Us last night.

It's Thank you very much.

It's a great pleasure and an honor
to be here in particularly if you're

given invited seminar and all you have to
do is carry your laptop across the street.

That makes logistics a lot easier.

The title of my presentation
is Jeff has mentioned already

will indicate separated that we're going
to try to span actually breach between

systems biology and
what we call systems in a little X.

and some of you might ask
the question what is systems and

a lifting so I've never heard about
this expression in particular.

Considering that it's already hard
to define what the systems biology.

So what we're going to try to do today.

It goes back to about two thousand and two
where I got invited to talk a little bit

about the role that analytical chemistry
complained the context of biology and

that talk was given at
a conference called Systems X. and

that was when Switzerland started actually
their systems biology initiative from

the very large scale and they invited a
couple of people just to present thoughts

on what they think is system biology and
what that the other disciplines that

are needed actually to feed into
these very large scale initiatives so

they will try to propose an area we're
going to call systems analytics and

I'm very fortunate that I can recap
what happened in the last four or

five years to substantiate this field and
hopefully convince you that systems in

the litigious is something that will
be complementary to Systems Biology.

So a lot of the research we do in the lab.

I'm going to focus on a couple of areas
which should illustrate why we think

that analytical chemistry will play
a major role in Lucid a thing.

Some of these complex process
is associated with biological

systems we're going to look at any area.

Called multi-functional scanning Propes
and you saw on the title SLATTER read it I

have a dear collaborator on that
Dr Christine crams who's responsible for

the development of this
multi-functional skin in Propes and

I hope to be able to convince a dead by

trying to integrate analytical
techniques into a generic platform.

We will be able to actually see more even
though we are interested in smaller and

smaller dimensions on the spatial scale
on a temporal scale on a volume scale.

If we go down to smaller biological
units and at the very end.

I'll show you probably spend in
the last fifteen minutes and

sort of an outlook on emerging
technologies that we see coming up in

the next couple of years where we can even
expand what this multi-functional scanning

probes do by spectroscopic abilities
in particular infrared spectroscopy and

a long way from length range to far
infrared and Terra spectroscopy.

So forgive me if as an invalid
tickle chemist I will try to define

what these systems biology and I know
there are many many definitions out there.

So I'll just throw out there something
that we as analytical chemistry

is important and
we really feel that Systems Biology

integrates does this sort of concerted
approach you're really concerned with

a couple of disciplines that you're trying
to pull together actually to answer

this complex prayer questions
that we're facing in terms of.

The processes that are going on in
living biological entities which can be

as large as the entire organisms or

as small as actually single cells and
trying to elude what's going on there.

And of course if you look at this
biological entities there are some

multiple cellular processes that we
can think of which can range from

transport and trafficking of molecules
which the reader present already in

the cell which we artificially trying
to introduce to the cell by a Bridewell

riot you have mechanisms involving
basically a lot of chemical actions and

interactions at the cell surface but
also within the cell.

So what we when we were asked.

Actually to get our thoughts and then a
little chemistry we said well this sounds

like a very systematic approach
that's already inherent in this name.

So do we have all the analytical
tools already available to probe

all these processes in the same
systematic fashion which means can we

look at the transport traffic in for
instance of a molecule from it and

then make point of view by measuring it
to quantitatively and qualitatively and

observing rather it's doing it to sell
surface how it's interacting for instance

with receptor is how it's interacting
with the interiors of this cell.

And so what we basically would like to
propose now and at the end we're going to

get back to hopefully it definition of
what we think now those systems and

allude to maybe is we would like to answer
actually first of all rich species out

there and that with very high molecular
selective U.-T. and sensitivity.

We want to know actually how much is there
said dealy all the analysis we're going to

do at this biological surface or
interface should be quantitative and

ideally we want to be able to observe
how the system is changing or

how this process is a changing with time.

So we want to be able to do to
start a sexually kinetically or

dynamically So what does that entail.

We hope that actually analytical
chemistry can assist system biology in

delivering actually the data that you use
for all your models at a molecular level.

So we would like to provide
it was quantities and

qualification of the species
that are present and

involved in this process is and in fact
provide true experimental evidence for

all the modeling all the prediction
that is done in particular if you think

about by informatics and by a mathematical
assisting actually systems biology.

So truly what we'd like to do is fill
libraries with numbers with real numbers

that we've measured at this biological
surfaces or interfaces and being able to

assign these numbers actually to
the actual processes that are going on.

So.

Let's think a little bit and again from
and then a little point of view we view

actually a cell or a biological specimen
or a logical unit if you would like to

investigate as an analytical compartment
and I'll get back to that in the mean it.

What I mean by analytical compartment it's
an enclosed space which is interacting

with its environment and we would like
to probe actually this interaction.

And if we are logically
trying to divide up

which techniques do we have available
to probe these interactions.

We can largely say we have a lot of of
course invasive analytical techniques mean

meaning that we have to do a sample
preparation we might have to

homogenize our cell lives our cell and
then try to analyze basically

what's left with a variety of tick tick
needs I'm not going to go into detail and

then we have what we would
call noninvasive for

let's better call them same invasive
techniques because this is actually very

few techniques where we can only under
codes observe what our biological entity

is doing without actually
disturbing the process is going on.

So let's call this process the sort
of these techniques Amy and

basically we're trying to observe what's
happening with our biological entity.

So if we take and
this is by no means an exclusive list or

if we take these types of
analytical technologies.

There's one thing that's common
to these techniques mostly if

we're trying to apply a couple
of these techniques to

learn something about
the biological environment.

We usually collect our
information in sequence.

So we apply one method to progress certain
process or to do an electro chemical

experiment then we might with
a mass spectroscopic experiment or

an offical experiment so usually
we're going to sequence surely prope

the molecular space that is present
within or at this biological surface.

So why is this a problem from
an analytical point of view.

Again I'm not the biologists provide you
with the analytical point of view where

we think the challenges are in probing
these biological systems with all

these beautiful analytical taken.

Yes we consider now our biological
specimen is what I would

call a molecularly densely packed
analytical compartments we have many many

species present actually at
a very very small location or

within the very small volume then already
that poses a certain challenge to our

analytical techniques if you look at
individual cells couple of microns

a couple of tens of microns we have
actually a fairly small compartment and

we're trying to probe with
molecular level sensitivity and

selectively three certain
constituents present within or

interacting with this compartment
these systems are frequently changing.

If we look at life biological specimen.

We're going to have a permanent
change of topic around matrix

that we have to account for.

We still want to observe maybe only one or
two molecules.

But we have a tremendous background
that causes a fluctuating signal

that we have to account for

actually practically It's a rare that
we're interested only in one species.

Even though the examples I'm gonna keep
you today rutile with only one species but

practically we would like
to be able to follow and

trace multiple constituents
interacting in this biological domain.

We would like to do that quantitatively
which means not only what is there but

also how much is there and we would like
to do that actually ideally with temporal

and letter a resolution so here we are
basically proposing a wish list on what

all these and a little technique should
do for us at a live logical surface or

at a live biological interface.

So.

The way we develop this idea
of systems and it ticks is

to take actually these requirements
which is sort of the ideal situation for

us if we observe to spot
a logical units and

try to develop these techniques to move
from the sequential information gathering

process to what we would call a simple
train as information gathering process so

ideally we would like to probe all that
at the same time at dealing with the.

Same temporal and
spatial resolution and being able to

answer actually it is complex questions on
this process is at the logical surfaces.

So the solution we have proposed
a couple of years ago to this problem or

one possible solution is an approach
we would call multi-functional

analytics It means we have to integrate
multiple of these techniques to

operate in a space in time correlated
fashion ideally synchronized

at this last biological interfaces and
provide us hopefully with

noninvasive information which is
interiorly this time and space correlated.

And today I'm going to give you
a couple of examples of such

instrumental techniques which allow us
actually to probe multiple parameters

at the same time with this inherent
time and space correlation.

So instead of focusing in details
on the biological systems

that we've investigated so
far I'm going to focus more on

the end a little approach that we fused
into technologies we've developed because

I think that's going to lead us actually
to this exciting future perspective

of integrating a number of analytical
techniques into such a platform.

Nonetheless we will have to look at
some examples and in fact if we proposed

this integration of analytical
techniques onto a unified platform.

It means that we go actually beyond
integration of technologies we have to

integrate these technologies with
the systems that we're interested in and

in fact I really emphasize this work
here a sensitive integration and

that's a lesson we've learned over
the last ten years working with medical

partners at Emory University and the Johns
Hopkins Hospital that really sensitive

means to understand the problems and
the analytical questions that biology and

medicine really would like to answer so
a lot of our technology development

is really guided by the questions
that biology would like to answer.

And we hope actually.

That we can provide the appropriate
information on it and then make scale

to assist actually in addressing
the complexity of these systems and

I'm going to give you one example of
actually one of the key an alliance we've

been concerned with over the last seven
years a comparatively small molecule and

this small a small molecule is A.T.P.
at and it's in triphosphate And

the reason why we focused actually on to
this molecule is that amazingly enough

even though there's a couple of analytical
techniques established since twenty thirty

years to measure A.T.P. it's still a
challenge to measure A.T.P. actually with

high a temporal and spatial resolution at
the surface of life biological specimen.

Why have we focused on A.T.P.
infective focused on A.T.P.

because there are two systems that
are interested in which involve A.T.P.

sort of as a main molecular
player involved in

in particular signaling process is at
the surface of this biological specimen

on the left side here we see an example
of a collaboration we have with.

Johns Hopkins Hospital.

This is actually a car
wrote that body preparation

carolled that body that is responsible for
controlling our respective Torrie

processes sits at the bottom
occasion of the carotid arteries.

It's a sellout sambal of about two
three hundred microns in diameter and

this cell are solidly safe and

tested sensory organ actually it measures
the concentration of C O two in blood.

It measures to concentration of oxygen and

blocks in there by assists us
in controlling Torrie processes.

It turns out that the Met to ration
off this current that body and

whether that happens under oxygen
tension on the Oxygen stress may

affect actually the later development
office period Tori diseases and

that's what collaborated
Johns Hopkins is investigating.

Interestingly enough in this whole
signaling cascade that the current

IT body uses to.

Regulate this process is A.T.P.
is one of the key molecules

that actually interacts with receptor
said the surface of this corroded body.

The second system that were interested
in this is a five years and

each program that we've developed together
with Emory University with the school of

cell physiology where we are looking
actually at lung Epi filial cell surfaces

and in particular the molecular
conditions during the development of

cystic fibrosis where it turns out that
again to cystic fibrosis trans membrane

regulated here expressed at the surface
of the separate cells utilise this or

it gets triggered by a T P So
A.T.P.'s again a key

molecule in this molecular level process
is associated with cystic fibrosis.

The problem actually in both
cases is to measure A.T.P.

with very high lateral resolution
at the surface of this live.

Logical specimen.

So the larger context actually
of why we would like to do that

is that they gained the respect of Tory's
system is for us to send a little chemist

a very interesting example because
it allows the body to interact

directly with parameters from
the external environment.

So again we're talking about the corroded
part the which controls are a spirit or

a function but at the same time as
soon as we actually inhale air and

get down into this A.B. all the space.

We have a direct interaction of whatever
is present within the same band

environment at the surface or at
the inner liner basically it is epi feel

a cell layer which separates the inside
of our body from the outside world.

So we're really interested in the
processes that are occurring directly at

this biological interface and the reason
why we're interested in that is that we

believe that if we understand these
processes at the fundamental molecular

level then we might in
future actually get.

To a level of understanding that
allows us to trace actually these

processes in the tickler to
trace processes where D.S.

molecular interactions are disturbed
like during disease cases

pulmonary disease cases we might
be able to trace these processes.

But ultimately looking only at
the molecular composition of for

instance a breath or breath calm and say.

But I can tell you right away that we're
probably fifteen years away from there

because it's going to take at
least that long that we understand

these processes down here
at this cell boundaries.

Before we can even think about which
panels of biomarkers we can try to

trace and use actually as
prognostic patterns in breasts and.

So let's look at A.T.P. real quickly and
again I'm not going to go into details

on that to be able to provide you with
the speaker picture we have in mind but

I'm happy to answer any more
detailed questions afterwards.

In fact if you look at
the measurement of A.T.P.

a really it goes back to the eighty's
where people have developed by a census

to be able to procreate E.P. and
it's great and it works extremely well.

But there is a challenge
associated with it.

The challenge is that actually none of
these systems have really been report

reported to work quantitatively at
Lafe a logical surface to surface

is probably with the exception of Lucifer
in the city for a system which is even

a commercial keep you can buy actually for
the optical detection of A.T.P. but

it ever ages ever it is over at
the enter entire extracellular volume.

It does not allow to probe A.T.P.

for instance that might be exercised
totally released at the surface of a cell

which are events which happen at a scale
of about hundred two hundred nanometers

what we'd like to do is probe eighty P.
at a scale of about hundred or

two hundred nanometers in
the particular case of cystic fibrosis.

This is about that I mentioned all
scale where our collaborators suspect

that the C.F. T.R.
mediated expression of A.T.P..

The SO surface is happening.

So really the motivation for us was to
develop in the little platforms that allow

us to specifically address a very small
molecule in this complex environment but

it is same time being able to not only
localize it and spatially resolve it.

But also temporarily resolve
actually when this molecule

is released at the surface
of the logical entity.

So how does that work.

I'll give you a quick example on
the bio sensor technology that we're

pursuing in our laboratory which should
assist us actually in this process and

help us to very specifically pull out
the molecule we're interested in and

it's one concept which is
actually a fairly generic concept

that can be developed for
a wide variety of molecules.

The technology we're going to use as
a transducer to measure actually how much

A.T.P. is present.

It's going to be electro and

a little chemistry we're going to use an
electrode surface and then M. per metric

measurement which will correlate in terms
of the current that we're measuring it

is an electrode directly with the amount
of A.T.P. or the concentration of A.T.P.

that's present at the surface
of a logical logical system.

The electrode reaction that we're
going to use actually is going to be

a very simple reaction.

We're going to use actually to enzymes
glucose oxidase and hacks of kindness and

these to actually have
a competitive reaction for

the substrate glucose glucose
oxidase going to produce glucose and

I guess it all could come elect to own and
produce is how to temper offset how to

temper oxide is actually the Electoral Act
of species we're going to

detect at the surface of electro oxidizing
it is six hundred fifty million volts.

OK.

If we add hex so kind a is a second and
sign into the game.

What's going to happen is the Hexa kind
is if eighty piece presents going to

catalyze the reaction to glucose
six phosphate OK what does

it mean it means less glucose going to be
available to produce hydrogen peroxide.

So the.

If eighty piece present we should see our
current signal decreasing because we're

going to produce less hydrogen peroxide
to be detected at our electrode.

Again I'm not going to go into
details on the immobilisation of this

type of enzyme chemistry at the Elektra
surface but there's about sixty years of

literature out there and how to immobilize
enzymes of receptors at the surface of

a wide variety of electrode materials but
what I would like to get it is

that we can do this at a very very
small scale we can take what is called

a micro electrode those are Elektra lots
with that meters anywhere between ten and

twenty five micro meters and
now we're going to attach a whole and

some chemistry on to these very very
small electrode surface of course we will

have to have sufficient sensitivity to
measure very small Farraday current or

current drops due to the production
of the hydrogen peroxide or

reduced production of hydrogen
peroxide if A.T.P.'s present.

Again I'm not going to go into all details
on the measurement technology just to say

so much that these Elektra with a sample
is can be routinely fabricated Nowadays

you can even buy them so electrolytes down
to a diameter of about ten micro meters

a commercially available you can
also get what we showed on here.

So called dual parallel trolls
where we have two electrodes

which can be individually addressed which
are separated by this insulating glass

shielding over here.

What you might notice here is that we
modified run of these electrode surfaces

with eye and so I met a great commission
layer and yeah they electrode surface

remains actually unmodified why are we
doing that we would like to use

Actually the signal of this electrode to
position I electrode very closely a bath.

Logical surface so we want to be
close to the surface and measure for

instance in this case with the still
in some bias sensor over here A.T.P.

produced at the surface of
the spar logical system.

In fact again we're not going to
look into the details of scanning

the lecture chemical my costs go up and.

How to do that just to see so

much that we can use actually oxygen
present in our solution and the current

response of oxygen to actually move these
electrodes and see the reaction closer and

closer to our surface until we see what is
called a positive or negative feedback.

Meaning that the surface
properties here going to

influence the Farraday current
that we produce at this electoral.

That allows us to get to select
It's very close to the surface and

very close in the electro chemical terms
means a couple of Electrolux of radio

way from the surface.

Well they've already indicates to you
if we make I electrode smaller and

smaller we have to get closer and
closer to the surface to do a quantitative

measurement and
that's going to be the next challenge.

We will discuss.

I'll just show you that the system
actually works beautifully.

We have to say synthetic cell we
call it which is basically a two

level compartment where we have a liquid
compartment separated by a track and

membrane where basically to spores of this
membranes are about ten micro meters big

and then in this lower compartment
here for instance we have an A.T.P.

solution and a buffer in this upper
compartment so A.T.P. is going to slowly.

If you through the spores.

So we're going to create
basically localized events

where A.T.P.'s present at the surface and

then we'll see if I electrode scans in the
near field across this porous membrane.

Where do we see this negative
signal does Farraday current drop

when we read off the pores where data
piece produced and we can see that very

nicely These are the approach curves again
not going to go into details on that but

just to show you that this is
a quantitative methods of the Farraday

current you measure directly correlates
with the concentration of the analyze and

we see very nicely that
if we do it to them.

Ancient The image here at a distance
of about five marker meters

away from that surface we can image
these ports very very nicely.

Keep in mind the scale here is
actually in Pico AMSA we're

measuring the ferret they incur and
of hydrogen peroxide.

Again right as the signal drop
over here to signal drops.

Because eighty piece diffusing
through this porous and

gets into this quantitative
competitive reaction.

The glucose oxidase and the hex all kinds.

So this works beautifully and

in fact we have a program which aims at
using this technology in a true clinical

research environment at this corroded all
the surface that I've mentioned before.

Keep in mind again to corrode it body to
send a sample of about two three hundred

microns in diameter.

So our collaborators perfectly happy with
a twenty five micron electrode it doesn't

need to be any smaller than that to
position it actually here above to

scroll to body trigger actually by either
about potassium the polarization to

simulate vents trigger release
events at this corroded body and

measure what's happening with the A.T.P.
Khans a concentration in close vicinity.

I'll just give you a couple of very brief
example series here we see a superfood

corroded part of preparation which
actually read corroded bodies and

here we see the passenger position
actually a Bafta corroded body and

we see here the raw signal but

if we draw if I draw your attention
down here we have here actually.

Unfortunately it's a little small
printed here we hear here directly

the concentration of A.T.P. and
we see if we triggered a cell actually or

the cell ensemble we see that eighty piece
released in the shop spike then levels

out then we go back actually to a solution
where we don't have oxygen tension.

We go back to zero then we can trigger
a cell again A.T.P.'s released and so

on and so forth.

So we can do that over and over again.

In fact again I'm not going to bother you
with all the data we have cause to one

calibration in C two with
a known concentration of A.T.P.

before the cell experiments and run
afterwards to make sure that our sensor

is not drifting but quantitatively
measures A.T.P. at this face.

Of course we presented that to an agent
we were hoping to get funding for

that and then a reviewer who was
a very good reviewer I have to say

actually dig through the literature and
say that.

Six.
Hundred fifty million volts and

not only going to oxidize
how to turn proc side.

Actually the Cata colon means are also
oxidized at the same potential.

So how are you going to correct for that.

And in fact he was right.

We're going to have a signal that's going
to be convoluted by contributions from

Quetta colons infected was a great such
action and we ultimately got the project

funded because it turns out that we
can actually implement a second sensor

which measures the Quetta colon means at
the same time and corrects our eighty P.C.

well and this is actually shown here.

Now we have here destroy electro
the sample e where one electrode is

modified with ions imes measuring A.T.P.

the second electrode measures
to create a colon means and

we see that we have it primate background
signal here off the Quetta colon means.

And now we can correct
our eight to pieces.

For the additional contributions of
the catacombs and get the true A.T.P.

concentration which has been validated
with all kinds of essays and

with the literature bettas OK so
there's an example that in principle

these small biosensors can work
at Lafe biological interfaces and

provide us with molecular
level information either

with spatial resolution but also with
the temporal resolution that's required.

OK.

Why do we want to get smaller.

We want to get smaller because the second
project we have working on this if you

feel cells there we're trying to prove the
theory that the See if you are channel.

RIESS or basically triggers in exercise
told to release of A.T.P. and these

events happen that I mention a scale of
about hundred to two hundred nanometers So

we need an electrode which is small enough
to resolve such an event at the surface

and at the same time measure against this
one species that we're truly interested in

in this case A.T.P. So what we need to do
is we need to reduce that I mentioned so

I electrode to about two
hundred nanometers but

what comes along with that it means
that we have to be about a hundred

to two hundred on a metres away
from a face to do a quantitative.

Measurement.

Rest assured if you try to
position a two hundred nanometer

electrodes only by the current
in the see approach

to get to two hundred nanometers away from
the surface takes about a week to do that

if you're lucky you can do one
experiment and do this measurement.

So we had to find a different approach.

And that's again it's similar to systems
biology we have to integrate multiple

disciplines to get to an end a little tool
that allows us to do this measurement

at such a small scale and in fact
what we took advantage of is defect

that there is a whole range of analytical
technologies which work in the near field

of samples inherently And those are the
so-called scanning probe technologies so

when we take a very very small
tip which can be contacted.

For insulating and we talk about atomic
force microscope we are scanning tunneling

microscope the where we probe surfaces and

the topology of surfaces at
very very small dimensions.

So we said fine let's do the following.

Let's use one of these technologies
as a very expensive spacer for

a very small Elektra with that we would
like to deliberately position at our

biological surface.

So the idea was to take a technology
like atomic force my cross copy and

use this if empty place a vehicle to
position it two hundred nine a metre

Elektra look at it deliberately select
a distance of this biological surface

in micro fabrication has enabled
us to develop such a technology.

Again I'm going to be very brief on that
but I'm happy to give you more details in

the discussion of the words what we can
do with microfiber cation is we can build

down to about one hundred nano meter
Elektra What's that a position

to recessed from the insulating
tip of these air fam to appear and

that means actually since we micro
fabricate it we can make the spacer to see

if empty to correspond to the optimum
working distance for it is Elektra.

How are we doing that very briefly
we take a silicon nitride if

emptive we can coat any Elektra with
material you want onto the surface of.

Steve and then we're going to
insulate this entire theater.

And then we use a technology
called focused on be milling which

is available at Georgia Tech since
a couple of years to actually expose it

time to time electrodes here at the end of
the F.M.T. that basically cross-section

in through these electrodes layer and then
we reshape in the F.M.T. where the length

of this tiny tape now correlates to
the time meter of these electrodes.

So what happens now is
this is a half empty.

It is a spacer for this tiny electrode and
with the convoluted to current signal we

measure it is a electrode surface from
Actually it's distance dependence because

it's always going to move at a constant
distance across the apology of our surface

we can do that in the variety
of ways shapes and forms.

You can i just like to draw your attention
to this focused on beam Center which is

right now migrating into the M.R.C.
into the microelectronics

Research Center in about two weeks
it will be up and running again.

So everyone on campus has this tool
available to actually favorite Kate

at a scale of about or
with the precision of about

twenty nanometers without any
optical authority any mask and

you can literally fabricate basically
the electrodes inside these cantilevers.

So the main advantage of this
technology is now that we have much

much smaller electrodes and
we can very precisely position them but

we can actually take the very
same pile of chemistry and

image immobilised the now into these tiny
Elektra old sitting in this tape and

do exactly the same
measurement we did before but

two orders of magnitude smaller scale
in terms of climate change and.

In fact you're not limited to what
you can immobilize to how you can

modify this electrode surface.

You can put down to spiral
recognition elements like an amp

parametric enzymatic sensor.

We've had a project where we modified
the surface with a thin Mercury layer

forming a thin film career
electrode in the tip.

So you can do stripping well to
metry in an effective actually and

position it with the precision
of any effective.

You can for
instance use as an electrode layer.

Radio and if your exit eyes are read
I'm at the surface it's ph sensitive.

So you can put actually a PH
an imaging ph sensor in T.F.M.

tip and
look at PH distributions at the surface.

OK.

Again I'm not going to go
into details on that but

just to demonstrate to you
that this is actually working.

So in a first step.

We actually mobilized glucose
oxidase at this Alec Charles and

basically build an imaging glucose
pile sensor into the F.M.T.

we again you start to
compartment system here but

now the pores are only two
hundred nano meters in diameter.

OK And we'll see if we measure the glucose
by converting it with the glucose oxidase

immobilised at the tip at the Elektra we
see we see an enhanced current because

we're producing hydrogen peroxide and we
can very easily resolve with our image in

power sensor poorest that are about
two hundred nanometers in diameter and

now that corresponds about to the
dimensions of an exercise Toltec event.

We've done the same thing actually for

this project on cystic fibrosis
immobilizing sensor onto the C.F.M.

tape and this is work in progress
right now these are F. M.

images of the six lung at the seal cells
that were investigating together with

Emory University in effect what
we'd like to do now is image

that distribution of A.T.P.
with the letter a resolution of

about hundred nanometers across the
surface of the separates the little cells.

I can show your images yet
all of the A.T.P.

distributions were very close to that but
what I can tell you is that our A.T.P.

past sensor integrated into the if
MT is already working quantitatively

at the surface of this biological
systems and I hope in a couple of months

I'll be able to update you with actually
imaging experiments at the surface.

So in the last ten minutes.

I would like to expand a little bit and

give you sort of a future
outlook because now I've shown

to you that we can do to feed and
imaging electrochemistry at the same time.

Question is is that all we can
do in systems and a little X.

in the answer is No there's a lot of other
analytical techniques that we actually can

coing to create with this device platform
and I'm going to show you a couple

of them in a little more detail actually
very recently we've been able to combine

to scan improper technology
with infrared spectroscopy So

what they had allows us to do now
is to record infrared spectrum for

instance of whole cells of tissues
of membranes while we do have

a resolution to apocryphally on
top of this biological systems and

I'll show you in a minute actually how
that works OK another technology we're

working on is to make this electrodes even
smaller than they are already with like to

get the low hundred nanometers in
dimensions for these electrodes that

is a tricky fabrication problem that we're
working on now since a couple of years but

we already managed to make actually disk
Elektra lots by drilling holes through

the C.F. aim cantilevers that we fill
with the electrodes material and

then expose a tiny disk
Elektra's here at the end and

there we can get down to the maintenance
of about one hundred hundred twenty

nanometer us right now and

we hope we can further reduce that
below the hundred nanometer domain.

I think an exciting project that we just
started and we had a fantastic talk here

a couple of weeks ago from the University
of Washington on mass spectrometry and

imaging mass spectrometry that's something
that interests us since a number of years

in fact seven years ago we proposed
that in Siri we could use an E F M

to guide mass spectrometry
locally at biological surfaces

in fact that time when we proposed
that we had no clue how to make that.

But I'll show you.

Today a quick example that in fact we can
make an airframe tip which has a tiny hole

going through the C.F.M. tip and the large
metal Elektra the comically shaped

electrode on top which should allow you to
probe material off the surface of a life

biological specimen and
injected into a mass spectrometer.

So what that would mean
is that you can do a F.M.

guided mass spectrometry you always know
where off the power logical surface.

You're actually probing
the molecular material.

OK So just to give you
a quick example on the I.R.

spectroscopy if the idea is to use
actually a wave guide where the seven S.

and Taylor evanescent field protrudes
from the surface of the wave.

And that's where a logical
specimen is going to be present.

So in the first step.

We'll be able or we were aiming at
doing bulk infrared spectroscopy

while we probe actually the surface
with her resolution to feed

Of course we haven't been able to
do the biological systems yet but

we did it at a model system where
we demonstrated that I aspect of B.

and atomic force microscope we can
actually work at the same platform.

So what we did is we took actually a very
small attenuated total reflection infrared

waveguide couple infrared
radiation into the air and

produced a spot of about two
hundred fifty micro meters

of an evanescent field which produced
in the adjacent medium that means at

the surface of this way if God we can do
infrared spectroscopy and then the F.M.

was positioned right above this hot spot
to do two apocryphally at the same time.

Again we haven't been able to do that.

Logical system see it but
what we did is we deposit it actually

a crystal material at
the surface of this wave.

Which we slowly dissolve
the way from the surface and

recorded our infrared spectrum while
we recorded the topographical changes.

So in fact what we did is we built a tiny
infrared access point into the bottom

trough plate of an atomic force microscope
and what you see here is actually a Euro

feature Crystal in features that are
recorded with atomic force Macross copy

on top here as they slowly it is also a
way to see these features get smaller and

smaller and at the same time we see that
the absorption of your ear decreases and

decreases because we're going to
dissolve your your way out of this.

So-called evanescent field
which sticks up about

two micro meters from this we've got
surface into this it Jason medium.

And I can imagine.

Instead of having your your ear
precipitate up here we can have a cell or

a cell a sample or tissue sitting here and
we're going to probe the I.R.

spectral characteristics off
their tissue as it undergoes for

instance molecular changes or chemical
changes during similar processes but

at the same time we can
use our effective or

in future of if electrochemistry tape on
pulp of the very same biological specimen.

So now we're doing our aspect imaging
electrochemistry and to park a fee.

At the same time at the same sample
in a time and space correlated.

So that just as a quick
example on what's possible.

We're working on making that a lot more
sensitive this example here was by

no means sensitive enough we're creating
this tiny spot of infrared radiation and

trying to probe this molecular domain.

We have currently developments in the in
process and again we're probably not going

to be able to discuss all those to
use actually much more sensitive

infrared spectroscopy to do the same
measurement one way to do that is to

replace your standard brought painting for
a light source with a laser light source.

Fortunately lasers in the infrared
have also changed dramatically

over the past couple of years.

And let me keep to the next slide.

So the laser light sources we have not
is available in infrared spectroscopy

rather than the bulky large gas lasers
like C O two laser socio lasers.

It is heft as one slide to fast we
Noddy's have what is called Quantum

cascade leaves us which a tiny Samy come
back to lasers which to meet meet infrared

radiation anywhere between three and
twenty micro meters but at the same form

factor like to screen laser diode here
in this point or or the Red Laser dials.

That means we can do
infrared spectroscopy.

With a tiny tiny laser diode which
produces a very high energy density

in a very narrow spectrum regime and

we hope with that we'll get actually
the sensitivity or towards the sensitivity

that we are interested in to combine
this with atomic force microscope

is another project I'd like to bring close
to you that we are not limited actually to

using infrared spectroscopy we can even
go further in the wavelength regime and

move towards what is called
the far infrared actually or

nowadays more fancy called the terahertz
regime and when I had it in for it.

Spectroscopy in the early eighty's it was
called far infrared spectroscopy So I'd

like to stick to that term so that means
we're moving here from this vibrational

regime to actually a very very long way
of length up to about a thousand microns.

So the question is what are we.

Probing if we illuminate biological
material with a frequency

regime between thirty and
a thousand microns.

It turns out that even in the late sixty's
it was already predicted it actually

biological systems and in particular large
molecules should have actually longer

to do in the electrical modes that can be
excited at this very very long way flick.

So we're not going to see sharp absorption
features like we have Indian for it but

we're going to see very very broad
changes the question is are these

changes characteristic for an event
that relates to these molecules and

there was a landmark paper published in
two thousand in Applied Physics Letters

where it was demonstrated that this
long way flanks and in fact for

instance if D.N.A. hybridise is we see
a shift of the dielectric function.

So we don't see a sharp peak that we
can evaluate but we see basically

every franked if index shift of the
material at this very very long way from.

What's the puti of this technology.

The beauty is that this
detection of D.N.A.

hybrids is sation here is
stand totally label free.

We don't need to label D.N.A. anymore.

We're going to look basically at
this large dielectric function.

So that's a project we are currently
working on we're trying actually to make

a scanning probe tip which
can do top Hawker fifty and

near field turrets imaging at
the same time integrated into one.

And I'll just show you basically how this
tip looks like this is the monolithic

integration of this tip and in fact we
have now if first prototype of such

an effective which is surrounded
by a near field aperture

where we can extract terahertz
radiation and illuminate our sample.

So with that I'll just
highlight the other areas.

I'll promise to talk a little bit about or
if you're just a glimpse of what mass

spectroscopy or imaging mass spectroscopy
can look like you see this here on

the right hand side in fact this is
the first prototype of a tip that we made.

So it looks like an A.F.M.
tip with two pyramids.

But in fact there is a channel going
through the silicon nitride permit here.

This is a platinum cone on top and

this channel on the inside
is also lined with platinum.

So this actually looks like and
also that you use in your mastic

trauma to inject samples actually
into you for instance your ion tread.

OK So in fact this tip still can act
as an E.F. empted so you can position

basically a mass spec nozzle wherever
you wanted their biological surface and

then probe molecules off into
your mass back traumatic.

If we could do that we would have received
the funding already what I don't show you

here is of course how we're going to get
the molecules and particularly this very

small amount of molecules quantitatively
into mass spectrometer that's exactly

what we want to work on the next couple
of years how to probe off this tip and

get directly into the mass spectrometer
at the end in a quantitative fashion.

We've also demonstrated already it at
this type of tip technology can be

compliant for instance with
comm focal fluorescent imaging.

So we can take even a standard tool
to the cellular physiologist and

provide them with additional
information on top.

And electrochemistry.

By this combination.

And we've also shown in principle that we
can integrate more than one Elektra out

into the state and

do actually two independent electron
a little measurements at the same time.

So with their the hope that I've provided
you with a slightly different picture to

it and a little chemist hair Fuentes
think about biological entities and what

analytical chemistry can contribute to
eluding processes at biological entities.

We really would like to detect molecules
in a localized fashion at life

by a logical interfaces with the main
emphasis on and we think that's really

important to have corporative beauty and
synchronicity between the different

analytical techniques since our biological
systems are continuously changing we would

like to probe multiple parameters similar
tain Asli and we would like to do this

probing in what we would call an imaging
mold we would like to be able regardless

whether it's mass spectroscopy optical
spectroscopy electrochemistry apology.

We would like to be able to get this
information with spatial resolution.

So really the goal is to try and
complement systems biology with what we

call a systems and a little approach I've
put down here this was actually from

another presentation where the discussion
was always what is nano medicine so

there's so many expressions nowadays
around so we said well maybe if

you take to get a Systems Biology and
what we're doing in systems analytics

that might lead as a path to what we
might call in future and on a medicine.

So I want to end with a fairly
bold statement saying

that we can integrate disciplines
are very similar to systems biology

to actually develop
the complementary toolset

to probe this biological process
is in this concerted fission.

Was there like to come to an end thinker
the research group in particular.

Dr Christine Crohn's who is doing all
the amount of functional scanning

proclivity Look man all current and past.

Postdocs it's actually in
all the entirely up to date

on that the fabulous work was
Johns Hopkins on the curl

didn't bother his not assistant
professor at the University of Montreal

post-doc in our group does all the micro
favor cation young circle moon

in the group working also on
the multi-functional scanning probes.

Working on the if it feels cells and

most of the other group members working on
the complimentary in for its spectroscopy.

I think you for your attention.

I'm happy to answer any.