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[Music]

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[Applause]

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thank you it's an opportunity to present

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Antony today Antony is based on

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primarily microscope that's an atomic

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force microscope it was named in honor

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of the father of microbiology Anthony

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and Meagan Leeuwenhoek I the clever

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naming for that as due to brent chris

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iners Hey midnite acronym searching

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algorithm uh-huh the I'm gonna just get

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started in here okay so I as you know

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one of the primary goals of the Europe

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Lander science definition team was

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address to address characterizing bio

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signatures in Europe as near subsurface

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materials there are a couple of

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objectives that we address with a

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microscope the first is to identify and

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characterize morphological and/or

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textural features of Europe this near

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subsurface and the investigation that

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Anthony accomplishes here is to

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characterize particulates and samples

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that are 0.2 microns to 1500 microns in

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scale the second objective the antony

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addresses is to assess the structural

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and compositional nature of europe is

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near subsurface materials there are two

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investigations here one of them is to

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measure the mechanical properties of the

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sample and the second is to potentially

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determine the composition of

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particulates afm you may be familiar

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with it's been around for about 40 years

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works is a pretty simple instrument and

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it works by a cantilever interacting

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with a sample it raster's over the

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sample and it creates a

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three-dimensional topographic map if

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Emma's tip is often used in material

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science in the semiconductor industry

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but can also be used to detect biology

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particulates such as this cell were

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imaged in a 1999 paper if this came from

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Lake Vostok accretion ice you can get

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really high-resolution image at the sort

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of sub sub cell level even looking at

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textures that are on the cell surface

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but in addition to getting a 3d

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topographical map you

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also get nanomechanical properties

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things such as the sample stiffness the

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dissipation of energy when the

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cantilever interacts with the surface

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and adhesive properties and so you can

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see how sort of a diagram of the cartoon

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and the top figure here on the bottom is

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a scale showing the stiffness as a

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as measured as kiloPascals over eight

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orders of magnitude different between

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cells and hard materials like steel so

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AFM bio signature detection technology

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as we have thought about it builds on

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spaceflight heritage there are a number

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of assets to using AFM in space in

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addition to its submicron 3d resolution

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and the potential for nano mechanical

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property determinations the these

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instruments are lightweight they have a

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small footprint and low power

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consumption they can operate an air

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liquid and in vacuum the they've also

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been flown recently in space on two

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different missions one of them was flown

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on the Phoenix mission and the other on

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the Rosetta mission in both cases they

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imaged particulates on Mars on the

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Phoenix mission in particulates size

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distribution and soils on the Phoenix

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mint and the rosetta mission they

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measured cometary dust and an observed

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large particle aggregates mark Bentley

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wrote a paper post the Rosetta mission

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that was really on lessons learned from

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operating AFM's in space which we have

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adopted into the design going forward

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and thinking about antony some of these

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are co registering if you use a no

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register ring an optical microscope

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image with the atomic force microscope

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image including an array of cantilevers

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that have different spring constants in

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different tip morphologies and then

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being able to operate the instrument

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both in open and closed loop formats

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which they was required on the Rosetta

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mission so one of the things that I

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think AFM has a real advantage to is

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detecting really small bio signatures we

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don't know to what to expect and going

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to the surface of Europa

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and we didn't know what to expect when

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we went to Lake Vita which is a lake in

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East Antarctica and the like and McMurdo

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Dry Valleys where the sand plan I don't

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know that you can see very well from the

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back that you can see they're really

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bright images here these these are what

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I would call kind of normal-sized

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bacterial cells and this background sort

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of starry night barely shows up when

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you're looking at really what we have is

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like one of the brightest DNA stains

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under epi fluorescent microbes

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microscope if we look using an AFM you

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can see really really what these what

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these particles look like you can zoom

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in at higher resolution and and start

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looking at the surface textures of the

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cells so we have also been interested in

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can asking the question can we discern

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bio signatures from abiotic particles

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using AFM based on their nano mechanical

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properties this is a image that we

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collected where the we we mixed silica

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beads so if I go back here so the round

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features which you may or may not be

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able to see our silica beads that we

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mixed with bacterial cells and if we

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look at the forest curves from these

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particles we can see that this is on a

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glass slide so the black dots are the

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are plotted here on the graph the glass

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background is not different from the

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glass beads that are in blue and we have

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particulate organic matter that is as

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Roger calls the schmutz from the cells

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that is in green this is softer and then

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even softer yet are the cells that are

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in the and the figure we did a second

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kind of similar experiment but in this

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case due to a conversation we were

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inspired that we had with Mitch Schulte

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a few months ago he asked us a question

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could you actually discern the

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difference between carbonaceous

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chondrite meteorite and bio and cells

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and so we obtained some i-n

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crushed powdered meteorite sample we

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screened it through a 5 micron sieve and

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mixed it with three different bacterial

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cell types in this case the meteorite

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which is here in the particle in the

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middle of this aggregate of cells shows

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up to be quite stiffer not as stiff as

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the background of glass and then the

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cells have different they are softer yet

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and they have different curves for the

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different types of cells that we had in

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in this case we mixed an Antarctic

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bacterium called pseudo Vibrio with

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bacillus and s worse in equine so we

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think we can discern particles of

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different types of properties from from

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life you could imagine then that if we

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went and got a library of different

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types of cells and particles that we

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might be able to just separate these in

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in three dimensions or in here in two

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dimensions on a on a graph based on

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their biophysical properties as Peter

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talked about a little bit in his talk

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but identifying morphological features

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of particulates and I see potentially

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salty samples is going to require sample

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processing and along these lines we

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partnered with the Monterey Bay Aquarium

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Research Institute to develop a concept

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for this instrument which would be a

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sample handling system Ambari has a lot

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of history and experience in operating

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remote autonomous platforms in the ocean

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on earth this is the 2g ESP that was

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originally funded by a NASA a step

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project with the the concept they would

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evolve this design into space based

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operations and the - 2g ESP here is very

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large but this is actually a functional

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molecular biology platform where it's

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not only processing collecting samples

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and analyzing them but also doing real

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molecular biology DNA extractions PCR

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and and remotely communicating that

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information back to the lab they use

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titanium pucks which are inert for

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sample processing and

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this system is is again complete and

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remotely operated they have a new system

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that they have integrated into this

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submersible which is based on

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microfluidic cartridges which has a it

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rotates around and as actuator driven

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inside the head of the submersible so we

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have adopted elements from from both of

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these designs into the concept for

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Antony in which the titanium puck would

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be delivered to the instrument the

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central processing unit here then would

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accept the sample and process take it

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pressurize the sample in order to be

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able to melt it without adding heat or

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very much heat and then it would be

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passed through different screen sizes

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and then those samples can then be

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rotated and passed by an optical

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microscope an afm and potentially a

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third instrument that is on the lander

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this all the design fits in the ten

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centimeter by 10 centimeter space that

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was described in the PIP so there are

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several elements that even though we're

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basing this on the heritage design of

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what has flown before there are a couple

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of upgrades that we can do 20 years

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later than when these developments were

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made the so piezo resistive probes can

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be upgraded as well as the actuator and

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fine XY scan range to move the sample we

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think calibration targets are needed for

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every sample processed so that you can

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get tip shape on each sample as well as

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the XYZ and mechanical force scales

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autonomous impressions are going to

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require image analysis some of this can

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be adopted from heritage algorithms that

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were developed from the Phoenix mission

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but we also have ideas for how to

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compress the data to preserve the high

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information aspects of it as well as for

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developing neural networks for particle

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classification this is based on work

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that was done at Oakridge and published

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several years ago with a project

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collaborator Steven Jesse we also have

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there's preliminary work that has been

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done with where you can use the AFM to

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do correlative mass spectrometry with

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particles by heating up the tip of the

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AFM you can volatilize the sample and

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bring it into a mass spectrometer and so

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this could potentially really give us a

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lot of information about particles being

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observed the advantages of Antony then

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are that this is a system as we've

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designed it has flexible architecture we

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can partner with other instruments it's

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integrated sample preparation system

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affords us to have high resolution image

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and proper image and property

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determinations the topographic maps will

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be useful information regardless of

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whether the particles are biogenic or

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abiotic in nature and the Nano

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mechanical forces can potentially help

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us inform between life-forms and and

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those that are that are not life in

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terms of stiffness adhesion dissipation

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of energy and even in the absence of

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life I think that this kind of a

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platform can yield invaluable

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information about microscope materials

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and features in the near subsurface of

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Europa the the team here is large and

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great and I have learned a ton from

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working with all of them Roger approach

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is sort of the mastermind behind the AFM

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design crystal and is the chief and

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president of Ambari and with Jim birch

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and Doug market have designed the Europa

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generation ESP thank you

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yeah given that you're looking at very

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tiny scales with AFM how do you deal

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with searching for things that are

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extremely low concentration that you're

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likely to see 100 per milliliter or

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something like that see if the search

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throw a big haystack refer so we have an

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the the concept is that we would

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concentrate the sample onto a pretty

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into a pretty small surface but also

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that we would first inspect the the

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sample with an optical microscope and

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you have a larger scale and then be able

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to identify the high entropy areas of

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that image and then localize the AFM to

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where to go do the work and you can even

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still do nested scans like they've done

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on both of the other missions with AFM

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so you can start at larger and then go

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in and a higher resolution at a smaller

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area another way of distinguishing

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between whether you're looking at

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inorganic material or organic or bio

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material would be to do a tip enhance

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spectroscopy such as tip enhanced Raman

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or chip enhanced infrared and I know

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it's probably at a lower TRL but what do

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you think about the possibilities of

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eventually moving to such an enhancement

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of the technology yeah I think that's a

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super super super interesting an area to

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develop for the future and to get that

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kind of instrumentation into the sort of