spatially

378
00:15:17,269 --> 00:15:15,959
correlated Ramen data when you're

379
00:15:20,449 --> 00:15:17,279
building a microscope and you're trying

380
00:15:21,829 --> 00:15:20,459
to image something unless you're using a

381
00:15:25,189 --> 00:15:21,839
super resolution technique you're

382
00:15:28,310 --> 00:15:25,199
limited by diffraction in terms of like

383
00:15:29,870 --> 00:15:28,320
the the resolution that you can get and

384
00:15:31,490 --> 00:15:29,880
that is proportional to the wavelength

385
00:15:34,850 --> 00:15:31,500
that you're using so when you start to

386
00:15:36,470 --> 00:15:34,860
get into like the IR you know range and

387
00:15:39,590 --> 00:15:36,480
things like that

388
00:15:41,930 --> 00:15:39,600
um your spatial resolution starts to be

389
00:15:44,930 --> 00:15:41,940
more limited than it is in the optical

390
00:15:47,090 --> 00:15:44,940
range the flip side is you're right you

391
00:15:48,230 --> 00:15:47,100
can get precise chemical information out

392
00:15:51,590 --> 00:15:48,240
without

393
00:15:55,670 --> 00:15:51,600
um you know adding labels and so I think

394
00:15:57,590 --> 00:15:55,680
that definitely those are uh like

395
00:15:59,930 --> 00:15:57,600
in in in in the perfect mission

396
00:16:04,189 --> 00:15:59,940
architecture I think you do both

397
00:16:05,750 --> 00:16:04,199
um but you know and um unfortunate you

398
00:16:07,250 --> 00:16:05,760
know it's it's it's unfortunate that we

399
00:16:08,930 --> 00:16:07,260
can't send all the instruments that we

400
00:16:11,090 --> 00:16:08,940
want out there but like in my ideal

401
00:16:13,129 --> 00:16:11,100
world I think those that would be a

402
00:16:14,090 --> 00:16:13,139
powerful like correlative technique

403
00:16:17,090 --> 00:16:14,100
there

404
00:16:19,250 --> 00:16:17,100
um to kind of get a a strong sense of

405
00:16:25,069 --> 00:16:19,260
what you're seeing in your sample

406
00:16:25,079 --> 00:16:33,610
foreign

407
00:16:38,930 --> 00:16:37,069
again thanks for an awesome talk this is

408
00:16:40,850 --> 00:16:38,940
really exciting so I was wondering what

409
00:16:45,110 --> 00:16:40,860
sample prep looks like

410
00:16:46,610 --> 00:16:45,120
um in the Rover or whatever and um what

411
00:16:47,810 --> 00:16:46,620
the capabilities are for that so could

412
00:16:49,249 --> 00:16:47,820
you actually do like incubation

413
00:16:50,810 --> 00:16:49,259
experiments and then look at it with

414
00:16:52,069 --> 00:16:50,820
microscopy or is it coming straight from

415
00:16:53,150 --> 00:16:52,079
the environment going straight to the

416
00:16:56,870 --> 00:16:53,160
microscope

417
00:16:59,990 --> 00:16:56,880
yeah so um it's the

418
00:17:02,090 --> 00:17:00,000
the the design that we had in mind for

419
00:17:03,889 --> 00:17:02,100
for sample prep here and when I said I

420
00:17:06,230 --> 00:17:03,899
say we it's not

421
00:17:08,030 --> 00:17:06,240
um us this was kind of the built around

422
00:17:10,309 --> 00:17:08,040
the Europa Lander specifications but

423
00:17:12,169 --> 00:17:10,319
essentially you'd have a sample cup that

424
00:17:15,409 --> 00:17:12,179
would collect an ice sample and melt it

425
00:17:18,409 --> 00:17:15,419
down and so that was kind of the um

426
00:17:20,870 --> 00:17:18,419
the initial like thoughts behind um the

427
00:17:22,309 --> 00:17:20,880
design here but the fluidics processing

428
00:17:23,750 --> 00:17:22,319
system can basically handle anything

429
00:17:25,069 --> 00:17:23,760
that's in a liquid state but the

430
00:17:28,909 --> 00:17:25,079
assumption would be that it's a

431
00:17:30,549 --> 00:17:28,919
relatively unprocessed field sample

432
00:17:33,230 --> 00:17:30,559
um

433
00:17:34,730 --> 00:17:33,240
relatively unprocessed because obviously

434
00:17:36,110 --> 00:17:34,740
if it's like ice or something and you're

435
00:17:38,090 --> 00:17:36,120
melting it down you are still doing

436
00:17:39,590 --> 00:17:38,100
something to that sample

437
00:17:42,950 --> 00:17:39,600
um

438
00:17:42,960 --> 00:17:46,610
okay we have time for one more question

439
00:17:51,110 --> 00:17:49,850
hi thank you for the great talk

440
00:17:53,990 --> 00:17:51,120
um

441
00:17:57,110 --> 00:17:54,000
my question is uh maybe I missed that

442
00:18:01,010 --> 00:17:57,120
you mentioned how

443
00:18:04,250 --> 00:18:01,020
are you going to differentiate uh

444
00:18:04,970 --> 00:18:04,260
any other particles

445
00:18:08,930 --> 00:18:04,980
um

446
00:18:13,210 --> 00:18:08,940
from the living materials like can you

447
00:18:16,010 --> 00:18:13,220
specifically stain genetic material or

448
00:18:17,810 --> 00:18:16,020
anything biological

449
00:18:19,310 --> 00:18:17,820
yeah so that's actually that's a great

450
00:18:23,090 --> 00:18:19,320
question and

451
00:18:24,230 --> 00:18:23,100
um so the the the the goal here is to

452
00:18:27,110 --> 00:18:24,240
kind of pick up as many different

453
00:18:29,690 --> 00:18:27,120
breadcrumbs as you as as we can to try

454
00:18:31,130 --> 00:18:29,700
and uh differentiate and so you

455
00:18:32,630 --> 00:18:31,140
mentioned stains for genetic material

456
00:18:34,070 --> 00:18:32,640
one of the stains that we would be

457
00:18:36,049 --> 00:18:34,080
launching with this is a stain for

458
00:18:40,190 --> 00:18:36,059
nucleic acids

459
00:18:43,370 --> 00:18:40,200
um now obviously that would be really

460
00:18:44,870 --> 00:18:43,380
awesome if we found DNA on Europa or

461
00:18:46,549 --> 00:18:44,880
Enceladus but

462
00:18:48,289 --> 00:18:46,559
um there's it's kind of an open question

463
00:18:49,970 --> 00:18:48,299
whether we would

464
00:18:52,610 --> 00:18:49,980
um and

465
00:18:54,590 --> 00:18:52,620
um so the idea behind looking for the

466
00:18:57,289 --> 00:18:54,600
something like the co-localization of

467
00:19:00,950 --> 00:18:57,299
amines and lipids is that that gives you

468
00:19:02,330 --> 00:19:00,960
a more like agnostic idea of like hey

469
00:19:04,070 --> 00:19:02,340
that might be living material because

470
00:19:06,470 --> 00:19:04,080
you've got these you know chemical

471
00:19:09,409 --> 00:19:06,480
signatures that are co-locating

472
00:19:10,610 --> 00:19:09,419
um the other thing is um so one of our

473
00:19:12,529 --> 00:19:10,620
um collaborators actually the person who

474
00:19:14,890 --> 00:19:12,539
helped build the microscope

475
00:19:17,810 --> 00:19:14,900
um is has some ongoing work

476
00:19:20,630 --> 00:19:17,820
characterizing the fluorescence of uh

477
00:19:22,130 --> 00:19:20,640
diff of a wide library of minerals to

478
00:19:23,870 --> 00:19:22,140
get a sense of like what they would

479
00:19:26,650 --> 00:19:23,880
potentially look like

480
00:19:29,690 --> 00:19:26,660
um on the the microscope so we can

481
00:19:30,830 --> 00:19:29,700
hopefully differentiate between that and

482
00:19:32,630 --> 00:19:30,840
things that might be a little bit more

483
00:19:35,750 --> 00:19:32,640
biological but spectroscopically and

484
00:19:37,190 --> 00:19:35,760
then also you know morphologically

485
00:19:40,190 --> 00:19:37,200
um

486
00:19:45,900 --> 00:19:40,200
yeah a great question